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Background: Flow cytometry protocols for counting fresh CD34+ cell samples are not ideal for cryopreserved products due to cryoprotectant cytotoxicity. For cryopreserved samples, often large volumes of hypotonic solutions, which can cause cell death, are used to remove the cryoprotectant with a post-thaw wash. We recently developed a novel multistep dilution method with subsequent flow cytometry analysis to allow for accurate and reproducible results. The previous method involved washing steps which invalidate the ability to enumerate cell recovery, and success had to be gauged solely on viability. The new method allows for assessment of total cell recovery and viable cell recovery., Study Design and Methods: Apheresis products were cryopreserved in 10% DMSO at a target WBC concentration of 300 × 10 6 /mL. Cryovials from these products were thawed at 37°C, and samples were diluted 1:2 by three additions of 1/3 sample volume using 1%-Human Albumin in Dextran 40 (10% Low Molecular Weight Dextran in 0.9% NaCl) separated by 5 min between each addition. A 1:10 dilution was performed to obtain the correct cell concentration for flow cytometric analysis resulting in a 1:20 dilution. End WBC concentrations were ~15 × 10 6 /mL with DMSO diluted to 0.5%., Results: Fifty-two samples were tested with this new method. Total and viable cell recoveries were calculated based on pre-cryopreservation data. Median total cell recoveries for CD34 and CD3 were >85%, while median viable cell recoveries were >75%., Discussion: Cryopreserved samples can be reliably prepared for flow cytometric testing using a step-wise dilution to preserve cell integrity and robust recoveries., (© 2024 AABB.)

Bioactives derived from marine sources are opening up new therapeutic opportunities for neurosurgery. These include tissue regeneration, neuroprotection, and infection control. These molecules derived from extreme marine conditions have generated new modes of action, for example, selective inhibition of voltage-gated ion channels and influencing pathways of neuroinflammation. Representative examples include chitosan for nerve regeneration, conotoxins for pain modulation, and fucoidan for neuroprotection. Despite therapeutic promise, major challenges remain: those of sustainable sourcing and regulatory validation. This letter points up the potential of marine bioactives to change neurosurgical care and challenges the brain communities to work together across disciplines in order to make that happen., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: Standard flow cytometry protocols for CD34+ cell enumeration designed for fresh samples are not appropriate for cryopreserved products. Special protocols have been developed to remove the cryoprotectant by quickly washing a freshly thawed sample. Exposing cells to a large volume of hypotonic solution and subsequent washing process was hypothesized to cause lab-induced cell death. Moreover, standard gating strategies must be altered to avoid reporting falsely high viabilities., Study Design and Methods: We developed a novel method whereby thawed samples were diluted step-wise to 1:2 by 3 additions of 1/3 sample volume using 1% Human Albumin in Dextran 40 (10% Low Molecular Weight Dextran in 0.9% NaCl) separated by 5 min between each addition. An additional 1:10 dilution was required to obtain a desired cell concentration for flow cytometry testing resulting in a 1:20 dilution., Results: Twenty samples were tested simultaneously in a method comparison; the new method demonstrated significant increases in mean cell viabilities for white blood cells, hematopoietic progenitor cells, and T cells as well as reduced standard deviations for each parameter., Discussion: Slow, step-wise dilutions of freshly thawed samples of cryopreserved apheresis products to 1:20 yielded higher and more precise viability measurements compared to quickly washing samples to remove DMSO., (© 2024 AABB.)

Objective: The objective of the experiment was to study yeast supplementation (yeast, Y) and dried kratom leaves (DKTL) on the digestibility, ruminal fermentation, blood metabolites and nitrogen balance in goats., Methods: Four of 7 to 8 months old male crossbred (50% Thai Native-Anglo Nubian) goats with average liveweight 20±0.13 kg were randomly assigned according to a 2×2 factorial arrangement in a 4×4 Latin square design to receive four diets ad libitum basis. The study investigated the effects of two levels of yeast (Y) supplementation (Y, 0 and 0.5g/kg dry matter [DM]) along with two levels of DKTL supplementation (DKTL, 0 and 4.44g/kg DM). The experimental groups were as follows: T1 = control group with 0Y+0DKTL, T2 = 0Y+4.44 DKTL, T3 = 0.5Y+0DKTL, and T4 = 0.5Y+4.44 DKTL., Results: The results showed that there were no interactions between Y levels and DKTL levels with respect to total DM intake, but there were significant effects (p<0.05) by levels of Y; goats receiving 0.05 g/kg DM Y had higher than goats fed 0.0 g/kg DM on average (kg/d). A percentage of body weight (% BW) and grams per kilogram of metallic weight (g/kg w0.75) had no influence on yeast levels and DKTL, but there was a difference (p<0.05) by yeast level Y at 0.5 g/kg DM, being higher compared to the non-supplemented group. Apparent digestibility coefficient of nutrition in the form of (DM, organic matter, crude protein, neutral detergent fiber, and acid detergent fiber) was an increased trend in the Y-level complementary group at 0.5 g/kg DM and DKTL at 4.44 g/kg DM, respectively. Protozoa populations decreased in the group receiving Y levels at 0.5 g/kg DM and DKTL levels at 4.44 g/kg DM when compared to group T1. The acetic acid concentration and methane gas generation decreased (p<0.05) in the group receiving Y levels of 0.5 g/kg DM and DKTL levels of 4.44 g/kg DM, while the amount of propionic acid increased (p<0.05)., Conclusion: Effects of feeding combinations of Y and DKTL supplementation on feed showed no interaction effect (Y×DKTL) on feed intake, rumen fermentation, bacterial and fungi population. The effect on protozoal populations was lower in the group that was supplemented with DKTL at 4.44 g/kg DM related to synthetic CH4 was reduced.

