60 results on '"Cunha G"'
Search Results
2. Healthcare cost expenditure for robotic versus laparoscopic liver resection: a bottom-up economic evaluation.
- Author
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Pilz da Cunha G, Coupé VMH, Zonderhuis BM, Bonjer HJ, Erdmann JI, Kazemier G, Besselink MG, and Swijnenburg RJ
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- Humans, Male, Retrospective Studies, Female, Middle Aged, Aged, Hospital Costs, Cost-Benefit Analysis, Operative Time, Health Care Costs, Treatment Outcome, Time Factors, Laparoscopy economics, Hepatectomy economics, Robotic Surgical Procedures economics, Health Expenditures
- Abstract
Background: Minimally invasive liver surgery (MILS) is increasingly performed via the robot-assisted approach but may be associated with increased costs. This study is a post-hoc comparison of healthcare cost expenditure for robotic liver resection (RLR) and laparoscopic liver resection (LLR) in a high-volume center., Methods: In-hospital and 30-day postoperative healthcare costs were calculated per patient in a retrospective series (October 2015-December 2022)., Results: Overall, 298 patients were included (143 RLR and 155 LLR). Benefits of RLR were lower conversion rate (2.8% vs 12.3%, p = 0.002), shorter operating time (167 min vs 198 min, p = 0.044), and less blood loss (50 mL vs 200 mL, p < 0.001). Total per-procedure costs of RLR (€10260) and LLR (€9931) were not significantly different (mean difference €329 [95% bootstrapped confidence interval (BCI) €-1179-€2120]). Lower costs with RLR due to shorter surgical and operating room time were offset by higher disposable instrumentation costs resulting in comparable intraoperative costs (€5559 vs €5247, mean difference €312 [95% BCI €-25-€648]). Postoperative costs were similar for RLR (€4701) and LLR (€4684), mean difference €17 [95% BCI €-1357-€1727]. When also considering purchase and maintenance costs, RLR resulted in higher total per-procedure costs., Discussion: In a high-volume center, RLR can have similar per-procedure cost expenditure as LLR when disregarding capital investment., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)
- Published
- 2024
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3. Ovotesticular cords and ovotesticular follicles: New markers in a model of human mixed ovotestis.
- Author
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Baskin L, Cao M, Askel S, Li Y, and Cunha G
- Subjects
- Humans, Gonads, Ovotesticular Disorders of Sex Development
- Published
- 2024
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4. Does moderate hyperkalemia influence survival in HF? Insights from the MECKI score data base.
- Author
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Toto F, Salvioni E, Magrì D, Sciomer S, Piepoli M, Badagliacca R, Galotta A, Baracchini N, Paolillo S, Corrà U, Raimondo R, Lagioia R, Filardi PP, Iorio A, Senni M, Correale M, Cicoira M, Perna E, Metra M, Guazzi M, Limongelli G, Sinagra G, Parati G, Cattadori G, Bandera F, Bussotti M, Mapelli M, Cipriani M, Bonomi A, Cunha G, Re F, Vignati C, Garascia A, Lombardi C, Scardovi AB, Passantino A, Emdin M, Passino C, Santolamazza C, Girola D, Zaffalon D, Vizza D, De Martino F, and Agostoni P
- Subjects
- Humans, Retrospective Studies, Stroke Volume, Renin-Angiotensin System, Potassium, Heart Failure, Hyperkalemia diagnosis, Hyperkalemia epidemiology
- Abstract
Background: The prognostic role of moderate hyperkalemia in reduced ejection fraction (HFrEF) patients is still controversial. Despite this, it affects the use of renin-angiotensin-aldosterone system inhibitors (RAASi) with therapy down-titration or discontinuation., Objectives: Aim of the study was to assess the prognostic impact of moderate hyperkalemia in chronic HFrEF optimally treated patients., Methods and Results: We retrospectively analyzed MECKI (Metabolic Exercise test data combined with Cardiac and Kidney Indexes) database, with median follow-up of 4.2 [IQR 1.9-7.5] years. Data on K
+ levels were available in 7087 cases. Patients with K+ plasma level ≥ 5.6 mEq/L and < 4 mEq/L were excluded. Remaining patients were categorized into normal >4 and < 5 mEq/L (n = 4826, 68%) and moderately high ≥5.0 and ≤ 5.5 mEq/L (n = 496, 7%) K+ . Then patients were matched by propensity score in 484 couplets of patients. MECKI score value was 7% [IQR 3.1-14.1%] and 7.3% [IQR 3.4-15%] (p = 0.678) in patients with normal and moderately high K+ values while cardiovascular mortality events at two years follow-up were 41 (4.2%) and 33 (3.4%) (p = 0.333) in each group respectively., Conclusions: Moderate hyperkalemia does not influence patients' outcome in a large cohort of ambulatory HFrEF patients., Competing Interests: Declaration of Competing Interest None to declare., (Copyright © 2022 Elsevier B.V. All rights reserved.)- Published
- 2023
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5. A model to study human ovotesticular syndrome.
- Author
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Baskin L, Cao M, Derpinghaus A, Aksel S, Overland M, Li Y, and Cunha G
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- Female, Animals, Mice, Humans, Male, Mice, Nude, In Situ Hybridization, Fluorescence, Gonads, Ovary, Testis, Ovotesticular Disorders of Sex Development diagnosis, Ovotesticular Disorders of Sex Development metabolism, Ovotesticular Disorders of Sex Development pathology
- Abstract
Ovotesticular syndrome is a rare disorder of sex development characterized by the presence of testicular and ovarian tissue. The histologic characteristics of human testicular tissue are well defined by the presence of seminiferous cords or tubules containing TSPY-positive germ cells and Sox9-positive Sertoli cells surrounded by interstitial tissue containing cytochrome P450-positive Leydig cells and smooth muscle α-actin-positive peritubular myoid cells. The histological characteristics of the ovary can be defined by germ cell nests and the development of follicles. In contrast to the testis, the ovary has a paucity of defined specific protein markers, with the granulosa cell marker FOXL2 being the most widely used. In practice, defining the ovarian component of the ovotestis can be quite difficult. We developed a model of human ovotesticular syndrome by combining fetal human testis and ovary in a xenograft model. Ovotesticular xenografts were grown under the renal capsules of gonadectomized athymic nude mice for 6-32 weeks along with age matched control grafts of fetal testis and ovary. Forty ovotesticular xenografts and their controls were analyzed by histology, immunohistochemistry, and fluorescent in situ hybridization to determine the protein expression and karyotype of the cells within the grafts. The ovotesticular xenografts exhibited recognizable testicular and ovarian tissue based on testis-specific and ovary-specific markers defined above. The xenografts simulated a bipolar ovotestis in which the testicular and ovarian elements retain their separate histological characteristics and are separated by a well-defined border. This contrasts with the compartmentalized ovotestis previously described in the literature where the testicular tissue is surrounded by ovarian tissue or a mixed histology where testicular and ovarian tissues are interspersed throughout the gonad. In conclusion, we have characterized a human model of ovotestis which will allow a deeper understanding of ovotestis development in humans and facilitate a more accurate diagnosis of the ovotesticular syndrome., (Copyright © 2021. Published by Elsevier B.V.)
- Published
- 2023
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6. Response to: "Re ChEVAR from Below".
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Mees BME, Pilz da Cunha G, Lemmens CC, and Schurink GWH
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- Blood Vessel Prosthesis, Humans, Stents, Aortic Aneurysm, Abdominal surgery, Blood Vessel Prosthesis Implantation, Endovascular Procedures
- Published
- 2021
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7. Chimney Endovascular Aneurysm Repair from Below.
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Pilz da Cunha G, Lemmens CC, and Mees BME
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- Aged, Angioplasty, Balloon instrumentation, Aorta, Abdominal diagnostic imaging, Aorta, Abdominal surgery, Computed Tomography Angiography, Endoleak, Femoral Artery diagnostic imaging, Femoral Artery surgery, Humans, Male, Stents, Treatment Outcome, Ultrasonography, Interventional, Angioplasty, Balloon methods, Aortic Aneurysm, Abdominal surgery
- Published
- 2020
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8. Imaging the developing human external and internal urogenital organs with light sheet fluorescence microscopy.
- Author
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Isaacson D, McCreedy D, Calvert M, Shen J, Sinclair A, Cao M, Li Y, McDevitt T, Cunha G, and Baskin L
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- Humans, Image Processing, Computer-Assisted methods, Imaging, Three-Dimensional methods, Microscopy, Fluorescence methods, Specimen Handling methods, Urogenital System cytology
- Abstract
Technological advances in three-dimensional (3D) reconstruction techniques have previously enabled paradigm shifts in our understanding of human embryonic and fetal development. Light sheet fluorescence microscopy (LSFM) is a recently-developed technique that uses thin planes of light to optically section whole-mount cleared and immunolabeled biologic specimens. The advent of commercially-available light sheet microscopes has facilitated a new generation of research into protein localization and tissue dynamics at extremely high resolution. Our group has applied LSFM to study developing human fetal external genitalia, internal genitalia and kidneys. This review describes LSFM and presents our group's technique for preparing, clearing, immunostaining and imaging human fetal urogenital specimens. We then present light sheet images and videos of each element of the developing human urogenital system. To the extent of our knowledge, the work conducted by our laboratory represents the first description of a method for performing LSFM on the full human urogenital system during the embryonic and fetal periods., (Copyright © 2019. Published by Elsevier B.V.)
- Published
- 2020
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9. Presence of parasite DNA in clinically unaffected nasal mucosa during cutaneous leishmaniasis caused by Leishmania (Viannia) braziliensis.
- Author
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Canário A, Queiroz M, Cunha G, Cavalcante T, Riesz V, Sharma R, de Noronha A, Correia T, Barral-Netto M, Barral A, Khouri R, and Boaventura V
- Subjects
- Adult, Brazil, Cross-Sectional Studies, Female, Humans, Male, Middle Aged, Tropism physiology, Young Adult, DNA, Protozoan genetics, DNA, Protozoan isolation & purification, Leishmania braziliensis genetics, Leishmaniasis, Cutaneous parasitology, Nasal Mucosa chemistry
- Abstract
Objectives: We aimed to detect Leishmania DNA carriage in nasal mucosa of individuals with cutaneous leishmaniasis (CL) caused by Leishmania (Viannia) braziliensis., Methods: A cross-sectional study was performed in all individuals with CL without nasal lesions (n = 153) attended within 2 years in an endemic area of L. (Viannia) braziliensis in Bahia (Brazil). An otorhinolaryngologist assessed the clinical status of the nasal mucosa by anterior rhinoscopy and endoscopic examinations. Swab samples were collected for parasite DNA detection by PCR from all individuals before standard treatment for leishmaniasis. A second evaluation 3 months after treatment was performed to assess clinical outcomes., Results: Parasite DNA was detected in 7.8% (12/153) of clinically healthy nasal mucosa of individuals with CL. Interestingly, DNA was more frequently identified in individuals with more skin lesions (median 1.5, interquartile range (IQR) 1-3.5 versus 1.0, IQR 1-1.5; p 0.044), or larger injuries (median 2.7, IQR 2-3.8 versus 1.6, IQR 1-2.5; p 0.013). Additionally, the disease of those individuals with positive PCR evolved more frequently to unusual forms of leishmaniasis (recidiva cutis and disseminated) (45.5% (5/11) versus 11.5% (14/122); p 0.009), and required more cycles of treatment to reach clinical cure (median 2, IQR 1-4 versus 1, IQR 1-2; p 0.05)., Conclusion: These findings suggest an early parasite tropism to nasal mucosa in L. (Viannia) braziliensis infection and a clinical phenotype of CL cases associated with parasite DNA in nasal mucosa. Future studies should evaluate whether PCR of nasal swab samples could serve as a prognostic tool for individuals at risk of mucocutaneous leishmaniasis., (Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.)
