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Objective: This study was carried out to investigate the possible effect of Ficus vogelii (FV) on liver function and some he matological parameters of male Wistar rats administered with aspirin. Materials and Methods: The animals were divided into 4 groups comprisingx 5 animals each. Group 1: Normal control group, Group 2: Positive control group which received 500 mg/kg of aspirin for three days. Group 3: Pre-treated with aqueous extract of Ficus vogelii (600 mg/kg body weight-bwt) for 14 days and then administered with Aspirin (500 mg/kg bwt) for 3 days. Group 4: received 500mg/kg of aspirin for three days, and then treated with aqueous extract of FV (600 mg/kg bwt) for 14 days. Results: There was a significant decrease (p<0.05) in PCV in the treatment groups 2, 3 and 4 (39.00±5.19, 41.50±2.59 and 41.50±1.91% respectively) when compared to the control group (62.50±1.44%). A significant decrease was recorded in WBC and platelet count in group 3 (pre-treatment) and 4 (treatment groups) (9.6±0.40×10/L and 8.8±1.10 × 10/L respectively) when compared to group 2. A significant decrease was also observed in AST, ALT and ALP levels in FV treated group 4 (9.50±0.29 and 138.50±3.75 μ/L respectively) when compared to group 2 (aspirin-treated group: 27.50±0.87 and 194.50±12.41 μ/L respectively). Conclusion: This extract may thus be useful in ameliorating Aspirin induced liver damage.

Introduction: Parkinson’s disease is a second most common neurodegenerative disorder after Alzheimer’s disease. It affects 1% of population over age of 50 years. Non-Motor symptoms are common in patients of Parkinson’s disease. These remain undiagnosed most of the time. Aim and Objectives: To analyse gender specific, age related and demographic variability of non-motor symptoms (Cardiovascular symptoms, sleep disorder, hallucination, sexual disorders, apathy, pain and miscellaneous symptoms) of Parkinson’s disease. Materials and Methods: Present study included 100 cases (66 males and 34 females) of Parkinson’s disease presenting at department of neurology, Dayanand Medical College and Hospital, Ludhiana. 30 questionnaires of non-motor symptoms in Hindi and Punjabi were given to patients. Gender, age, duration of non-motor symptoms, Parkinson’s phenotype and complete demographic profile of each patient taken into account for study. Thereafter result were statistically analysed for all parameters. Conclusion: It is concluded that mean age and duration of non-motor symptoms were higher in males as compared to females. It was calculated that there was positive correlation of H&Y with age and non-motor symptoms like Cardiovascular, Sleep related disorder, apathy, pain and miscellaneous symptoms. As H&Y increases there is increase in frequency and severity of symptoms. There was difference in dominant non-motor symptoms among different genders. Sexual dysfunction and sleep disorders were more prominent in males whereas cardiovascular and miscellaneous like excessive sweating were more in females.

Chromium (Cr), an essential trace element, turns into an endocrine disruptor and male reproductive toxicant when its concentration in drinking water exceeds the safe limit. Improper disposal of effluents from more than 50 industries that use Cr contaminate the environment, in addition to occupational exposure of the workers. Testis has come to stay as a target for the reproductive toxicity of hexavalent Cr (CrVI), whereas its impact on fetal testicular differentiation remains elusive. We tested the hypothesis “In utero exposure to CrVI may alter the level of specific proteins controlling differentiation of testicular cell types”. Pregnant Wistar rats were exposed to drinking water containing 50, 100 and 200 ppm potassium dichromate (CrVI) during gestational days 14 to 21, covering the period of fetal differentiation of testicular cells. Testes were collected on postnatal day 1 and subjected to light microscopic histological studies and immunohistochemical detection of cell-specific proteins. Testis of neonatal rats with gestational exposure to high doses of CrVI showed shrunken and dispersed tubules with fewer gonocytes, extensive vacuolization of seminiferous cord accompanied by damaged epithelium, and shrunken Leydig cells present in large interstitial spaces and loose compaction of cells when compared coeval control group. Immunosignals of androgen and estrogen receptor β increased, whereas those of estrogen receptor α, follicle stimulating hormone receptor, anti-Mullerian hormone, P450 aromatase, inhibin, c-fos and c-jun decreased. Immunosignals of steroidogenic acute regulatory protein and CYP11A1 increased, whereas 3β - hydroxy steroid dehydrogenase and CYP17A1 proteins decreased, indicating compromised steroidogenic function. Our findings support the proposed hypothesis and we conclude that gestational exposure to CrVI disrupts specific hormones and hormone receptors that control fetal differentiation of testicular cells. The detrimental effect of gest