The overuse of exclusionary discipline practices, such as out-of-school suspensions (OSS), has consistently been documented over several decades. The resulting racial discipline disparities and the negative outcomes related to OSS have led policy makers and educators to consider other approaches to school discipline. One such approach, which has gained increasing use in the USA, is restorative practices (RP). However, despite its popularity, the experimental evidence base of the effects of RP and suspensions is extremely limited. To add to this knowledge base, we present findings from a cluster randomized controlled trial with 18 K-12 schools in an urban district in the US Northeast with 5878 students. Multilevel regression results, after 1 year of the intervention, showed no differences in the likelihood of suspension between students in the intervention and control schools and that the effects of the intervention did not vary by race/ethnicity, gender, or student disability status. However, for students in the intervention group, we show reductions in the likelihood of receiving an OSS for students who had previously been suspended., (© 2023. Society for Prevention Research.)

Purpose Amidst the current COVID-19 pandemic, traditional teaching methodology took a back foot. However, an urgent need for training health care worker (HCW) on preventive measures for COVID-19 infection was the need of the hour. Keeping in mind the precautionary measures required to combat COVID-19 infection, the only promising option for training was by adopting an online learning method. This study was undertaken to determine the effectiveness of video-based training using online platforms for infection prevention and control (IPC) training during the COVID-19 pandemic. Methods A quasi-experimental study, with only one experimental group comprising of HCWs, was undertaken to assess the effect of intervention which included video-assisted teaching-learning regarding IPC measures with a special focus on COVID-19 at a tertiary care Institute in North India. Online sessions were conducted on Do's and Don'ts, Hand Hygiene, donning and doffing of personal protective equipment (PPE), cleaning and disinfection, and bio-medical waste (BMW) management with the help of pre-recorded videos which was pre-validated. The session was followed by online interaction with participants by a single resource person. Pre-test and post-test were conducted through google forms before commencement and at the end of the online session respectively. The data were analyzed in terms of descriptive frequencies and percentages of different domains to describe the pre- and post-test knowledge. Results A total of 576 participants were included in the study. There was a statistically significant gain in scores of all questions related to cleaning and disinfection; and BMW. No significant change was observed in knowledge regarding the sequence of doffing (p-value: 0.94). The result of pairwise comparisons pre-test and post-test scores showed that intervention through video-assisted teaching-learning resulted in improvement of knowledge which was found to be statistically significant (p-value < 0.001). Conclusion Video-assisted teaching-learning through virtual platforms effectively trained health personnel on infection prevention and control practices during the COVID-19 pandemic. Video-assisted training can successfully be handled by a single resource person to impart the knowledge and skill to the trainee. Virtual teaching and learning is a feasible and efficient method to deliver training to HCWs on infection control practices and this methodology may be adopted in the future for several other training in manpower crunch situations, similar restrictive circumstances as posed by the COVID-19 pandemic., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2021, Sharma et al.)

The colonisation of the land by plants was accompanied by the evolution of complex tissues and multicellular structures comprising different cell types as morphological adaptations to the terrestrial environment. Here, we show that the single WIP protein in the early-diverging land plant Marchantia polymorpha L. is required for the development of the multicellular gas exchange structure: the air pore complex. This 16-cell barrel-shaped structure surrounds an opening between epidermal cells that facilitates the exchange of gases between the chamber containing the photosynthetic cells inside the plant and the air outside. Mp WIP is expressed in cells of the developing air pore complex and the morphogenesis of the complex is defective in plants with reduced Mp WIP function. The role of WIP proteins in the control of different multicellular structures in M. polymorpha and the flowering plant Arabidopsis thaliana suggests that these proteins controlled the development of multicellular structures in the common ancestor of land plants. We hypothesise that WIP genes were subsequently co-opted in the control of morphogenesis of novel multicellular structures that evolved during the diversification of land plants., (© 2017. Published by The Company of Biologists Ltd.)

Background: Anopheles mosquitoes are the only vectors of human malaria. Anopheles species use standing water as breeding sites. Human activities, like the creation of an artificial lake during the implementation of hydroelectric power plants, lead to changes in environmental characteristics and, therefore, may changes the species richness and composition of Anopheles mosquitoes. The aim of the present study was to verify whether or not there is an association between the artificial flooding resulting from the construction of the Jirau hydroelectric power plant, and the richness and composition of anophelines., Methods: Mosquitoes samples were obtained monthly from the Jirau hydroelectric power plant area located at Porto Velho, Rondônia State, using Human Landing Catch (06:00-10:00 PM). Mosquitoes collected were transported to Laboratório de Entomologia Médica FIOCRUZ-RO where they were identified until species using dichotomous key., Results: A total of 6347 anophelines belonging to eight different species were collected. The anophelines species richness was significantly lower during the first flooding stage. Differences in anophelines species composition were found when comparing the first flooding stage with the other stages. Furthermore, the mean number of Anopheles darlingi, the main vector of malaria in the region, increases during the first and the third flooding stages., Conclusions: The continual monitoring of these vectors during the late operational phase may be useful in order to understand how anophelines will behave in this area.