- Published
- 2019
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10. Advanced MRI study of migrainous infarction presenting as cortical laminar necrosis - Case report and literature review.
- Author
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Morais R, Sobral F, Cunha G, Brito O, and Santana I
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- Adult, Brain pathology, Diagnosis, Differential, Female, Humans, Infarction diagnosis, Migraine Disorders diagnosis, Infarction pathology, Magnetic Resonance Imaging, Migraine Disorders pathology, Necrosis pathology
- Published
- 2018
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11. Lightsheet fluorescence microscopy of branching human fetal kidney.
- Author
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Isaacson D, Shen J, McCreedy D, Calvert M, McDevitt T, Cunha G, and Baskin L
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- Antigens, CD immunology, Cadherins immunology, Gestational Age, Homeodomain Proteins immunology, Humans, Kidney immunology, Morphogenesis, Ureter immunology, Fluorescent Antibody Technique, Direct, Kidney embryology, Microscopy, Fluorescence methods, Ureter embryology
- Published
- 2018
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12. Renal Subcapsular xenografing of human fetal external genital tissue - A new model for investigating urethral development.
- Author
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Isaacson D, Shen J, Cao M, Sinclair A, Yue X, Cunha G, and Baskin L
- Subjects
- Animals, Endocrine Disruptors pharmacology, Female, Genitalia, Female growth & development, Humans, Hypospadias drug therapy, Kidney embryology, Male, Mice, Receptors, Androgen metabolism, Hypospadias pathology, Organogenesis physiology, Penis embryology, Urethra drug effects
- Abstract
In this paper, we introduce our novel renal subcapsular xenograft model for the study of human penile urethral and clitoral development. We grafted fifteen intact fetal penes and clitorides 8-11 weeks fetal age under the renal capsules of gonadectomized athymic mice. The mice were treated with a subcutaneous pellet of dihydrotestosterone (DHT), diethylstilbestrol (DES) or untreated with hormones. Xenografts were harvested after fourteen days of growth and analyzed via serial histologic sectioning and immunostaining for Ki-67, cytokeratins 6, 7 and 10, uroplakin and the androgen receptor. Non-grafted specimens of similar fetal age were sectioned and immunostained for the same antigenic markers. 14/15 (93.3%) grafts were successfully propagated and harvested. The developing urethral plate, urethral groove, tubular urethra, corporal bodies and preputial lamina were easily identifiable. These structures demonstrated robust cellularity, appropriate architecture and abundant Ki-67 expression. Expression patterns of cytokeratins 6, 7 and 10, uroplakin and the androgen receptor in xenografted specimens demonstrated characteristic male/female differences analogous to non-grafted specimens. DHT treatment reliably produced tubularization of nascent urethral and vestibular structures and male patterns of androgen receptor expression in grafts of both genetic sexes while estrogenic or hormonally absent conditions reliably resulted in a persistent open urethral/vestibular groove and female patterns of androgen receptor expression. This model's success enables further study into causal pathways by which endocrine-disrupting and endocrine-mimicking substances may directly cause disruption of normal human urethral development or hypospadias., (Copyright © 2017 International Society of Differentiation. Published by Elsevier B.V. All rights reserved.)
- Published
- 2017
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13. Cortical control of vertical and horizontal saccades in progressive supranuclear palsy: An exploratory fMRI study.
- Author
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Lemos J, Pereira D, Almendra L, Rebelo D, Patrício M, Castelhano J, Cunha G, Januário C, Cunha L, Freire A, and Castelo-Branco M
- Subjects
- Aged, Aged, 80 and over, Brain Mapping, Cerebral Cortex diagnostic imaging, Cerebrovascular Circulation physiology, Eye Movement Measurements, Female, Humans, Magnetic Resonance Imaging, Male, Middle Aged, Neural Pathways diagnostic imaging, Neural Pathways physiopathology, Neuropsychological Tests, Oxygen blood, Supranuclear Palsy, Progressive diagnostic imaging, Cerebral Cortex physiopathology, Saccades physiology, Supranuclear Palsy, Progressive physiopathology
- Abstract
Progressive supranuclear palsy (PSP) is a neurodegenerative disorder showing predominant brainstem involvement, characterized by marked slowing of rapid eye movements (saccades), particularly along the vertical plane. While the contribution of the brainstem damage for the saccadic disturbance in PSP has been extensively studied, much less is known about its cortical and subcortical pathomechanisms. We measured reflexive (prosaccades) and voluntary (antisaccades) saccades in the vertical and horizontal plane in PSP patients (n=8) and controls (n=10) in an eye tracking study, followed by the measurement of blood oxygenation-level dependent (BOLD) activation (PSP, n=6; controls, n=10) during similar saccade paradigms. Behaviorally, PSP patients evidenced slower and lower amplitude prosaccades (horizontal and vertical) and lower amplitude antisaccades (vertical) than controls. Functionally, patients showed decreased frontostriatal BOLD activation during prosaccades (horizontal and vertical) and antisaccades (vertical), relative to controls. Additionally, PSP patients showed less default mode network (DMN) deactivation than controls for all types of saccades. Within groups, controls showed no BOLD differences between horizontal and vertical prosaccades while PSP patients demonstrated greater DMN deactivation during vertical prosaccades. Both groups evidenced greater DMN deactivation during vertical antisaccades when compared to their horizontal counterpart and patients further showed relative frontostriatal BOLD hypoactivity during vertical antisaccades. We found fMRI evidence of frontostriatal hypoactivity in PSP patients relative to controls, especially during vertical saccades. These new findings highlight the impact of cortical impairment in saccadic disturbance of PSP., (Copyright © 2016 Elsevier B.V. All rights reserved.)
- Published
- 2017
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14. Complex epithelial remodeling underlie the fusion event in early fetal development of the human penile urethra.
- Author
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Shen J, Overland M, Sinclair A, Cao M, Yue X, Cunha G, and Baskin L
- Subjects
- Clitoris growth & development, Clitoris ultrastructure, Epithelium growth & development, Female, Genitalia, Female growth & development, Genitalia, Female ultrastructure, Humans, Hypospadias physiopathology, Male, Microscopy, Electron, Scanning, Penis growth & development, Urethra growth & development, Epithelium ultrastructure, Fetal Development, Penis ultrastructure, Urethra ultrastructure
- Abstract
We recently described a two-step process of urethral plate canalization and urethral fold fusion to form the human penile urethra. Canalization ("opening zipper") opens the solid urethral plate into a groove, and fusion ("closing zipper") closes the urethral groove to form the penile urethra. We hypothesize that failure of canalization and/or fusion during human urethral formation can lead to hypospadias. Herein, we use scanning electron microscopy (SEM) and analysis of transverse serial sections to better characterize development of the human fetal penile urethra as contrasted to the development of the human fetal clitoris. Eighteen 7-13 week human fetal external genitalia specimens were analyzed by SEM, and fifteen additional human fetal specimens were sectioned for histologic analysis. SEM images demonstrate canalization of the urethral/vestibular plate in the developing male and female external genitalia, respectively, followed by proximal to distal fusion of the urethral folds in males only. The fusion process during penile development occurs sequentially in multiple layers and through the interlacing of epidermal "cords". Complex epithelial organization is also noted at the site of active canalization. The demarcation between the epidermis of the shaft and the glans becomes distinct during development, and the epithelial tag at the distal tip of the penile and clitoral glans regresses as development progresses. In summary, SEM analysis of human fetal specimens supports the two-zipper hypothesis of formation of the penile urethra. The opening zipper progresses from proximal to distal along the shaft of the penis and clitoris into the glans in identical fashion in both sexes. The closing zipper mechanism is active only in males and is not a single process but rather a series of layered fusion events, uniquely different from the simple fusion of two epithelial surfaces as occurs in formation of the palate and neural tube., (Copyright © 2016 International Society of Differentiation. Published by Elsevier B.V. All rights reserved.)
- Published
- 2016
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15. Methods for studying human organogenesis.
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Cunha G, Overland M, Li Y, Cao M, Shen J, Sinclair A, and Baskin L
- Subjects
- Animals, Ecotoxicology, Gene Expression Regulation, Developmental, Humans, Mice, Aborted Fetus transplantation, Cell Differentiation genetics, Organogenesis genetics
- Abstract
This review details methods for utilizing D & C suction abortus specimens as a source of human fetal organs to study the morphogenetic and molecular mechanisms of human fetal organ development. By this means it is possible to design experiments elucidating the molecular mechanisms of human fetal organ development and to compare and contrast human developmental mechanisms with that of laboratory animals. Finally human fetal organs can be grown in vivo as grafts to athymic mice, thus allowing ethical analysis of potential adverse effects of environmental toxicants., (Copyright © 2015 International Society of Differentiation. Published by Elsevier B.V. All rights reserved.)
- Published
- 2016
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16. Investigation of sexual dimorphisms through mouse models and hormone/hormone-disruptor treatments.
- Author
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Ipulan LA, Raga D, Suzuki K, Murashima A, Matsumaru D, Cunha G, and Yamada G
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- Animals, Disease Models, Animal, Female, Genitalia embryology, Hormone Antagonists administration & dosage, Integrases genetics, Male, Mice, Morphogenesis drug effects, Morphogenesis genetics, Organogenesis drug effects, Genitalia growth & development, Hormones metabolism, Organogenesis genetics, Sex Differentiation genetics
- Abstract
Sexual dimorphism in mouse reproductive tissues is observable in adult, post-natal, and embryonic stages. The development of sexually dimorphic tissues starts with an ambisexual structure. It is followed by sex-specific organogenesis as guided by different signaling pathways that occur from late embryonic stages. The measurement of the anogenital distance (AGD), and the observation of the external genitalia are practical ways to distinguish male and female pups at birth and thereafter. Careful observation of the morphological or histological features and the molecular signatures of the external genitalia and perineum enable identification of sex or feminization/masculinization of embryos. Aberrations in hormone signaling via castration or treatment with hormones or hormone disruptors result in dysmorphogenesis of reproductive tissues. Several hormone disruptors have been used to modulate different aspects of hormone action through competitive inhibition and exogenous hormone treatment. Concomitantly, the vast advancement of conditional mutant mouse analysis leads to the frequent utilization of Cre recombination technology in the study of reproductive/urogenital tissue development. Mouse Cre-lines that are tissue-specific and cell-specific are also effective tools in identifying the molecular mechanisms during sexually dimorphic development. Cre-lines applicable to different cell populations in the prostate, seminal vesicles, testis and ovaries, and mammary glands are currently being utilized. In the external genitalia and perineum, Cre lines that examine the signaling pathways of cells of endodermal, ectodermal, and mesenchymal origin reveal the roles of these tissues in the development of the external genitalia. The interaction of hormones and growth factors can be examined further through a variety of techniques available for researchers. Such cumulative information about various technologies is summarized., (Copyright © 2015 International Society of Differentiation. Published by Elsevier B.V. All rights reserved.)
- Published
- 2016
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17. The power and perils of animal models with urogenital anomalies: handle with care.