Purpose: Pituitary macroadenomas (MACs) represent 10-30 % of Cushing's disease (CD) cases. The aim of this study was to report the clinical, laboratorial and imaging features and postsurgical outcomes of microadenoma (MIC) and MAC patients., Methods: Retrospective study with 317 CD patients (median 32 years old, range 9-71 years) admitted between 1990 and 2014, 74 (23.3 %) of whom had MAC., Results: Hirsutism, plethora facial, muscular weakness and muscular atrophy were more frequent in the MIC patients. Nephrolithiasis, osteopenia, hyperprolactinaemia and galactorrhoea were more prevalent in MAC patients. The morning serum cortisol (Fs), nocturnal salivary cortisol (NSC), nocturnal Fs (Fs 2400 h), low- and high-dose dexamethasone suppression test results and CRH and desmopressin test results were similar between the subgroups. MIC patients showed higher urinary cortisol at 24 h (UC), and MAC patients presented higher ACTH levels but lower Fs/ACTH, Fs 2400 h/ACTH, NSC/ACTH and UC/ACTH ratios. There were negative correlations of tumour size with Fs/ACTH, Fs 2400 h/ACTH, NSC/ACTH and UC/ACTH ratios. Overall, the postsurgical remission and recurrence rates were similar between MIC and MAC. However, patients in remission (MIC + MAC) showed smaller tumour diameters and a lower prevalence of invasion and extension on MRI., Conclusions: Despite exhibiting higher plasma ACTH levels, CD patients with MAC presented lower cortisol/ACTH ratios than did patients with MIC, with a negative correlation between tumour size and cortisol/ACTH ratios. The overall postsurgical remission and recurrence rates were similar between MIC and MAC patients, with those with larger and/or invasive tumours showing a lower remission rate.

Overconsumption of fructose increases the risk of nonalcoholic fatty liver disease (NAFLD), obesity and metabolic syndrome. NAFLD is currently one of the most common etiologies of chronic liver disease worldwide. The aim of the present study is to evaluate the hepatoprotective potential of zingerone against fructose-enriched diet-induced rat model of nonalcoholic fatty liver disease. Male albino Wistar rats were used and randomly divided into four groups: group 1, control rats fed with standard pellet; group 2, rats were fed normal pellet with intragastric intubation of zingerone (100 mg/kg/day); group 3, rats were fed fructose enriched diet alone; group 4, rats were fed fructose enriched diet with intragastric intubation of zingerone (100 mg/kg/day). Body weight, abdominal circumference, blood glucose, lipid profile and hepatic function indicators were increased and HDL reduced in group 3 rats. Liver pathology of group 3 showed marked changes which includes micro- and macrovesicular steatosis, marked inflammatory cell infiltration, sinusoidal fibrosis and with a significant increase in the area percentage of the collagen. Administration of zingerone reversed the fructose enriched diet induced changes especially body weight, abdominal circumference, blood glucose, lipid profile, hepatic function indicators and restored pathological alteration of liver. Taken together these data provide new insights into the preventive approach of zingerone against the development of the NAFLD.

The partial substitution of sodium chloride by potassium chloride (0%, 25%, and 50%) and addition of arginine (1% w/w) in probiotic Minas cheese was investigated. Microbiological (Lactococcus lactis and Lactobacillus acidophilus counts, and functionality of the prebiotics L. acidophilus), physicochemical (pH, proteolysis, organic acids, fatty acids, and volatile profiles), rheological (uniaxial compression) and sensory (hedonic test with 100 consumers) characterizations were carried out. The sodium reduction and addition of arginine did not constitute a hurdle to lactic and probiotic bacteria survival, with presented values of about 9 log CFU/g, ranging from 7.11 to 9.21 log CFU/g, respectively. In addition, lower pH values, higher proteolysis, and a decrease in toughness, elasticity and firmness were observed, as well as an increase in lactic, citric, and acetic acid contents. In contrast, no change was observed in the fatty acid profile. With respect to the sensory acceptance, the probiotic low-sodium Minas cheese presented scores above 6.00 (liked slightly) for the attributes flavor and overall acceptance. The addition of arginine can be a potential alternative for the development of probiotic dairy products with reduced sodium content., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The colonization of the land by plants, sometime before 470 million years ago, was accompanied by the evolution tissue systems [1-3]. Specialized structures with diverse functions-from nutrient acquisition to reproduction-derived from single cells in the outermost layer (epidermis) were important sources of morphological innovation at this time [2, 4, 5]. In extant plants, these structures may be unicellular extensions, such as root hairs or rhizoids [6-9], or multicellular structures, such as asexual propagules or secretory hairs (papillae) [10-12]. Here, we show that a ROOTHAIR DEFECTIVE SIX-LIKE (RSL) class I basic helix-loop-helix transcription factor positively regulates the development of the unicellular and multicellular structures that develop from individual cells that expand out of the epidermal plane of the liverwort Marchantia polymorpha; mutants that lack MpRSL1 function do not develop rhizoids, slime papillae, mucilage papillae, or gemmae. Furthermore, we discovered that RSL class I genes are also required for the development of multicellular axillary hairs on the gametophyte of the moss Physcomitrella patens. Because class I RSL proteins also control the development of rhizoids in mosses and root hairs in angiosperms [13, 14], these data demonstrate that the function of RSL class I genes was to control the development of structures derived from single epidermal cells in the common ancestor of the land plants. Class I RSL genes therefore controlled the generation of adaptive morphological diversity as plants colonized the land from the water., (Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.)

In communities with an increased prevalence of hereditary deafness, social, and linguistic adaptations are found in response. Aulbers (1959) describes a high prevalence of deafness in a fishing village on the Dutch coast: Katwijk aan Zee. This article aims to assess the current prevalence of deafness in Katwijk, as well as the current sign language situation there. To this end, data were collected from various sources, including governmental studies on public health, archives, a genealogical database and interviews with deaf inhabitants of Katwijk. The various types of data confirm the presence of a higher prevalence of deafness in Katwijk that continues to date. Linguistic and anthropological research is needed to establish to what extent this has affected the experience and position of deaf people and their sign language usage in Katwijk., (© The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Objective: The expression of transcription factors involved in early pituitary development, such as PROP1 and POU1F1, has been detected in pituitary adenoma tissues. In this study, we sought to characterize the transcriptional profiles of PROP1, POU1F1, and TBX19 in functioning and nonfunctioning pituitary adenomas in an attempt to identify their roles in tumorigenesis and hormone hypersecretion., Methods: RT-qPCR analyses were performed to assess the transcriptional pattern of PROP1, POU1F1, TBX19, and hormone-producing genes in tissue samples of corticotrophinomas (n=10), somatotrophinomas (n=8), and nonfunctioning adenomas (n=6)., Results: Compared with normal pituitary tissue, POU1F1 was overexpressed in somatotrophinomas by 3-fold. PROP1 expression was 18-fold higher in corticotrophinomas, 10-fold higher in somatotrophinomas, and 3-fold higher in nonfunctioning adenomas. TBX19 expression was 27-fold higher in corticotrophinomas. Additionally, the level of TBX19 mRNA positively correlated with that of pro-opiomelanocortin (r=0.49, p=0.014)., Conclusions: Our data demonstrate that PROP1 is overexpressed in pituitary adenomas, mainly in corticotrophinomas. Together with previously published data showing that patients who harbor PROP1 loss-of-function mutations present a progressive decline in corticotrope function, our results support a role for PROP1 in pituitary tumor development and in the maintenance of cell lineages committed to corticotrophic differentiation.

The current report describes an outbreak of vesicular disease affecting dairy cows in midwestern Brazil in which a coinfection with 2 poxviruses-Vaccinia virus (VACV) and a parapoxvirus-was demonstrated. Milking cows presented vesicles, painful reddish or whitish papules, and scabby proliferative lesions in the teats and udder, in a clinical course of approximately 10-21 days. Histologically, multifocal areas of moderate to severe acanthosis, spongiosis, hypergranulosis, and parakeratotic or orthokeratotic hyperkeratosis with adjacent focally extensive ulcers were observed in the epidermis. Rounded eosinophilic inclusion bodies were observed in the cytoplasm of epithelial cells of areas with acanthosis or necrosis. Moderate inflammatory infiltrate of lymphocytes, plasma cells, neutrophils, and macrophages were observed in some dermal areas. Two people milking the affected cows developed lesions on the hands, painful papules which progressed to ulcerative and scabby lesions in 4-7 days. Electron microscopy of scabs from 1 cow revealed the concomitant presence of orthopoxvirus and parapoxvirus particles. Scabs from 2 cows were positive by polymerase chain reaction for the parapoxvirus B2L gene; 1 of the scabs was also positive for the VACV vgf gene. Nucleotide sequencing of the B2L amplicon revealed a similarity of 96-99% with Orf virus (ORFV) and lower identity with Pseudocowpox virus (92-95%) and Bovine papular stomatitis virus (85-86%). Nucleotide sequencing of a region of parapoxvirus DNA polymerase gene revealed a high similarity (98-100%) with ORFV sequences. Thus, an unusual coinfection with VACV and a parapoxvirus, likely ORFV, was demonstrated in the outbreak.

Background: Almost all land plants develop tip-growing filamentous cells at the interface between the plant and substrate (the soil). Root hairs form on the surface of roots of sporophytes (the multicellular diploid phase of the life cycle) in vascular plants. Rhizoids develop on the free-living gametophytes of vascular and non-vascular plants and on both gametophytes and sporophytes of the extinct rhyniophytes. Extant lycophytes (clubmosses and quillworts) and monilophytes (ferns and horsetails) develop both free-living gametophytes and free-living sporophytes. These gametophytes and sporophytes grow in close contact with the soil and develop rhizoids and root hairs, respectively., Scope: Here we review the development and function of rhizoids and root hairs in extant groups of land plants. Root hairs are important for the uptake of nutrients with limited mobility in the soil such as phosphate. Rhizoids have a variety of functions including water transport and adhesion to surfaces in some mosses and liverworts., Conclusions: A similar gene regulatory network controls the development of rhizoids in moss gametophytes and root hairs on the roots of vascular plant sporophytes. It is likely that this gene regulatory network first operated in the gametophyte of the earliest land plants. We propose that later it functioned in sporophytes as the diploid phase evolved a free-living habit and developed an interface with the soil. This transference of gene function from gametophyte to sporophyte could provide a mechanism that, at least in part, explains the increase in morphological diversity of sporophytes that occurred during the radiation of land plants in the Devonian Period.