- Author
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Hutson JM, Baskin LS, Risbridger G, and Cunha GR
- Subjects
- Animals, Humans, Male, Species Specificity, Cryptorchidism pathology, Disease Models, Animal, Hypospadias pathology
- Abstract
Congenital abnormalities of the urogenital tracts form a major part of clinical practice for paediatric urologists, but their knowledge of normal and abnormal development is often limited. Advances in understanding frequently come from studying experimental findings from animal models, however, most clinicians underestimate both the power and perils of extrapolating scientific knowledge from animals. In this review, the key issues that urologists need to understand in order to link animal studies to clinical practice are discussed. Urologists must avoid the traps of anthropomorphism (assuming humans are always the same as animal models) or anthropocentrism (assuming humans are too different from animal models). This review used two common disorders: hypospadias and undescended testes., (Copyright © 2014. Published by Elsevier Ltd.)
- Published
- 2014
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18. Susceptibility profiles and correlation with pneumococcal serotypes soon after implementation of the 10-valent pneumococcal conjugate vaccine in Brazil.
- Author
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Mott M, Caierão J, Rosa da Cunha G, Rodrigues Perez LR, Matusiak R, Pilger de Oliveira KR, d'Azevedo PA, and Dias C
- Subjects
- Adolescent, Adult, Aged, Aged, 80 and over, Anti-Bacterial Agents therapeutic use, Brazil, Child, Child, Preschool, Clindamycin therapeutic use, Erythromycin therapeutic use, Humans, Infant, Levofloxacin therapeutic use, Macrolides therapeutic use, Microbial Sensitivity Tests, Middle Aged, Pneumococcal Infections drug therapy, Pneumococcal Infections prevention & control, Pneumococcal Vaccines administration & dosage, Serotyping, Streptococcus pneumoniae drug effects, Streptococcus pneumoniae isolation & purification, Tetracycline therapeutic use, Trimethoprim, Sulfamethoxazole Drug Combination therapeutic use, Young Adult, Drug Resistance, Multiple, Bacterial, Pneumococcal Infections microbiology, Streptococcus pneumoniae classification
- Abstract
Objectives: To evaluate the susceptibility patterns among Streptococcus pneumoniae recovered during the years 2010-2012 and to correlate these with serotypes., Methods: Pneumococci from invasive sites were serotyped by sequential multiplex PCR and/or Quellung reaction. Etest strips were used to determine the minimal inhibitory concentrations, and the Clinical and Laboratory Standards Institute (CLSI) guidelines were used for interpretation. Genetic determinants of macrolide resistance were assessed by PCR, and the occurrence of the D phenotype was analyzed following the recommendations of the CLSI., Results: One hundred fifty-nine S. pneumoniae were studied; most were recovered from blood and were associated with serotypes 14, 3, 4, 23F, 20, 7F, 12F, 19A, and 19F. Pneumococcal conjugate vaccine PCV7, PCV10, and PCV13 and 23-valent polysaccharide vaccine serotypes represented 38.2%, 48.7%, 64.5%, and 85.5%, respectively. β-Lactam non-susceptibility (non-meningitis) was basically related to serotype 19A. For meningitis, it was observed in 21.4% (serotypes 14, 3, 9V, 23F, and 24F). Resistance to erythromycin occurred in 8.2% and mefA was the most common macrolide genetic determinant. One isolate was resistant to levofloxacin. Non-susceptibility to trimethoprim-sulfamethoxazole was 37.7% and to tetracycline was 22.0%., Conclusions: Our population of pneumococci represents a transition era, soon after the introduction of PCV10. Non-susceptible patterns were found to be associated with classical PCV serotypes (especially serotype 14), which is still highly prevalent, and non-PCV10 ones (19A), which may disseminate, occupying the biological niche left by the vaccine serotypes., (Copyright © 2013 The Authors. Published by Elsevier Ltd.. All rights reserved.)
- Published
- 2014
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19. Colonization of Streptococcus mutans on esthetic brackets: self-ligating vs conventional.
- Author
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do Nascimento LE, Pithon MM, dos Santos RL, Freitas AO, Alviano DS, Nojima LI, Nojima MC, and Ruellas AC
- Subjects
- Adult, Analysis of Variance, Bacterial Adhesion, Colony Count, Microbial, Esthetics, Dental, Humans, Male, Orthodontic Appliance Design, Orthodontic Brackets microbiology, Streptococcus mutans physiology
- Abstract
Introduction: Self-ligating orthodontic brackets rely on clips, rather than ligatures, to hold the archwire in place. It is unknown whether replacing ligatures with clips affects the adherence of Streptococcus mutans. The aim of this research was to evaluate whether self-ligating brackets have an advantage over conventional brackets as determined by the adherence of S mutans., Methods: The sample consisted of 50 esthetic brackets, divided into 3 experimental groups and 2 control groups of 10 brackets each. Two experimental groups were active self-ligating brackets (QuicKlear; Forestadent, Pforzheim, Germany; and In-Ovation C; Dentsply GAC, Bohemia, NY); the other was a passive self-ligating bracket (Damon 3; Ormco, Glendora, Calif). The 2 control groups were conventional brackets (Mystique; Dentsply GAC; and Clarity; 3M Unitek, Monrovia, Calif). The brackets were randomly bonded to the canines, first and second premolars, and first and second molars in the mandibular left hemiarch of 10 male participants. Biofilm was collected from the tooth surfaces before bonding and from the brackets on day 21 and placed in Petri dishes containing Mitis salivarius agar. The brackets were removed on day 28 and examined by using scanning electron microscopy. Statistical analysis, analysis of variance, and the Tukey correction with a P value of 0.05 were used., Results: The greatest numbers of colonies were found in an active self-ligating bracket group (In-Ovation C), and the fewest colonies were in a conventional bracket group (Clarity). The largest colonies formed on active self-ligating brackets. In the slot, the greatest formation was in a control group (Mystique)., Conclusions: Self-ligating esthetic brackets do not promote greater or lesser S mutans colonization when compared with conventional brackets. Differences were found to be related to the material composition of the bracket., (Copyright © 2013 American Association of Orthodontists. Published by Mosby, Inc. All rights reserved.)
- Published
- 2013
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20. Specific morphogenetic events in mouse external genitalia sex differentiation are responsive/dependent upon androgens and/or estrogens.
- Author
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Rodriguez E Jr, Weiss DA, Ferretti M, Wang H, Menshenia J, Risbridger G, Handelsman D, Cunha G, and Baskin L
- Subjects
- Androgens pharmacology, Animals, Castration, Diethylstilbestrol pharmacology, Dihydrotestosterone pharmacology, Estrogens pharmacology, Estrogens, Non-Steroidal pharmacology, Female, Genitalia anatomy & histology, Male, Mice, Mice, Mutant Strains, Receptors, Androgen genetics, Androgens deficiency, Estrogens deficiency, Genitalia growth & development, Morphogenesis drug effects, Sex Differentiation drug effects, Sex Differentiation genetics
- Abstract
The objective of this study was to perform a comprehensive morphologic analysis of developing mouse external genitalia (ExG) and to determine specific sexual differentiation features that are responsive to androgens or estrogens. To eliminate sex steroid signaling postnatally, male and female mice were gonadectomized on the day of birth, and then injected intraperitoneally every other day with DES (200 ng/g), DHT (1 μg/g), or oil. On day-10 postnatal male and female ExG were dissected, fixed, embedded, serially sectioned and analyzed. We identified 10 sexually dimorphic anatomical features indicative of normal penile and clitoral differentiation in intact mice. Several (but not all) penile features were impaired or abolished as a result of neonatal castration. Those penile features remaining after neonatal castration were completely abolished with attendant clitoral development in androgen receptor (AR) mutant male mice (X(Tfm)/Y and X/Y AR-null) in which AR signaling is absent both pre- and postnatally. Administration of DHT to neonatally castrated males restored development of all 10 masculine features to almost normal levels. Neonatal ovariectomy of female mice had little effect on clitoral development, whereas treatment of ovariectomized female mice with DHT induced partial masculinization of the clitoris. Administration of DES to neonatally gonadectomized male and female mice elicited a spectrum of development abnormalities. These studies demonstrate that the presence or absence of androgen prenatally specifies penile versus clitoral identity. Differentiated penile features emerge postnatally and are sensitive to and dependent upon prenatal or pre- and postnatal androgen. Emergence of differentiated clitoral features occurs postnatally in either intact or ovariectomized females. It is likely that each penile and clitoral feature has a unique time-course of hormonal dependency/sensitivity., (Copyright © 2012 International Society of Differentiation. Published by Elsevier B.V. All rights reserved.)
- Published
- 2012
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21. A green strategy for desorption of trihalomethanes adsorbed by humin and reuse of the fixed bed column.
- Author
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Cunha GC, Romão LP, Santos MC, Costa AS, and Alexandre MR
- Subjects
- Adsorption, Spectroscopy, Fourier Transform Infrared, Environmental Restoration and Remediation methods, Humic Substances, Trihalomethanes chemistry
- Abstract
The objective of the present work was to develop a thermal desorption method for the removal of trihalomethanes (THM) adsorbed by humin, followed by multiple recycling of the fixed bed column in order to avoid excessive consumption of materials and reduce operating costs. The results obtained for adsorption on a fixed bed column confirmed the effectiveness of humin as an adsorbent, extracting between 45.9% and 90.1% of the total THM (TTHM). In none of the tests was the column fully saturated after 10h. Experiments involving thermal desorption were used to evaluate the potential of the technique for column regeneration. The adsorptive capacity of the humin bed increased significantly (p<0.05) between the first and fifth desorption cycle, by 18.9%, 18.1%, 24.2%, 20.2% and 24.2% for CHBr(3), CHBr(2)Cl, CHBrCl(2), CHCl(3) and TTHM, respectively., (Copyright © 2011 Elsevier B.V. All rights reserved.)
- Published
- 2012
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22. The individual and combined metabolite profiles (metabolomics) of dibutylphthalate and di(2-ethylhexyl)phthalate following a 28-day dietary exposure in rats.
- Author
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van Ravenzwaay B, Coelho-Palermo Cunha G, Strauss V, Wiemer J, Leibold E, Kamp H, Walk T, Mellert W, Looser R, Prokoudine A, Fabian E, Krennrich G, and Herold M
- Subjects
- Administration, Oral, Animals, Biomarkers blood, Chromatography, High Pressure Liquid, Dose-Response Relationship, Drug, Drug Synergism, Female, Gas Chromatography-Mass Spectrometry, Liver drug effects, Liver metabolism, Liver pathology, Male, Metabolomics methods, Organ Size drug effects, Rats, Rats, Wistar, Sex Factors, Tandem Mass Spectrometry, Toxicity Tests, Chronic, Dibutyl Phthalate toxicity, Diethylhexyl Phthalate toxicity, Environmental Pollutants toxicity, Metabolome drug effects
- Abstract
Metabolite profiles (metabolomics) of plasma samples of Wistar rats dosed with di(2-ethylhexyl)phthalate (DEHP - 3000ppm) and dibutylphthalate (DBP - 150, 1000 and 7000ppm) were individually determined in 28 days dietary studies. In addition, profiles of combined exposure to 3000ppm DEHP and either 150, 1000 or 7000ppm DBP were determined. High dose levels induced more profound metabolite changes in males than in females for both compounds. At 150ppm DBP (NOEL for toxicity) there were very few (
- Published
- 2010
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23. Smooth muscle differentiation and patterning in the urinary bladder.