Bovine papular stomatitis virus (BPSV) is a parapoxvirus associated with papular and erosive lesions on the muzzle, lips, and oral mucosa of cattle. Teats of milking cows are occasionally affected, and the infection is frequently transmitted to human beings. The present report describes an outbreak of BPSV infection affecting cows in midwestern Brazil, with human involvement. The disease was observed in neighboring small hand-milking farms, affecting 20 milking cows. The signs included painful reddish papules, ulcers, and scabby proliferative lesions on the teats, with a clinical course of 7-12 days. Affected cows presented severe local pain, not allowing the completion of milking. Histologically, acanthosis, spongiosis, and parakeratotic hyperkeratosis with adjacent focally extensive ulcers and multifocal inflammatory infiltrate were observed in the epidermis. Eosinophilic inclusion bodies were noted in the cytoplasm of epithelial cells. Personnel milking the affected cows developed lesions on the hands, painful papules that progressed to ulcerative and scabby lesions in 4-7 days. A polymerase chain reaction using a set of pan-parapoxvirus primers for the B2L gene performed on DNA extracted from scabs amplified a 590-bp product, which when sequenced, revealed similarities of 99%, 85%, and 84% with BPSV, Pseudocowpox virus, and Orf virus, respectively. A phylogenetic tree based on the B2L sequence was constructed, showing that the virus clustered with BPSV isolates. Although clinical cases compatible with BSPV infection have been frequently described in Brazil, the present report identifies the agent associated with cattle and human disease in the country.

Diffuse large B-cell lymphoma (DLBCL) is a heterogeneous disease. Certain chromosomal translocations are associated with clinical outcome, but it is likely that there are both tumor suppressor genes and oncogenes that cooperate with the primary translocations. We have used the Mitelman database to compare chromosomal losses and gains of DLBCL possessing t(14;18), t(8;14), or t(3;14) with DLBCL lacking any of these translocations. The data we obtained are low resolution, but results for t(3;14) validate the methodology. In accord with the literature, loss of 6q was associated with t(3;14). Chromosomes 11, 13, and X were gained significantly in t(3;14), whereas 8p23 was lost. Cases with t(14;18) were associated with gains of chromosomes 7 and 12; cases with t(8;14) were associated with gains of chromosomes 1 and 4.

Purpose: To evaluate the ability of polymeric nanocapsules (NCs) radiolabeled with technetium-99m D,L-hexamethylpropyleneamine oxime (Tc-HMPAO) to identify inflammatory process in an experimental model., Methods: NCs were prepared by interfacial deposition of preformed biodegradable polymer [poly (D,L-lactic acid) (PLA) and PLA-PEG (polyethyleneglycol)] followed by a solvent displacement. The size and homogeneity, and zeta potential of the NC preparations were determined in a Zetasizer by photon correlation spectroscopy and laser Doppler anemometry, respectively. The NCs radiolabeled with Tc-HMPAO were administered intravenously to Wistar male rats bearing a focal inflammation induced by subplantar injection of carrageenan in the right foot. At preestablished time intervals, the animals were anesthetized, tissues were removed and radioactivity was determined using an automatic scintillation apparatus., Results: The average diameter calculated by photon correlation spectroscopy varied from 216 to 323 nm. The biodistribution studies showed a greater uptake of the PEG surface-modified Tc-HMPAO-NC by the inflamed paws when compared with the respective controls. There was no significant difference in the uptake of conventional Tc-HMPAO-NC and of free Tc-HMPAO by inflamed and control paws. These results indicate that the PLA-PEG Tc-NC showed a higher uptake in inflammation compared with free complex and may be useful as a radiotracer to identify these foci.

Unlabelled: In the early postoperative period of Cushing's disease patients, desmopressin may stimulate ACTH secretion in the remnant corticotrophic tumour, but not in nontumour suppressed cells., Objective: The aim of this study is to evaluate the serum cortisol responses to desmopressin after pituitary surgery, establishing an optimal cut-off for absolute increment (Delta) of serum cortisol (F) suitable to predict recurrence risk., Design: Retrospective case record study., Patients: Fifty-seven Cushing's disease patients submitted to pituitary surgery and desmopressin stimulation in the early postoperative with a long-term follow-up (20-161 months) were studied., Methods and Measurements: Serum cortisol levels after desmopressin test (10 microg i.v.) 15-30 days after adenomectomy were used to determine DeltaF (absolute increment of F: F peak - F baseline). Sensitivity and specificity of DeltaF were calculated and a ROC curve was performed to establish an optimal cut-off for DeltaF to predict recurrence risk., Results: Fifteen patients had immediate postoperative failure (basal F > 165 nmol/l; 6 microg/dl) and one patient was lost during the follow-up. Forty-one patients achieved initial remission and were followed-up. Five of 11 patients who recurred had DeltaF > 193 nmol/l (7 microg/dl), but none of 30 patients who remained in prolonged remission showed DeltaF > 193 nmol/l after postoperative desmopressin stimulation., Conclusions: Persistence of cortisol response (DeltaF > 193 nmol/l) to desmopressin in the early postoperative period can help to identify Cushing's disease patients with initial remission who present risk for later recurrence.