- Author
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Tasian G, Cunha G, and Baskin L
- Subjects
- Hedgehog Proteins physiology, Humans, Body Patterning, Cell Differentiation, Muscle, Smooth embryology, Urinary Bladder embryology
- Abstract
Smooth muscle differentiation and patterning is a fundamental process in urinary bladder development that involves a complex array of local environmental factors, epithelial-mesenchymal interaction, and signaling pathways. An epithelial signal is necessary to induce smooth muscle differentiation in the adjacent bladder mesenchyme. The bladder epithelium (urothelium) also influences the spatial organization of the bladder wall. Sonic hedgehog (Shh), which is expressed by the urothelium, promotes mesenchymal proliferation and induces differentiation of smooth muscle from embryonic bladder mesenchyme. Shh, whose signal is mediated through various transcription factors including Gli2 and BMP4, is likely also important in the patterning of bladder smooth muscle. However, it is not known to what extent early mediators of mesenchymal migration, other Shh-associated transcription factors, and crosstalk between the Shh signaling cascade and other pathways are involved in the patterning of bladder smooth muscle. Here we review the role of epithelial-mesenchymal interaction and Shh signaling in smooth muscle differentiation and patterning in the bladder. We also discuss emerging signaling molecules, transcription factors, and mesenchyme properties that might be fruitful areas of future research in the process of smooth muscle formation in the bladder., (Copyright © 2010 International Society of Differentiation. Published by Elsevier B.V. All rights reserved.)
- Published
- 2010
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24. Urothelium-derived Sonic hedgehog promotes mesenchymal proliferation and induces bladder smooth muscle differentiation.
- Author
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Cao M, Tasian G, Wang MH, Liu B, Cunha G, and Baskin L
- Subjects
- Animals, Biomarkers metabolism, Female, Hedgehog Proteins genetics, Mice, Muscle, Smooth cytology, Pregnancy, Urothelium cytology, Cell Differentiation physiology, Cell Proliferation, Hedgehog Proteins metabolism, Mesoderm physiology, Muscle, Smooth physiology, Urinary Bladder anatomy & histology, Urinary Bladder physiology, Urothelium metabolism
- Abstract
Induction of smooth muscle differentiation from bladder mesenchyme depends on signals that originate from the urothelium. We hypothesize Sonic hedgehog (Shh) is the urothelial signal that promotes bladder mesenchymal proliferation and induces bladder smooth muscle differentiation. Pregnant FVB mice were euthanized on embryonic day (E) 12.5 and fetal bladders were harvested. Two experimental protocols were utilized: Specimens were sized by serial sectioning. Cell counts were performed after trypsin digestion. Immunohistochemistry was performed to detect smooth muscle-specific protein expression. alpha-Actin expression was quantified using Western blot. All specimens were viable at 72h. BLM cultured without Shh survived but did not grow or undergo smooth muscle differentiation. IB cultured without Shh and BLM cultured with Shh grew and expressed smooth muscle proteins at 72h. IB cultured with Shh were larger and contained more cells than IB cultured without Shh (all p<0.05). Increasing Shh concentration from 48 to 480nM did not change bladder size, cell counts, or the level of alpha-actin expression. Prior to culture, IB did not express alpha-actin. After culture of IB in Shh-deficient media, alpha-actin was detected throughout the mesenchyme except in the submucosal layer. The IB submucosa was thinner after culture with 48nM Shh and smooth muscle completely obliterated the submucosa after culture with 480nM Shh. In fetal mouse bladders, urothelium-derived Shh is necessary for mesenchymal proliferation and smooth muscle differentiation. Shh concentration affects mesenchymal proliferation and patterning of bladder smooth muscle.
- Published
- 2010
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25. Iron deficiency causes duodenum mucosal hyperplasia in male Wistar rats.
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Cunha GC, van Ravenzwaay B, Mellert W, and Kaufmann W
- Subjects
- Animals, Hyperplasia, Intestinal Mucosa pathology, Iron blood, Male, Rats, Rats, Wistar, Duodenum pathology, Iron Deficiencies
- Abstract
Administration of an iron-deficient diet to Wistar rats resulted within 14 days in reduced serum iron concentrations, a microcytic hypochromic anemia, characteristic for impaired hemoglobin synthesis, and an increase of duodenal epithelial cell proliferation. After 5 weeks of iron deficiency, hypochromic microcytic anemia and a clear increase of duodenum weight but no pronounced effects on cell proliferation was observed. Increased duodenum weights corresponded to significant increases in mucosal area, indicating a diffuse, simple mucosal hyperplasia. The sequence of events following iron depletion thus appears to be: (1) reduced serum iron levels, (2) induction of hypochromic microcytic anemia, (3) increased duodenal epithelial cell proliferation, and (4) increased duodenal weight (increased mucosal area). Iron deficiency anemia was rapidly reversible after a 2-week recovery period. However, increased duodenum weights were still noted at that time. Intramuscular iron supplementation in animals fed with iron-deficient diet maintained body iron levels not below normal values, and neither anemia nor increased duodenum cell proliferation were detected after 14 days. A 5-week iron supplementation period resulted in slightly increased serum iron values, and slightly decreased duodenal epithelial cell proliferation. Thus, increased duodenum mucosal hyperplasia was shown to be secondary to depletion of body iron and anemia and reflects an attempt to increase iron absorption to counteract iron deficiency.
- Published
- 2008
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26. Standardization of the perchlorate discharge assay for thyroid toxicity testing in rats.
- Author
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Coelho-Palermo Cunha G and van Ravenzwaay B
- Subjects
- Animals, Chlorodiphenyl (54% Chlorine) toxicity, Hyperplasia, Hypertrophy, Iodine Radioisotopes, Male, Phenobarbital toxicity, Potassium Iodide toxicity, Propylthiouracil toxicity, Rats, Rats, Wistar, Reproducibility of Results, Sensitivity and Specificity, Thyrotropin blood, Thyrotropin drug effects, Thyroxine blood, Thyroxine drug effects, Triiodothyronine blood, Triiodothyronine drug effects, beta-Naphthoflavone toxicity, Perchlorates, Potassium Compounds, Thyroid Function Tests standards, Thyroid Gland drug effects, Toxicity Tests standards
- Abstract
The perchlorate discharge assay (PDA) is potentially of high diagnostic value to distinguish between direct and indirect thyroid toxicity mechanisms, provided that standard treatment times are established and positive controls yield reproducible results. Therefore the PDA was evaluated after 2 and/or 4 weeks of treatment with positive control compounds in rats. Phenobarbital, Aroclor 1254 and beta-naphthoflavone (indirect toxic mechanism) enhanced thyroidal radioiodide accumulation, and the administration of potassium perchlorate had no effect on thyroid: blood (125)I ratio. Phenobarbital caused follicular cell hypertrophy and hyperplasia in the thyroid and centrilobular hypertrophy in the liver, without effects on serum triiodotyronine (T(3)), thyroxine (T(4)) levels. Thyroid-stimulating hormone (TSH) levels were moderately increased. Propylthiouracil (direct toxic mechanism) caused severe thyroid follicular cell hypertrophy and hyperplasia, reduced serum T(3) and T(4) levels and increased serum TSH levels, and reduced thyroidal radioiodide accumulation; perchlorate administration significantly reduced thyroid: blood (125)I ratio, demonstrating an iodide organification block. Potassium iodide (direct toxic mechanism) virtually blocked thyroidal radioiodide accumulation, without significant effects on serum T(3), T(4), and TSH levels and a microscopic correlate for higher thyroid weights. Thus, positive controls yielded reproducible results and we conclude that both the 2- and 4-week PDA is suitable to distinguish between direct and indirect thyroid toxicity mechanisms.
- Published
- 2007
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27. The use of metabolomics for the discovery of new biomarkers of effect.
- Author
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van Ravenzwaay B, Cunha GC, Leibold E, Looser R, Mellert W, Prokoudine A, Walk T, and Wiemer J
- Subjects
- 4-Hydroxyphenylpyruvate Dioxygenase antagonists & inhibitors, 4-Hydroxyphenylpyruvate Dioxygenase metabolism, Androgen Antagonists toxicity, Animals, Antithyroid Agents toxicity, Chromatography, Liquid, Dose-Response Relationship, Drug, Enzyme Induction, Enzyme Inhibitors toxicity, Female, Flutamide toxicity, Gas Chromatography-Mass Spectrometry, Herbicides toxicity, Liver drug effects, Liver enzymology, Male, Phenobarbital toxicity, Propylthiouracil toxicity, Rats, Rats, Wistar, Reproducibility of Results, Sex Factors, Tandem Mass Spectrometry, Time Factors, Toxicology standards, Tyrosine blood, Biomarkers blood, Metabolic Networks and Pathways drug effects, Systems Biology, Toxicology methods
- Abstract
Will metabolomics have a greater chance of success in toxicology and biomarker assessment than genomics and proteomics? Metabolomics has the advantage that (1) it analyses the last step in a series of changes following a toxic insult, (2) many of the metabolites have a known function and (3) changes are detectable in blood. If the analysis of a great number of individual organs can be replaced by one matrix then this will provide significant advantages (less invasive method, no need to kill animals, time course analysis possible). We have chosen to perform the analysis of blood metabolites in such a way as to minimize the risk of artifacts and to have a high number of known metabolites. We have also reduced the amount of variation in the biological system as well as during analysis. In a series of proof of concept studies it could be demonstrated that (1) the metabolome of control animals was stable of a period of nearly 1 year, with a remarkable differentiation between males and females, (2) a dose response relationship in metabolome changes was induced by phenobarbital and that (3) different modes of action could be distinguished by blood metabolome analysis. To investigate the potential of metabolomics to find biomarkers or specific patterns of change we have analyzed the blood metabolome of rats treated with HPPD inhibitors, a novel class of herbicides. The results demonstrated that a single metabolite, tyrosine, can be used as a biomarker. In addition to tyrosine we also found a specific pattern of change that involved nine metabolites. Though the extent of change was less than for tyrosine the consistent change of these metabolites is diagnostic for this (toxicological) mode of action.
- Published
- 2007
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28. Sedative and anticonvulsant effects of hydroalcoholic extract of Equisetum arvense.
- Author
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Dos Santos JG Jr, Blanco MM, Do Monte FH, Russi M, Lanziotti VM, Leal LK, and Cunha GM
- Subjects
- Animals, Anticonvulsants administration & dosage, Anticonvulsants therapeutic use, Dose-Response Relationship, Drug, Female, Hypnotics and Sedatives administration & dosage, Hypnotics and Sedatives therapeutic use, Male, Pentylenetetrazole, Plant Extracts administration & dosage, Plant Extracts therapeutic use, Plant Stems, Rats, Rats, Wistar, Seizures chemically induced, Anticonvulsants pharmacology, Equisetum, Hypnotics and Sedatives pharmacology, Maze Learning drug effects, Phytotherapy, Plant Extracts pharmacology, Seizures prevention & control, Sleep drug effects
- Abstract
The hydroalcoholic extract of Equisetum arvense (HAE) tested at the doses of 200 and 400 mg/kg showed a significant activity on the open-field, enhanced the number of falls in the rota-rod reducing the time of permanence in the bar and increased the sleeping time (46% and 74%) in the barbiturate-induced sleeping time. In the pentylenetetrazole-seizure, it increased the first convulsion latency, diminished the severity of convulsions, reduced the percentage of animals which developed convulsion (50% and 25%) and protected animals from death. On the contrary, in the elevated plus maze, the doses 50, 100 and 150 mg/kg did not affect the evaluated parameters. Thus, HAE presented anticonvulsant and sedative effects. Phytochemical analysis detected the presence of tannins, saponins, sterols and flavonoids.
- Published
- 2005
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29. Urokinase plasminogen activator amino-terminal peptides inhibit development of the rat ventral prostate.
- Author
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Elfman F, Bok R, Conn M, Shuman M, and Cunha G
- Subjects
- Animals, Animals, Newborn, Apoptosis physiology, Down-Regulation, Humans, Male, Mice, Organ Specificity, Peptides metabolism, Prostate ultrastructure, Protein Structure, Tertiary physiology, RNA, Messenger metabolism, Rats, Rats, Inbred F344, Receptors, Cell Surface antagonists & inhibitors, Receptors, Cell Surface genetics, Receptors, Urokinase Plasminogen Activator, Prostate growth & development, Receptors, Cell Surface physiology, Urokinase-Type Plasminogen Activator physiology
- Abstract
The plasma membrane urokinase plasminogen activator receptor (uPAR) localizes and enhances activation of pro-uPA. Active uPA, in turn, promotes increased degradation of the extracellular matrix (ECM) by activation of plasminogen. uPAR binds to ECM molecules and integrins, which can affect cellular adhesion, signal transduction, and gene regulation. The current study examines the expression and function of uPAR in developing rat ventral prostates (VPs). We report that newborn VPs express uPAR mRNA and protein. In addition, the function of uPAR-bound uPA during in vitro prostatic development was studied by adding recombinant peptide competitive inhibitors of uPA-uPAR binding. Newborn VP explants were cultured in serum-free media for one week with 10(-8) M testosterone plus chimeric peptides containing a human immunoglobulin G Fc domain and either human uPA amino acids 1-138 (hu-uPA 1-138) as a control or mouse uPA amino acids 1-138 (mo-uPA 1-138) or 1-48 (mo-uPA 1-48). Hu-uPA 1-138-treated VPs underwent normal ductal branching morphogenesis and tissue differentiation. In contrast, VPs treated with mo-uPA 1-138 or mo-uPA 1-48 displayed a dose-dependent perturbation of ductal branching. Differentiation of both epithelial and mesenchymal tissues was also impaired. Mo-uPA 1-48-treated VPs contained significantly more apoptotic cells. These observations suggest that disruption of uPA binding to uPAR results in a retardation of the development of newborn VPs.
- Published
- 2001
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30. Epithelial-stromal tissue interaction in paramesonephric (Müllerian) epithelial differentiation.
- Author
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Kurita T, Cooke PS, and Cunha GR
- Subjects
- Animals, CCAAT-Enhancer-Binding Protein-beta genetics, Estradiol pharmacology, Estrogen Receptor alpha, Female, Gene Expression Regulation, Developmental, Keratins genetics, Male, Mice, Mice, Knockout, Receptors, Estrogen genetics, Species Specificity, Uterus cytology, Uterus embryology, Uterus metabolism, Vagina cytology, Vagina embryology, Vagina metabolism, Cell Differentiation drug effects, Epithelial Cells cytology, Mullerian Ducts cytology, Stromal Cells cytology
- Abstract
During organogenesis, the middle to caudal portion of Müllerian epithelium differentiates into uterine and vaginal epithelia in females. Functional differentiation of uterine and vaginal epithelia occurs in adulthood, and is regulated by 17beta-estradiol (E(2)) and progesterone. In this report, the roles of mesenchyme/stroma in differentiation of uterine and vaginal epithelia were studied in tissue recombination experiments. At birth, Müllerian epithelium was negative for uterine and vaginal epithelial markers. Tissue recombinant experiments showed that uterine and vaginal gene expression patterns were induced in neonatal Müllerian epithelium by the respective mesenchymes. Differentiated adult uterine and vaginal epithelia did not change their original gene expression in response to heterotypic mesenchymal induction. In the adult vagina, E(2) induced expression of involucrin, a CCAAT/enhancer-binding protein beta and cytokeratin 1 via estrogen receptor alpha (ERalpha). Tissue recombination experiments with wild-type and ERalpha knockout mice demonstrated that epithelial gene expression is regulated by E(2) via epithelial-stromal tissue interactions. Uterine/vaginal heterotypic tissue recombinations demonstrated that functional differentiation of uterine and vaginal epithelia required organ-specific stromal factors. In contrast, stromal signals regulating epithelial proliferation appeared to be nonspecific in the uterus and vagina.
- Published
- 2001
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31. Cell differentiation lineage in the prostate.
- Author
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Wang Y, Hayward S, Cao M, Thayer K, and Cunha G
- Subjects
- Animals, Biomarkers, Glutathione S-Transferase pi, Glutathione Transferase analysis, Humans, Isoenzymes analysis, Keratins analysis, Male, Mice, Mice, Inbred BALB C, Rats, Rats, Sprague-Dawley, Stem Cells physiology, Cell Differentiation, Cell Lineage, Prostate embryology
- Abstract
Prostatic epithelium consists mainly of luminal and basal cells, which are presumed to differentiate from common progenitor/stem cells. We hypothesize that progenitor/stem cells are highly concentrated in the embryonic urogenital sinus epithelium from which prostatic epithelial buds develop. We further hypothesize that these epithelial progenitor/stem cells are also present within the basal compartment of adult prostatic epithelium and that the spectrum of differentiation markers of embryonic and adult progenitor/stem cells will be similar. The present study demonstrates that the majority of cells in embryonic urogenital sinus epithelium and developing prostatic epithelium (rat, mouse, and human) co-expressed luminal cytokeratins 8 and 18 (CK8, CK18), the basal cell cytokeratins (CK14, CK5), p63, and the so-called transitional or intermediate cell markers, cytokeratin 19 (CK19) and glutathione-S-transferase-pi (GSTpi). The majority of luminal cells in adult rodent and human prostates only expressed luminal markers (CK8, CK18), while the basal epithelial cell compartment contained several distinct subpopulations. In the adult prostate, the predominant basal epithelial subpopulation expressed the classical basal cell markers (CK5, CK14, p63) as well as CK19 and GSTpi. However, a small fraction of adult prostatic basal epithelial cells co-expressed the full spectrum of basal and luminal epithelial cell markers (CK5, CK14, CK8, CK18, CK19, p63, GSTpi). This adult prostatic basal epithelial cell subpopulation, thus, exhibited a cell differentiation marker profile similar to that expressed in embryonic urogenital sinus epithelium. These rare adult prostatic basal epithelial cells are proposed to be the progenitor/stem cell population. Thus, we propose that at all stages (embryonic to adult) prostatic epithelial progenitor/stem cells maintain a differentiation marker profile similar to that of the original embryonic progenitor of the prostate, namely urogenital sinus epithelium. Adult progenitor/stem cells co-express both luminal cell, basal cell, and intermediate cell markers. These progenitor/stem cells differentiate into mature luminal cells by maintaining CK8 and CK18, and losing all other makers. Progenitor/stem cells also give rise to mature basal cells by maintaining CK5, CK14, p63, CK19, and GSTpi and losing K8 and K18. Thus, adult prostate basal and luminal cells are proposed to be derived from a common pleuripotent progenitor/stem cell in the basal compartment that maintains its embryonic profile of differentiation markers from embryonic to adult stages.
- Published
- 2001
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32. Regulation of prostate branching morphogenesis by activin A and follistatin.
- Author
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Cancilla B, Jarred RA, Wang H, Mellor SL, Cunha GR, and Risbridger GP
- Subjects
- Activin Receptors, Activins, Animals, Follistatin, Glycoproteins analysis, Immunohistochemistry, Inhibins analysis, Inhibins genetics, Male, Morphogenesis drug effects, Organ Culture Techniques, Peptides analysis, RNA, Messenger analysis, Rats, Rats, Sprague-Dawley, Receptors, Growth Factor analysis, Glycoproteins pharmacology, Inhibin-beta Subunits, Inhibins pharmacology, Prostate embryology
- Abstract
Ventral prostate development occurs by branching morphogenesis and is an androgen-dependent process modulated by growth factors. Many growth factors have been implicated in branching morphogenesis including activins (dimers of beta(A) and beta(B) subunits); activin A inhibited branching of lung and kidney in vitro. Our aim was to examine the role of activins on prostatic development in vitro and their localization in vivo. Organ culture of day 0 rat ventral prostates for 6 days with activin A (+/- testosterone) inhibited prostatic branching and growth without increasing apoptosis. The activin-binding protein follistatin increased branching in vitro in the absence (but not presence) of testosterone, suggesting endogenous activins may reduce prostatic branching morphogenesis. In vivo, inhibin alpha subunit was not expressed until puberty, therefore inhibins (dimers of alpha and beta subunits) are not involved in prostatic development. Activin beta(A) was immunolocalized to developing prostatic epithelium and mesenchymal aggregates at ductal tips. Activin beta(B) immunoreactivity was weak during development, but was upregulated in prostatic epithelium during puberty. Activin receptors were expressed throughout the prostatic epithelium. Follistatin mRNA and protein were expressed throughout the prostatic epithelium. The in vitro evidence that activin and follistatin have opposing effects on ductal branching suggests a role for activin as a negative regulator of prostatic ductal branching morphogenesis., (Copyright 2001 Academic Press.)
- Published
- 2001
- Full Text
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33. The BMP family member Gdf7 is required for seminal vesicle growth, branching morphogenesis, and cytodifferentiation.
- Author
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Settle S, Marker P, Gurley K, Sinha A, Thacker A, Wang Y, Higgins K, Cunha G, and Kingsley DM
- Subjects
- Animals, Antigens, Differentiation, Bone Morphogenetic Protein Receptors, Type I, Cell Differentiation, Embryonic Induction, Epithelium, Growth Differentiation Factors, Growth Substances genetics, In Vitro Techniques, Infertility, Male genetics, Male, Mesoderm transplantation, Mice, Mice, Mutant Strains, Morphogenesis, Protein Serine-Threonine Kinases isolation & purification, Rats, Receptors, Growth Factor isolation & purification, Seminal Vesicles pathology, Bone Morphogenetic Proteins, Growth Substances metabolism, Seminal Vesicles embryology, Seminal Vesicles growth & development
- Abstract
Epithelial-mesenchymal interactions play an important role in the development of many different organs and tissues. The secretory glands of the male reproductive system, including the prostate and seminal vesicles, are derived from epithelial precursors. Signals from the underlying mesenchyme are required for normal growth, branching, and differentiation of the seminal vesicle epithelium. Here, we show that a member of the BMP family, Gdf7, is required for normal seminal vesicle development. Expression and tissue recombination experiments suggest that Gdf7 is a mesenchymal signal that acts in a paracrine fashion to control the differentiation of the seminal vesicle epithelium., (Copyright 2001 Academic Press.)
- Published
- 2001
- Full Text
- View/download PDF
34. fucosyltransferase1 and H-type complex carbohydrates modulate epithelial cell proliferation during prostatic branching morphogenesis.