Objective: GH secretagogues (GHS) produce exaggerated ACTH and cortisol responses in Cushing's disease (CD) patients, attributable to their direct action on GH-releasing peptide receptor type 1a (GHSR-1a). However, there are no studies correlating the in vivo response to GHS and GHSR-1a mRNA expression in ACTH-dependent Cushing's syndrome (CS) patients. The aim of this study is to correlate the patterns of ACTH and cortisol response to GH-releasing peptide-6 (GHRP-6) to GHSR-1a expression in ACTH-dependent CS patients., Design: Prospective study in a tertiary referral hospital center. Fifteen CD patients and two ectopic ACTH syndrome (EAS) patients were studied., Methods: Tumor fragments were submitted to RNA extraction, and GHSR-1a expression was studied through real-time qPCR and compared with normal tissue samples. The patients were also submitted to desmopressin test and vasopressin receptor type 1B (AVPR1B) mRNA analysis by qPCR., Results: GHSR-1a expression was similar in normal pituitary samples and in corticotrophic tumor samples. GHSR-1a expression was higher in patients (CD and EAS) presenting in vivo response to GHRP-6. Higher expression of AVPR1B was observed in the EAS patients responsive to desmopressin, as well as in corticotrophic tumors, as compared with normal pituitary samples, but no correlation between AVPR1B expression and response to desmopressin was observed in the CD patients., Conclusions: Our results revealed a higher expression of GHSR-1a in the ACTH-dependent CS patients responsive to GHRP-6, suggesting an association between receptor gene expression and in vivo response to the secretagogue in both the CD and the EAS patients.

Deletions at the t(9;22) breakpoint regions, found in 15% of chronic myeloid leukemia patients (CML) with an overt Philadelphia (Ph) translocation, are associated with an adverse disease prognosis in patients receiving interferon-alpha therapy. The incidence of deletions has been shown to vary for different cytogenetic subgroups of CML, with a significantly higher incidence of deletion in patients with a variant Ph translocation. To date, however, the frequency of such deletions in the subgroup of CML patients in whom the BCR/ABL1 fusion arises via submicroscopic chromosomal insertion (masked Ph) has not been investigated. We report the evaluation of 14 patients with masked Ph-positive CML for the presence of deletions extending 3' from BCR and 5' from ABL1 using two triple-color BCR/ABL probes. Deletions were identified in 3 patients (21%), encompassing sequences 5' to ABL1 in two of these and sequences 3' to BCR in the remaining patient, thus demonstrating that the phenomenon is a significant feature of the masked Ph CML subgroup. Furthermore, our findings are consistent with the notion that loss of genomic material is a potential side effect of any DNA breakage event at the 9q34.1 and 22q11.2 chromosomal regions, regardless of the subsequent mechanism of chromosomal rearrangement.

Complement activation is an important step in the acceleration of liposome clearance. The anaphylatoxins released following complement activation may motivate a wide variety of physiologic changes. We performed physicochemical characterization and in vitro studies of the interaction of complement system with both noncirculating and long-circulating pH-sensitive and nonpH-sensitive liposomes. The liposomes were characterized by diameter, zeta potential, and atomic force microscopy (AFM). The study of liposome interactions with complement system was conducted using hemolytic assay in rat serum. All liposomes presented a similar mean diameter (between 99.8 and 124.3 nm). The zeta potential was negative in all liposome preparations, except in liposomes modified with aminopoly (ethyleneglycol) 2000-distearoylphosphatidylethanolamine (aPEG(2000)-DSPE), which presented positive zeta potential. Atomic force microscopy images showed that non-long-circulating pH-sensitive liposomes are prone to vesicles aggregation. Non-pH-sensitive liposomes complement system activates, while pH-sensitive liposomes showed to be poor complement activators in rat serum.

Purpose: To investigate the biodistribution and the ability of stealth pH-sensitive liposomes radiolabelled with 99mTc to identify inflammatory regions in a rat focal inflammation model., Methods: Preformed glutathione-containing stealth pH-sensitive liposomes were labelled with 99mTc-hexamethylpropylene amine oxime (99mTc-HMPAO). The 99mTc-HMPAO radiolabelled stealth pH-sensitive liposomes (99mTc-SpHL) were administered intravenously in Wistar male rats with inflammation induced by injection subplantar of carrageenan in the right foot. At pre-established time intervals the animals were anaesthetized and tissues were removed and analysed for 99mTc content using an automatic scintillation apparatus. Scintigraphic imaging was also performed after 2, 4 and 8 h of intravenous injection of 99mTc-SpHL., Results: The 99mTc-SpHL was significantly taken up by the spleen (19.21+/-2.98%ID/g at 30 min post-injection). Low radioactivity levels were found in the liver, lungs, and kidney. Moreover, the 99mTc-SpHL uptake was significantly higher in the inflamed foot when compared to the respective control (0.386+/-0.059 and 0.215+/-0.018%ID/g at 2 h post-injection, respectively). As early as 30 min after administration of 99mTc-SpHL, the focus of inflammation could be visualized scintigraphically. The value of the inflammatory and non-inflammatory site radioactivity counting ratio was greater than 5., Conclusion: This result indicates that the 99mTc-SpHL presents a high tropism for inflammatory regions and may be useful as a radiopharmaceutical to identify these foci.