- Author
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Marker PC, Stephan JP, Lee J, Bald L, Mather JP, and Cunha GR
- Subjects
- Androgens, Animals, Animals, Newborn, Antibodies, Monoclonal, Base Sequence, Cell Division, Cell Line, Fucosyltransferases genetics, Fucosyltransferases immunology, Humans, Lectins, Male, Molecular Sequence Data, Morphogenesis, Organ Culture Techniques, Paracrine Communication, Prostate cytology, RNA, Messenger isolation & purification, Rats, Rats, Sprague-Dawley, Galactoside 2-alpha-L-fucosyltransferase, Antigens, Differentiation metabolism, Epithelial Cells cytology, Fucosyltransferases metabolism, Plant Lectins, Prostate growth & development
- Abstract
The prostate undergoes branching morphogenesis dependent on paracrine interactions between the prostatic epithelium and the urogenital mesenchyme. To identify cell-surface molecules that function in this process, monoclonal antibodies raised against epithelial cell-surface antigens were screened for antigen expression in the developing prostate and for their ability to alter development of prostates grown in serum-free organ culture. One antibody defined a unique expression pattern in the developing prostate and inhibited growth and ductal branching of cultured prostates by inhibiting epithelial cell proliferation. Expression cloning showed that this antibody binds fucosyltransferase1, an alpha-(1,2)-fucosyltransferase that synthesizes H-type structures on the complex carbohydrate modifications of some proteins and lipids. The lectin UEA I that binds H-type 2 carbohydrates also inhibited development of cultured prostates. These data demonstrate a previously unrecognized role for fucosyltransferase1 and H-type carbohydrates in controlling the spatial distribution of epithelial cell proliferation during prostatic branching morphogenesis. We also show that fucosyltransferase1 is expressed by epithelial cells derived from benign prostatic hyperplasia or prostate cancer; thus, fucosyltransferase1 may also contribute to pathological prostatic growth. These data further suggest that rare individuals who lack fucosyltransferase1 (Bombay phenotype) should be investigated for altered reproductive function and/or altered susceptibility to benign prostatic hyperplasia and prostate cancer., (Copyright 2001 Academic Press.)
- Published
- 2001
- Full Text
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35. Evidence that epithelial and mesenchymal estrogen receptor-alpha mediates effects of estrogen on prostatic epithelium.
- Author
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Risbridger G, Wang H, Young P, Kurita T, Wang YZ, Lubahn D, Gustafsson JA, and Cunha G
- Subjects
- Animals, Animals, Newborn, Epithelial Cells cytology, Epithelial Cells drug effects, Estrogen Receptor alpha, Estrogen Receptor beta, Keratins genetics, Male, Mesoderm cytology, Metaplasia, Mice, Mice, Knockout, Mice, Transgenic, Prostate cytology, Prostate pathology, Receptors, Estrogen deficiency, Receptors, Estrogen genetics, Receptors, Progesterone genetics, Stromal Cells cytology, Stromal Cells pathology, Stromal Cells physiology, Up-Regulation, Diethylstilbestrol pharmacology, Epithelial Cells physiology, Mesoderm physiology, Prostate physiology, Receptors, Estrogen physiology
- Abstract
In combination with androgens, estrogens can induce aberrant growth and malignancy of the prostate gland. Estrogen action is mediated through two receptor subtypes: estrogen receptors alpha (ERalpha) and beta (ERbeta). Wild-type (wt) and transgenic mice lacking a functional ERalpha (alphaERKO) or ERbeta (betaERKO) were treated with the synthetic estrogen diethylstilbestrol (DES). DES induced prostatic squamous metaplasia (SQM) in wt and betaERKO but not in alphaERKO mice, indicating an essential role for ERalpha, but not ERbeta, in the induction of SQM of prostatic epithelium. In order to determine the respective roles of epithelial and stromal ERalpha in this response, the following tissue recombinants were constructed with prostatic epithelia (E) and stroma (S) from wt and ERKO mice: wt-S+wt-E, alphaERKO-S+alphaERKO-E, wt-S+alphaERKO-E, and alphaERKO-S+wt-E. A metaplastic response to DES was observed in wt-S+wt-E tissue recombinants. This response to DES involved multilayering of basal epithelial cells, expression of cytokeratin 10, and up-regulation of the progesterone receptor. Tissue recombinants containing alphaERKO-E and/or -S (alphaERKO-S+alphaERKO-E, wt-S+alphaERKO-E, and alphaERKO-S+wt-E) failed to respond to DES. Therefore, full and uniform epithelial SQM requires ERalpha in the epithelium and stroma. These results provide a novel insight into the cell-cell interactions mediating estrogen action in the prostate via ERalpha., (Copyright 2001 Academic Press.)
- Published
- 2001
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36. Plasticity of the urothelial phenotype: effects of gastro-intestinal mesenchyme/stroma and implications for urinary tract reconstruction.
- Author
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Li Y, Liu W, Hayward SW, Cunha GR, and Baskin LS
- Subjects
- Animals, Animals, Newborn, Cell Differentiation, Cell Transplantation, Fetal Tissue Transplantation physiology, Immunohistochemistry, Keratins analysis, Membrane Glycoproteins analysis, Mice, Mice, Inbred BALB C, Mucins analysis, Phenotype, Rats, Rats, Sprague-Dawley, Rectum embryology, Rectum growth & development, Stromal Cells cytology, Stromal Cells physiology, Urinary Tract surgery, Mesoderm cytology, Mesoderm physiology, Urothelium cytology, Urothelium physiology
- Abstract
The present study tests the hypothesis that heterotypic stromal-epithelial interactions cause phenotypic changes in urothelium. The rational for the experimental design is to simulate heterotypic stromal-epithelial interactions that are created at the anastomotic site of intestinal-bladder augmentations and internal urinary diversions where the urothelium is in direct contact with the gastro-intestinal tract tissues. Tissue recombination experiments were performed by combining 14-day embryonic rat and mouse rectal mesenchyme with urothelium from embryonic, newborn, and adult mice or rats. All tissue recombinants were grown beneath the renal capsule of athymic mouse hosts for 6-16 weeks. Analyses were performed to detect expression of uroplakins, cytokeratin 7, 14, 19 and mucin secreting epithelial cells via Periodic Acid-Schiff (PAS). The phenotype of both mouse and rat urothelium was changed to a glandular morphology under the influence of rectal mesenchyme. Immunohistochemical staining revealed a loss of the urothelial specific uroplakins and cytokeratins 7, 14, and 19 (characteristic of urothelium). Histologic analysis revealed the presence of mucin secreting glandular structures which stained positive for PAS. The urothelial transdifferentiation into glandular epithelium was not a function of epithelial age and occurred in the embryonic, newborn and adult urothelium. Likewise, rectal mesenchyme from embryonic, neonatal, and adult animals was able to induce glandular differentiation in bladder epithelium. Urothelium exhibits the plasticity to change into an intestinal like epithelium as a result of mesenchymal/stromal stimulation from the gastro-intestinal tract. This experimental result is germane to heterotypic stromal-epithelial interactions that are created in patients with urinary tract reconstructions (intestinal augmentations, de-mucosalized urothelial lined bladder patches, and internal urinary diversion such as ureterosigmoidostomies). We propose that heterotypic stromal-epithelial interactions may play a role in determining histodifferentiation of urothelial cells at the anastomotic site between bowel and bladder tissue in patients with gastro-intestinal urothelial reconstructions.
- Published
- 2000
- Full Text
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37. Pentoxifylline improves learning and memory in glutamate-lesioned rats.
- Author
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Cunha GM, Bezerra PJ, Saldanha MD, Cavalcante MC, De Brun VM, and Viana GS
- Subjects
- Animals, Avoidance Learning drug effects, Cyclic AMP metabolism, Hippocampus drug effects, Male, Maze Learning drug effects, Rats, Rats, Wistar, Receptors, N-Methyl-D-Aspartate drug effects, Glutamic Acid toxicity, Learning drug effects, Memory drug effects, Pentoxifylline pharmacology, Phosphodiesterase Inhibitors pharmacology, Receptors, Glutamate drug effects
- Abstract
The present work shows the effects of pentoxifylline (ptx), on learning and memory in rats with hippocampal lesions induced by glutamate (glu). Immediately after stereotaxic procedures and in the absence or presence of glu lesions, animals were treated with ptx (50, 100, or 200 mg/kg, IP) for 6 days. Twenty-four hours after the last injection, behavior and memory tests were performed, animals were sacrificed, and hippocampi dissected for cAMP determination or histopathological studies. Results from the T-maze task showed a less learning ability in the glu-lesioned group compared to other ones. Ptx alone or associated with glu significantly improved memory acquisition, but not memory consolidation compared to glu-lesioned rats. Except for the increased locomotor activity observed in the ptx100+glu-treated group compared to saline, no other difference was detected in the open-field test. A significant impairment in avoidance performance was observed in glu-lesioned group as compared to saline or to other groups in the short as well as in the late phase of memory. All groups showed an improved water-maze performance over time with similar performances on the final day of acquisition. The impairment in memory retention observed in glu-lesioned rats was reversed by the pretreatment with ptx200. Glu induced hippocampal lesion and reduced cAMP levels. Both effects were blocked by ptx, suggesting that its action may be the result of increased cAMP levels and/or inhibition of adenosine A1 receptors.
- Published
- 2000
- Full Text
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38. Mesenchymal reprogramming of adult human epithelial differentiation.
- Author
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Aboseif S, El-Sakka A, Young P, and Cunha G
- Subjects
- Adult, Animals, Animals, Newborn, Antigens, Differentiation metabolism, Blotting, Western, Cell Transplantation, Cells, Cultured, Coculture Techniques, Epithelial Cells metabolism, Humans, Immunohistochemistry, Kidney cytology, Male, Mesoderm transplantation, Mice, Mice, Nude, Rats, Rats, Sprague-Dawley, Seminal Vesicles cytology, Seminal Vesicles transplantation, Stromal Cells cytology, Stromal Cells metabolism, Urinary Bladder transplantation, Cell Differentiation, Epithelial Cells cytology, Mesoderm cytology, Urinary Bladder cytology
- Abstract
The objective of this study was to determine whether neonatal rat seminal vesicle mesenchyme (rSVM) can reprogram epithelial differentiation in a fully differentiated adult human bladder epithelium. For this purpose neonatal rSVM was isolated from newborn (0-day) Sprague-Dawley rats, and normal adult human bladder epithelium (hBLE) was isolated from radical cystoprostatectomy specimens to prepare rSVM+hBLE tissue recombinants in vitro. After overnight culture the tissue recombinants were grafted beneath the renal capsule of male athymic rodent hosts and allowed to grow in vivo for 6 months. As controls, rSVM and hBLE were grafted separately and allowed to grow for the same period. Tissue recombinants and control tissue grafts were harvested, and secretions were collected for biochemical studies. Tissues were fixed both for histologic as well as immunohistochemical staining. Neonatal rSVM induced normal adult human bladder urothelium to form glandular structures resembling prostate. The induced prostatic acini were filled with secretions that expressed human prostate-specific secretory proteins. These findings demonstrate that adult human urothelial cells retain a responsiveness to neonatal prostatic mesenchymal inductors. Change in urothelial histodifferentiation was associated with change in functional activity. The ability of the neonatal rat mesenchymal tissues to induce morphologic as well as biochemical changes in normal adult human urothelium provides a basis for human tissue engineering and organ reconstruction.
- Published
- 1999
- Full Text
- View/download PDF
39. Ontogeny of the male urethra: theory of endodermal differentiation.
- Author
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Kurzrock EA, Baskin LS, and Cunha GR
- Subjects
- Animals, Cell Differentiation physiology, Embryonic Induction physiology, Epithelium transplantation, Humans, Immunohistochemistry methods, Keratins analysis, Keratins metabolism, Kidney surgery, Male, Mesoderm cytology, Mesoderm transplantation, Mice, Mice, Nude, Rats, Tissue Transplantation, Transplantation, Heterologous, Urinary Bladder cytology, Urothelium cytology, Urothelium embryology, Endoderm cytology, Urethra cytology, Urethra embryology
- Abstract
The most widely accepted mechanism of male urethral development proposes that the urethral plate is elevated by urethral folds which fuse ventrally in a proximal-to-distal sequence. Unlike its proximal counterpart, the urethra which forms within the glans is lined by a stratified squamous epithelium and has a more controversial development. One theory supports the idea that fusion of the urethral folds extends all the way to the tip of the glans. Another theory suggests that a solid ectodermal in-growth of epidermis canalizes the glandar urethra. We hypothesized that the use of immunohistochemical staining and tissue recombinant grafting would delineate the epithelia involved and lend clues to their origin. Thirty-six human fetal phallic specimens of gestational ages 5-22 weeks were sectioned and stained immunohistochemically with antibodies raised against different cytokeratins. Evaluation of the sections showed that the urethral plate, an extension of the urogenital sinus, extended to the tip of the phallus and maintained patency and continuity throughout the process of urethral development. The entire urethra, including the glans portion, was formed by dorsal extension and disintegration of the urethral plate combined with ventral growth and fusion of the urethral folds. Sections of the distal glandar urethra showed no evidence of a solid ectodermal ingrowth. Rather, immunostaining results at different ages suggested differentiation of the endodermal urethral plate into a stratified squamous epithelium. To determine whether urothelium could be induced to express a stratified squamous phenotype, mouse fetal bladder epithelium was combined with rat fetal genital tubercle mesenchyme and grown under the renal capsule of athymic mice. The bladder epithelium differentiated into a stratified squamous epithelium. Thus, proper mesenchymal signaling may induce differentiation of urothelium into a stratified squamous phenotype, such as during development of the urethra of the glans penis.
- Published
- 1999
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- View/download PDF
40. Stromal cells are critical targets in the regulation of mammary ductal morphogenesis by parathyroid hormone-related protein.
- Author
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Dunbar ME, Young P, Zhang JP, McCaughern-Carucci J, Lanske B, Orloff JJ, Karaplis A, Cunha G, and Wysolmerski JJ
- Subjects
- Animals, Binding, Competitive, Cyclic AMP pharmacology, Embryonic and Fetal Development genetics, Epithelium physiology, Female, In Situ Hybridization, Mesoderm physiology, Mice, Mice, Inbred Strains, Parathyroid Hormone-Related Protein, RNA, Messenger genetics, Receptors, Parathyroid Hormone genetics, Gene Expression Regulation, Developmental genetics, Mammary Glands, Animal growth & development, Morphogenesis physiology, Proteins physiology, Stromal Cells physiology
- Abstract
Parathyroid hormone-related protein (PTHrP) was originally identified as the tumor product responsible for humoral hypercalcemia of malignancy. It is now known that PTHrP is produced by many normal tissues in which it appears to play a role as a developmental regulatory molecule. PTHrP is a normal product of mammary epithelial cells, and recent experiments in our laboratory have demonstrated that overexpression or underexpression of PTHrP in the murine mammary gland leads to severe disruptions in its development. The nature of these phenotypes suggests that PTHrP acts to modulate branching growth during mammary development by regulating mammary stromal cell function. We now demonstrate that throughout mammary development, during periods of active ductal-branching morphogenesis, PTHrP is produced by epithelial cells, whereas the PTH/PTHrP receptor is expressed on stromal cells. In addition, we show that mammary stromal cells in culture contain specific binding sites for amino terminal PTHrP and respond with an increase in intracellular cAMP. Finally, we demonstrate that the mammary mesenchyme must express the PTH/PTHrP receptor in order to support mammary epithelial cell morphogenesis. These results demonstrate that PTHrP and the PTH/PTHrP receptor represent an epithelial/mesenchymal signaling circuit that is necessary for mammary morphogenesis and that stromal cells are a critical target for PTHrP's action in the mammary gland., (Copyright 1998 Academic Press.)
- Published
- 1998
- Full Text
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41. Interactions between adult human prostatic epithelium and rat urogenital sinus mesenchyme in a tissue recombination model.
- Author
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Hayward SW, Haughney PC, Rosen MA, Greulich KM, Weier HU, Dahiya R, and Cunha GR
- Subjects
- Adult, Animals, Collagenases metabolism, Epithelium anatomy & histology, Epithelium physiology, Female, Humans, Hyaluronoglucosaminidase metabolism, In Situ Hybridization, Fluorescence, Male, Mice, Mice, Nude, Polymerase Chain Reaction, Pregnancy, Prostate physiology, Rats, Signal Transduction physiology, Species Specificity, Subrenal Capsule Assay, Transcription, Genetic, Urogenital System physiology, Mesoderm physiology, Prostate anatomy & histology, Urogenital System anatomy & histology
- Abstract
Tissue recombinants composed of adult human prostatic epithelium (hPrE) and rat urogenital sinus mesenchyme (rUGM) were grafted beneath the renal capsule of athymic rodent hosts. The pseudostratified human epithelium initially became multilayered, solid epithelial cords emerged, grew into the surrounding mesenchyme and canalized to regenerate a pseudostratified epithelium. Basal cells expressed cytokeratins 5 and 14, while luminal cells expressed cytokeratins 8 and 18, prostate specific antigen and prostatic acid phosphatase. The rat mesenchymal component differentiated into thick sheets of smooth muscle, characteristic of the human but not the rat prostate. These findings indicate that epithelial-mesenchymal interactions were reciprocal. Rat UGM induced adult hPrE to form new ductal-acinar tissue, involving epithelial proliferation, ductal branching morphogenesis and functional cytodifferentiation. Concurrently the epithelium dictated smooth muscle differentiation and patterning. Species-specific reverse transcriptase polymerase chain reaction SC (RT-PCR) analysis of the tissue recombinants was performed to separately examine the expression of epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha), epidermal growth factor receptor (EGFR), TGF-beta 1, and TGF-beta 3 in the epithelium, stroma and host components of the graft. All of these genes, except TGF-beta 1, were expressed in all three tissues. Human TGF-beta 1 was not detected, indicating that this gene was not expressed in human prostatic epithelium but was present in stroma.
- Published
- 1998
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42. Enlargement of the ampullary gland and seminal vesicle, but not the prostate in int-2/Fgf-3 transgenic mice.
- Author
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Donjacour AA, Thomson AA, and Cunha GR
- Subjects
- Animals, Disease Models, Animal, Evaluation Studies as Topic, Female, Hypertrophy, Male, Mice, Mice, Transgenic, Organ Size physiology, Phenotype, Testis pathology, Urogenital System pathology, Gene Expression Regulation, Developmental physiology, Gene Expression Regulation, Neoplastic physiology, Prostatic Hyperplasia pathology, Seminal Vesicles pathology, Vas Deferens pathology
- Abstract
Expression of the int2/Fgf-3 gene occurs during normal embryonic development and is associated with mammary cancer in mice. Overexpression of this gene under the control of the mouse mammary tumor virus long terminal repeat (MMTV-LTR) in males was reported to result in prostatic enlargement. In this report male Fgf-3-overexpressing mice were shown to have enlarged ampullary glands, seminal vesicles, and ductus deferens; there was extensive epithelial hyperplasia in the ampullary glands and seminal vesicles. The prostates of these animals were of normal size and histology. The transgene was expressed in all of the enlarged organs, which are derived exclusively from the Wolffian duct. Male secondary sex organs derived from the urogenital sinus, e.g., the ventral prostate, coagulating gland, and bulbourethral glands, were normal and did not express the MMTV-LTR-driven Fgf-3 transgene. A dorsolateral prostate was also morphologically normal but did express the transgene. This study underscores the importance of careful organ identification in transgenic models in which gross organ enlargement or distortion occurs. It also highlights the heterogeneity of the response to Fgf-3 among the secondary sex organs and even within the prostate itself.
- Published
- 1998
- Full Text
- View/download PDF
43. Estrogen-induced epithelial proliferation and cornification are uncoupled in sinus vaginal epithelium associated with uterine stroma.
- Author
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Boutin EL and Cunha GR
- Subjects
- Animals, Cell Division drug effects, Culture Techniques methods, Diethylstilbestrol pharmacology, Epithelial Cells drug effects, Epithelial Cells metabolism, Female, Mesoderm cytology, Mesoderm drug effects, Mice, Mice, Inbred BALB C, Mice, Nude, Rats, Rats, Sprague-Dawley, Receptors, Estrogen drug effects, Receptors, Estrogen immunology, Receptors, Estrogen metabolism, Stromal Cells physiology, Time Factors, Uterus drug effects, Vagina drug effects, Vagina metabolism, Cell Differentiation drug effects, Estradiol pharmacology, Uterus cytology, Vagina cytology
- Abstract
Epithelium from the urogenital sinus-derived portion of the newborn mouse vagina when grown in association with uterine mesenchyme forms a "vaginal" stratified squamous non-cornified epithelium. However, the epithelium of these tissue recombinants composed of sinus vaginal epithelium plus uterine mesenchyme does not undergo the fluctuations in cytodifferentiation normally seen in vaginal epithelium during the estrous cycle (e.g., cornification and mucification). In this report we show that sinus vaginal epithelium in association with uterine mesenchyme proliferated in response to estradiol but failed to cornify in response to diethylstilbestrol (DES), even though both the epithelium and the stroma had estrogen receptors. However, if sinus vaginal epithelium that had been grown in combination with uterine mesenchyme was re-isolated from the tissue recombinant and recombined with fresh vaginal mesenchyme, the epithelium cornified in response to DES. These results indicate that the proliferative and the cytodifferentiation response to estrogen could be uncoupled and that sinus vaginal epithelium required vaginal stroma to cornify in response to DES.
- Published
- 1997
- Full Text
- View/download PDF
44. Decreased prostate cancer cell migration by inhibition of the insulin-like growth factor II/Mannose-6-Phosphate receptor.
- Author
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Evans CP, Elfman F, Cunha G, and Shuman MA
- Abstract
The 270-kDa insulin-like growth factor II (IGF-II)/cation-independent mannose-6-phosphate receptor (MPR) is a multifunctional receptor protein. Endocytoses and intracellular transport of soluble enzymes bearing mannose6-phosphate (M-6-P) residues to lysosomes is mediated by the IGF-II/MPR. We examined human prostate cancer cells for IGF-II/MPR expression to determine whether this receptor mediates cell migration. PC3 human prostate cancer cells were studied for intracellular IGF-II/MPR by immunoblotting. PC3 cell surface IGF-II/MPR expression was assessed by flow cytometric analysis. Cell motility was quantitated by a scratch migration assay, and IGF-II/MPR blockade was achieved using M-6-P or affinity-purified rabbit anti-bovine cation-independent IGF-II/MPR immunoglobulin. IGF-II/MPR is expressed in the cytoplasm and on the surface of PC3 prostate cancer cells. The mean number of PC3 cells migrating per high powered field in medium containing polyclonal anti-IGF-II/MPR immunoglobulin or M-6-P decreased significantly (5 ± 4 cells and 34 ± 5 cells, respectively) compared with control medium containing mouse immunoglobulin G (70 ± 12 cells) or mannose-1-phosphate (67 ± 7 cells). This decreased PC3 cell migration following cell surface IGF-II/MPR blockade suggests that the IGF-II/MPR may play an important role in prostate cancer cell motility.