Thymic hyperplasia has been described after the resolution of hypercortisolism from several etiologies, causing great diagnostic dilemmas. We describe a case where the catheterization of the thymic vein was essential for the differential diagnosis of a thymic enlargement in an adrenalectomized patient with ACTH-dependent Cushing's syndrome. The patient was a 48-year-old female with clinical and laboratorial data suggesting Cushing's disease. She underwent a transsphenoidal surgery with no tumor visualization and no remission of the syndrome. Histopathological studies disclosed a normal pituitary. She underwent a bilateral adrenalectomy and 8 months later a chest CT showed an increase of left thymic lobe, which was previously non-existent. After a negative (111)In-pentetreotide scintigraphy, the patient underwent simultaneous and bilateral catheterism of the petrosus sinuses and catheterization of the thymic and inominate veins and no ACTH gradient was shown among the sites of collection. She did not undergo thoracotomy and a follow-up was established. During the evolution, there was a spontaneous regression of the thymic lesion 38 months after the diagnosis. The ACTH gradient during the catheterization of thymic vein was essential for the differential diagnosis of the thymic enlargement tumor after hypercortisolism resolution in ACTH-dependent Cushing's syndrome, especially in this case, where the ACTH source was occult, thus avoiding an invasive surgical procedure for a benign entity with spontaneous resolution.

Numerous studies conducted in developed countries demonstrate that persons living with a with severe mental illness (SMI) are at elevated risk for HIV infection. Fewer studies have addressed this topic in the developing world, and no study has evaluated the effects of a risk reduction intervention. Because risk reduction requires adequate knowledge regarding HIV-related transmission and prevention strategies, the current study sampled patients diagnosed with SMI in India and assessed (a) knowledge regarding HIV/AIDS, and (b) short-term retention of knowledge following HIV risk reduction education. Patients were assessed for HIV knowledge at baseline, received an HIV educational program, and then were re-assessed for their knowledge one and five days later. The results indicated a poor level of baseline knowledge, which improved following education; knowledge gains were sustained at five days. Men, and patients with college education, demonstrated better knowledge. However, significant gains in knowledge were observed among all patients regardless of gender, education, psychiatric diagnosis, and prior sexual risk behavior. The findings indicate that a brief, HIV-focused educational intervention can help to improve knowledge among Indian psychiatric patients.

Mitochondrial FAS (fatty acid synthesis) of type II is a widely conserved process in eukaryotic organisms, with particular importance for respiratory competence and mitochondrial morphology maintenance in Saccharomyces cerevisiae. The recent characterization of three missing enzymes completes the pathway. Etr1p (enoyl thioester reductase) was identified via purification of the protein followed by molecular cloning. To study the link between FAS and cell respiration further, we also created a yeast strain that has FabI enoyl-ACP (acyl-carrier protein) reductase gene from Escherichia coli engineered to carry a mitochondrial targeting sequence in the genome, replacing the endogenous ETR1 gene. This strain is respiratory competent, but unlike the ETR1 wild-type strain, it is sensitive to triclosan on media containing only non-fermentable carbon source. A colony-colour-sectoring screen was applied for cloning of YHR067w/RMD12, the gene encoding mitochondrial 3-hydroxyacyl-ACP dehydratase (Htd2/Yhr067p), the last missing component of the mitochondrial FAS. Finally, Hfa1p was shown to be the mitochondrial acetyl-CoA carboxylase.

Several studies have attempted to understand the link among substance abuse, depression, and suicidal ideation (SI). Assessment of this link is important to develop specific interventions for persons in substance abuse treatment. This association was tested among 990 drug users in and out of treatment with significant criminal justice histories from two National Institute on Drug Abuse studies. The Diagnostic Interview Schedule and Substance Abuse Module assessed DSM-III-R depression, number of depression criteria met, antisocial personality disorder (ASPD), and substance use disorders. Compared with men, women were twice as likely to report depression (24% vs. 12%), whereas men were nearly twice as likely to report ASPD (42% vs. 24%). High rates of SI were found, with women more likely than men to report thoughts of death (50% vs. 31%), wanting to die (39% vs. 21%), thoughts of committing suicide (47% vs. 33%), or attempting suicide (33% vs. 11%); 63% of women and 47% of men reported at least one of these suicidal thoughts or behaviors. Male and female ideators were more likely than nonideators to report depressed mood and to meet criteria for depression, ASPD, and alcohol use disorders. Male ideators were more likely than male nonideators to meet criteria for cocaine use disorders. Using logistic regression, SI among men was predicted by alcohol use disorder (OR = 1.60), ASPD (OR = 1.59), and number of depression criteria (OR = 9.38 for five criteria). Among women, SI was predicted by older age, marital status, alcohol use disorder (OR = 2.77), and number of depression criteria (OR = 9.12 for five criteria). These original findings point out the need to discuss suicidal thoughts among depressed drug users for early treatment and prevention.

Background: The most common way to prevent transmission of CMV by blood transfusion is to use blood products from seronegative donors. Screening of blood donors for CMV infection is usually based on detection of antigens obtained from the CMV laboratory strain AD 169. Recent evidence suggests that approximately up to 20 percent of CMV-negative blood donors may in fact be CMV-DNA positive by PCR analyses., Study Design and Methods: In this study, sera from CMV-seronegative, CMV-seropositive, and CMV-DNA-positive/seronegative individuals, and from patients with acute and convalescent CMV infection for detection of CMV antibodies were analyzed. CMV antigens prepared from cells infected with CMV clinical isolates or the CMV laboratory strain AD 169 in ELISA and Western blot assays were used., Results: All CMV-positive sera from blood donors were seropositive for the CMV antigens prepared from AD 169 (A2) or from a CMV clinical isolate (C6). Interestingly, whereas all CMV-negative blood donors were negative in tests for the CMV antigen A2, 36 percent were CMV seropositive using the CMV antigen C6 in ELISA., Conclusion: The data suggest that a substantial number of CMV-seronegative/CMV-DNA-positive serum samples contain antibodies that recognize CMV clinical isolate antigens.