- Published
- 1997
- Full Text
- View/download PDF
45. An edgewise look at basal epithelial cells: three-dimensional views of the rat prostate, mammary gland and salivary gland.
- Author
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Hayward SW, Brody JR, and Cunha GR
- Subjects
- Actins analysis, Animals, Cell Size, Epithelial Cells, Female, Immunohistochemistry, Keratins analysis, Male, Pregnancy, Rats, Rats, Sprague-Dawley, Basal Metabolism, Image Processing, Computer-Assisted, Mammary Glands, Animal cytology, Prostate cytology, Salivary Glands cytology
- Abstract
Wholemount immunocytochemical staining was used to visualize basal and luminal epithelial-cell-specific cytokeratin and smooth muscle alpha-actin expression in the developing and adult rat prostate, in the pregnant rat mammary gland and adult rat salivary gland. The stained samples were examined using an Edge R400 3D microscope. Images were collected in both single-image and stereo-pair formats. Prostatic basal epithelial cells were found to have a cell body covering an area of 52-62 microns 2. The mean footprint size of basal cells was not significantly different between prostatic lobes. Basal epithelial cells were most dense in the anterior and most sparse in the ventral prostatic lobes. Basal epithelial cells had a large body with many processes, which spread around the duct and projected between luminal cells towards the lumen. These processes closely approached their counterparts from adjacent basal cells. In the developing prostate the differentiation of the basal cells from undifferentiated epithelial cords was observed at the region of ductal widening associated with canalization. Following castration prostatic basal epithelial cells became more closely packed, though the size of individual cells was not significantly changed. There was a two-to four-fold increase in basal cell density by 7 days after surgery. Most prostatic luminal cells were found to have hexagonal bases though some were pentagonal in shape. Luminal cells had a mean basal area of 50 microns 2. In the prostate immunocytochemical staining against smooth muscle alpha-actin revealed discrete bands of muscle surrounding individual prostatic ducts. In the mammary and salivary glands the epithelium was organized into an alveolar arrangement. In the salivary gland a single basal epithelial cell covered the top of each alveolus with processes arranged down the side of the structure. In the mammary gland several basal cells were draped over each alveolus. The mammary and salivary gland basal cells expressed smooth muscle alpha-actin, indicating their myoepithelial phenotype. The organization of the mammary and salivary gland basal cells placed them in an ideal position to squeeze the alveolar structures.
- Published
- 1996
- Full Text
- View/download PDF
46. Urothelial transformation into functional glandular tissue in situ by instructive mesenchymal induction.
- Author
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Lipschutz JH, Young P, Taguchi O, and Cunha GR
- Subjects
- Animals, Cell Division, Epithelial Cells, Epithelium metabolism, Female, Male, Mesoderm metabolism, Mice, Mice, Inbred C57BL, Rats, Rats, Sprague-Dawley, Seminal Vesicles metabolism, Ureter metabolism, Mesoderm cytology, Receptors, Androgen metabolism, Seminal Vesicles cytology, Ureter cytology
- Abstract
It is generally believed that adult tissue is terminally differentiated. The ureter is derived from the metanephric diverticulum which, along with the derivatives of the metanephric mesoderm, forms the kidney. In our experiments, the left ureters of adult male athymic mouse hosts were severed below the kidney, and mesenchyme from neonatal rat seminal vesicles (SVM) was grafted to the cut end of the ureter, thus bringing adult mouse ureter epithelium (URE) in contact with neonatal rat SVM. After four to eight weeks, the in situ tissue recombinants were harvested, and the epithelial secretory proteins recovered. In 5 of 11 cases, an induction occurred, resulting in an in situ transformation of the non-glandular transitional epithelium of the adult mouse ureter into the simple columnar epithelium of the seminal vesicle (SV). Functional cytodifferentiation was examined in these neonatal rat SVM + adult mouse URE tissue recombinants using antibodies against SV-specific secretory (SVS) proteins of the mouse and rat. From the cut end of the ureter, the adult URE was induced to undergo SV morphogenesis, to express SV cytodifferentiation, and to produce the complete spectrum of major SVS proteins characteristic of the mouse. The induced seminal vesicle epithelium (SVE) also expressed androgen receptors (AR) which are not seen in urothelial tissue. Staining with Hoechst dye 33258, which can distinguish cells of mouse and rat origin, further demonstrated that the induced SVE was indeed of mouse origin and not a contaminant of the inducing rat SVM. in addition, neonatal mouse vaginal mesenchyme was grafted in situ beneath the bladder mucosa of adult male mice, and the host animals were killed after three months. The vaginal mesenchyme implanted into the bladders induced prostate-like acini, indicating that the above reprogramming of adult organs in situ is not an isolate occurrence. These results set a precedent for the "recreation" of new vital organs, such as the kidney, in situ by demonstrating that adult epithelial cells retain a developmental plasticity equivalent to their undifferentiated fetal counterparts and are capable of being reprogrammed in situ to express a completely new morphological, biochemical, and functional phenotype.
- Published
- 1996
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47. Upregulation of the 72-kDa type IV collagenase in epithelial and stromal cells during rat tracheal gland morphogenesis.
- Author
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Lim M, Elfman F, Dohrman A, Cunha G, and Basbaum C
- Subjects
- Animals, Base Sequence, Epithelium metabolism, Male, Matrix Metalloproteinase 9, Molecular Sequence Data, Molecular Weight, Morphogenesis, RNA, Messenger analysis, Rats, Rats, Inbred F344, Stromal Cells metabolism, Trachea enzymology, Up-Regulation, Collagenases genetics, Gene Expression Regulation, Developmental, Gene Expression Regulation, Enzymologic, Trachea growth & development
- Abstract
Submucosal glands secrete most of the mucus that lubricates the tracheal surface and protects it from irritants and infection. These glands develop postnatally in the rat, permitting convenient study of the mechanisms controlling this process. One such mechanism involves degradation of the supportive connective tissue matrix at the front of the growing glands. We recently showed that tracheal gland cell invasion of collagen gels in vitro is dependent on secretion of a 72-kDa type IV collagenase. In the present study, we show that the activity of this enzyme (also referred to as matrix metalloproteinase-2 or gelatinase A) is elevated at the time of gland development in vivo. That this increase is at least partly mediated at the level of steady-state mRNA was indicated by semiquantitative PCR analysis of gland-enriched, microdissected tissue samples. Immunohistochemistry revealed that the enzyme was present at the interface between the glands and extracellular matrix. In situ hybridization revealed that the cognate mRNA was present in epithelial cells of glands undergoing morphogenesis (particularly Postnatal Day 7) but not in those of adult glands or the surface epithelium. At all ages, stromal cells below the surface epithelium were labeled; labeling intensity was highest at the time and location of gland morphogenesis. These findings suggest that the 72-kDa type IV collagenase is developmentally regulated in gland and stromal cells at the level of steady-state mRNA and plays a role in the degradation of extracellular matrix during tracheobronchial gland morphogenesis.
- Published
- 1995
- Full Text
- View/download PDF
48. Terms and techniques: New approach to hot-start polymerase chain reaction using Taq DNA polymerase antibody.
- Author
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Dahiya R, Deng G, Chen K, Haughney PC, Cunha GR, and Narayan P
- Abstract
The purpose of this study was to optimize the conditions for polymerase chain reaction (PCR). The most common problem with conventional PCR is the presence of nonspecific products and primer-dimers formation, which could be due to several factors such as annealing temperature, primer concentration, and Taq DNA polymerase activity during setup of the PCR. Recently, a neutralizing monoclonal antibody (TaqStart) that blocks Taq DNA Polymerase activity has been developed. In the present study, we determined the effects of Taq DNA polymerase monoclonal antibody (TaqStart antibody) and other parameters, such as annealing temperature and oligonucleotide primer concentrations, on the specificity of PCR products. The results of these experiments suggest that TaqStart antibody inhibits nonspecific products and primer-dimers formation by blocking Taq DNA polymerase activity until the reaction components are heated in the thermal cycler (hot start). Other factors such as annealing temperature, oligonucleotide primer concentration, and magnesium concentration are equally important for specificity of PCR products.
- Published
- 1995
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- View/download PDF
49. Keratinocyte growth factor and acidic fibroblast growth factor are mitogens for primary cultures of mammary epithelium.
- Author
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Imagawa W, Cunha GR, Young P, and Nandi S
- Subjects
- Animals, Cell Division drug effects, Cells, Cultured, Culture Techniques methods, DNA biosynthesis, DNA Replication drug effects, Dose-Response Relationship, Drug, Epithelial Cells, Epithelium drug effects, Female, Fibroblast Growth Factor 10, Fibroblast Growth Factor 7, Kinetics, Mammary Glands, Animal drug effects, Mice, Mice, Inbred BALB C, Thymidine metabolism, Fibroblast Growth Factor 1 pharmacology, Fibroblast Growth Factors, Growth Substances pharmacology, Mammary Glands, Animal cytology, Mitogens pharmacology
- Abstract
Mammary epithelial cells derived from the entire mammary parenchyma or only end buds were isolated by collagenase digestion of mammary glands from virgin mice. Cells were cultured within collagen gels in serum-free medium containing insulin. Keratinocyte growth factor (KGF or FGF-7) and acidic fibroblast growth factor (aFGF or FGF-1) stimulated multifold proliferation when added alone to this medium. Growth occurred as three-dimensional colonies within the collagen gel matrix. KGF stimulated growth was unaffected by adding heparin. Conversely, multifold growth stimulation by acidic FGF required heparin. Since end buds are the actively proliferating cell population of ductal glands, organ cultures of these structures were prepared. KGF stimulated 3H-thymidine incorporation in these end buds in the absence and presence of epidermal growth factor. These data suggest that acidic FGF and KGF may represent in vivo stromal factors capable of regulating mammary gland development.
- Published
- 1994
- Full Text
- View/download PDF
50. An ultrastructural study of the developing urogenital tract in early human fetuses.
- Author
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Lawrence WD, Whitaker D, Sugimura H, Cunha GR, Dickersin GR, and Robboy SJ
- Subjects
- Cell Nucleolus ultrastructure, Cell Nucleus ultrastructure, Cytoplasm ultrastructure, Epithelium ultrastructure, Humans, Mesonephros ultrastructure, Microscopy, Electron, Mitosis, Urogenital System embryology, Urogenital System ultrastructure
- Abstract
Objectives: This study examines the gonoducts during the ambisexual stage of human fetal development to define their ultrastructural characteristics, including the origin and antomic relationship of the mesonephric and paramesonephric ducts during gonoductal development., Study Design: The reproductive tracts from five fetuses ranging in gestational age from 35 to 45 days were processed for ultrastructural examination. The developing mesonephric ducts, paramesonephric ducts, and their surrounding mesenchyme were studied with a Phillips 300 transmission electron microscope., Results: The mesonephric ducts and paramesonephric ducts have distinctive cytoplasmic and cell surface ultrastructural characteristics, as well as different morphologic patterns of epithelial-mesenchymal interaction. Cephalad portions of mesonephric ducts and paramesonephric ducts are separated by mesenchyme, but more caudal aspects move progressively closer until they are juxtaposed but separate., Conclusions: Early mesonephric ducts and paramesonephric ducts may be distinguished because of their distinctive ultrastructural features; epithelial-mesenchymal interaction may be important in their differentiation and maintenance; both gonoducts retain their morphologic identity throughout, supporting their separate origins.
- Published
- 1992
- Full Text
- View/download PDF
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