Background and Objectives: The acquisition of additional cytogenetic changes (clonal evolution, CE) during treatment of chronic myeloid leukemia (CML) with imatinib mesylate is currently regarded as an index of increasing resistance to imatinib. Therefore, to investigate whether CE as an isolated event increases the risk of disease progression during imatinib treatment, we compared the outcome of patients with CML in chronic phase (CML-CP) who developed CE whilst in complete hematologic remission with the outcome of comparable patients in complete hematologic remission who showed no evidence of CE., Design and Methods: We serially studied cytogenetic findings in 102 patients receiving the Abl-tyrosine kinase inhibitor, imatinib mesylate, as sole agent to treat CML-CP who had no evidence of CE before initiation of imatinib treatment., Results: CE was identified during treatment with imatinib in 15 patients, 10 of whom were in complete hematologic remission. In most cases these changes occurred exclusively in the Ph+ population but in three patients additional changes occurred in a co-existing Ph-negative population. Patients with de novo CE in the absence of any other sign of disease progression had a significantly higher incidence of progression by 18 months than did non-CE patients (progression-free survival 34.3% (CI 10.5-69.8%) vs. 94.1% (CI 80.6-98.4%), p<0.0001)., Interpretation and Conclusions: Based on this relatively small series of patients, we conclude that acquisition of clonal evolution increases the risk of subsequent disease progression also in CML patients in complete hematologic remission on imatinib.

Background & Objectives: An association has been shown between alcohol use and high-risk sexual behaviour. One of the important variables that might mediate risky sexual behaviour among alcohol users is the sensation-seeking dimension of personality. This study was undertaken with the aim to assess the relationship between sensation seeking and sexual risk taking among heavy alcohol users admitted to a De-addiction centre., Methods: To study the above association, sensation seeking was compared between subjects admitted for treatment of alcohol dependence with (n = 30) and without (n = 30) reported high-risk sexual behaviour., Results: Subjects reporting high-risk sexual behaviour were found to have higher sensation seeking scores in all the domains of the sensation seeking scale compared to those without high-risk sexual behaviour. Scores on the subscale disinhibition, of the sensation seeking scale seemed to be associated with higher sexual risk taking., Interpretation & Conclusion: Sensation seeking as a personality variable was significantly associated with sexual risk taking behaviour among heavy alcohol users. The study indicates the need for careful assessment of personality in these individuals and emphasizes the need for further studies on a larger sample.

Many pharmacological and surgical treatments are available for the treatment of chronic facial pain. However, many of them are expensive and often very ineffective. Past publications suggested that bulbar trigeminal stereotactic nucleotractotomy is a very useful procedure for the treatment of neuropathic or oncologic facial pain. The authors describe the results of treatment with stereotactic nucleotractotomy in 58 patients with chronic facial pain. The intensity of the pain was evaluated according to the visual analogue scale, and daily life activities were also evaluated. The conclusion was that this procedure is a safe and effective method for treatment of postherpetic neuralgia, Wallenberg's syndrome and oncologic facial pain but not of trigeminal neuralgia., (Copyright 2003 S. Karger AG, Basel)

The mutagenic activity of glycine upon nitrosation was studied in the Ames tester strains TA98, TA100, TA102, and TA104. The results obtained show that glycine at acidic pH values and in the presence of Cl- can react with nitrite giving rise to genotoxic compounds to the tester strains used. When these experiments were carried out in the presence of gastric juice the genotoxicity observed was associated with the Cl- concentration in the different gastric juice samples. The nature and the mechanism of genetic lesion induced by the ultimate genotoxicant arising from the nitrosation of glycine are not fully understood. Primary amines (e.g., amino acids) have been described as potential alkylating agents after nitrosation. However, in our experimental conditions these alkylating activities were not detected, suggesting that other mechanisms could be involved in the genetic lesion induced by nitrosated glycine. The influence of Cl- in the genotoxic activity of glycine and other primary amines upon nitrosation and its possible involvement in the etiology of gastric cancer are discussed.

We conducted a prospective clinical trial of 154 patients with duodenal ulcer disease, which was divided in two different steps: healing phase and post-healing phase. In the first phase, the patients were randomly assigned to treatment with H2 blockers (Ranitidine 300 mg or Famotidine 40 mg), in a single bedtime dose. Endoscopy was performed after 4 and 12 weeks of treatment and the healing rate was 70% and 90%. There were no significant differences between the two drugs. After 12 weeks of treatment there were sixteen patients with non-healed duodenal ulcer, considered as refractory duodenal ulcer disease, in which we started therapy with a double dose of H2 receptor antagonists or sucralfate (4 gr/day). Endoscopic follow-up after 12 weeks of this regimen revealed ulcer healing in about half of them; the cummulative healing rate in 6 months was 96.4%. Persistent lesions after 6 months of follow-up were found in 5 patients. Omeprazole 40 mg, single dose, was given and all of them were healed during the first 4 weeks of treatment. In the second phase, 137 patients healed during the first 3 months of treatment, were followed for 12 months and randomized in two groups: 61, with maintenance therapy--ranitidine 150 mg or famotidine 20 mg; 76, without treatment. In the patients without maintenance therapy, the recurrence rate was significantly higher (p less than 0.55), than in the group with maintenance therapy. In this group several parameters were evaluated--clinical, biochemical and psychosocial--to try to identify some prognostic markers of healing, relapse.(ABSTRACT TRUNCATED AT 250 WORDS)