The thalamo-cortical circuit is important in the genesis of absence epilepsy. This circuit can be influenced by connecting pathways from various parts of central nervous system. The aim of the present study is to define the dento-thalamic connections in Wistar animals and compare the results with genetic absence epilepsy rats from Strasbourg (GAERS) using the biotinylated dextran amine (BDA) tracer. We injected BDA into the dentate nucleus of 13 (n=6 Wistar and n=7 GAERS) animals. The dento-thalamic connections in the Wistar animals were denser and were connected to a wider range of thalamic nuclei compared with GAERS. The dentate nucleus was bilaterally connected to the central (central medial [CM], paracentral [PC]), ventral (ventral medial [VM], ventral lateral [VL], and ventral posterior lateral [VPL]), and posterior (Po) thalamic nuclei in Wistar animals. The majority of these connections were dense contralaterally and scarce ipsilaterally. Contralateral connections were present with the parafascicular (PF), ventral posterior medial, ventral anterior (VA), and central lateral (CL) thalamic nuclei in Wistar animals. Whereas in GAERS, bilateral connections were observed with the VL and CM. Contralateral connections were present with the PC, VM, VA, and PF thalamic nuclei in GAERS. The CL, VPL, and Po thalamic nucleus connections were not observed in GAERS. The present study showed weak/deficit dento-thalamic connections in GAERS compared with control Wistar animals. The scarce information flow from the dentate nucleus to thalamus in GAERS may have a deficient modulatory role on the thalamus and thus may affect modulation of the thalamo-cortical circuit., NA

Introduction: Incidence of breast cancer is high and constitutes a significant cause of female mortality. One of breast cancer type is Estrogen and Progesteron Receptor (ER and PR) positive, therefore affected by estrogen and progesteron concentration. Treatment of Thyponium flageliforme in combination with Curcuma zedoaria, and Phyllanthus niruri (TCP) have been proven capable of inhibiting tumor cells proliferation. Objective: To evaluate the expression of protein estrogen receptor and caspase-9 in C3H mice were inoculated with adenocarcinoma cells. Methods: 18 mice were inoculated with one million cells of breast adenocarcinoma per mouse and assigned into three groups. TCP with dosage 85 and 125 mg were treated for 21 days to evaluate the expression of protein estrogen receptor and caspase-9 measured by immuno-histochemical staining. Results: Mann Whitney analysis demonstrated that expression of protein estrogen receptor and caspase-9 in TCP 125 was significant (p < 0.05). Conclusion: Expression of protein estrogen receptor was decreased and caspase-9 expression was increased by TCP administration.

Introduction: People worldwide drink Zamzam water either medicinally or religiously. However, the effect of zamzam water extract of CZ in breast cancer cells remains unclear. Objective: To evaluate the zamzam water extract of CZ effect on cell apoptosis, p53 and Bcl2 protein expression in T47 D cells line culture. Methods: In the control group design, 16 Wells which was filled with T47 D cell line culture in the density of 5 × 104/100 μl, were assigned into 4 groups. Control group (C-G), were only filled with T47D cell line culture; Zamzam 50 (ZZ-50); Zamzam 100 (ZZ-100), and Zamzam 500 (ZZ-500) groups, were filled with T47D cell line cultures and 50, 100, and 500 μg/ml zamzam water extract of CZ respectively, and then incubated for 48 hours. The number of apoptotic cell was counted by flow cytometry, while p53 and Bcl2 measured by immunohistochemistry staining method. Results: Mann Whitney analysis indicated that the percentage of apoptotic cell and p53 protein expression in ZZ-100 was significant higher compared to that of C-G (p < 0.05). In contrary the percentage number of Bcl2 protein expression in ZZ100-G was significantly lower when compared to that of C-G (p < 0.05). Conclusion: Treatment of zamzam water extract of CZ on T47D cell line cultured was capable of increasing the number of apoptotic cells, p53 protein expression, but decreased Bcl2 protein expression in T47D cell line culture. However the numbers of apoptotic cells were less than 10%.

The role of cerebellum in coordination of somatic motor activity has been studied in detailed in various species. However, experimental and clinical studies have shown the involvement of the cerebellum with various visceral and cognitive functions via its vast connections with the central nervous system. The present study aims to define the cortical and subcortical and brain stem connections of the cerebellum via the superior (SCP) and middle (MCP) cerebellar peduncle using biotinylated dextran amine (BDA) and Fluoro-Gold (FG) tracer in Wistar albino rats. 14 male albino rats received 20-50-n1 pressure injections of either FG or BDA tracer into the SCP and MCP. Following 7-10 days of survival period, the animals were processed according to the related protocol for two tracers. Labelled cells and axons were documented using light and fluorescence microscope. The SCP connects cerebellum to the insular and infralimbic cortices whereas, MCP addition to the insular cortex, it also connects cerebellum to the rhinal, primary sensory, piriform and auditory cortices. Both SCP and MCP connected the cerebellum to the ventral, lateral, posterior and central, thalamic nuclei. Additionally, SCP also connects parafasicular thalamic nucleus to the cerebellum. The SCP connects cerebellum to basal ganglia (ventral pallidum and clastrum) and limbic structures (amygdaloidal nuclei and bed nucleus of stria terminalis), however, the MCP have no connections with basal ganglia or limbic structures. Both the SCP and MCP densely connects cerebellum to various brainstem structures. Attaining the knowledge of the connections of the SCP and MCP is important for the diagnosis of lesions in the MCP and SCP and would deepen current understanding of the neuronal circuit of various diseases or lesions involving the SCP and MCP., NA

Cynodon dactylon has been mentioned in ayurvedic system of Indian medicine to be of value in the treatment of diabetes mellitus. The purpose of this study was to investigate the possible hypoglycemic effects of aqueous extract of the whole plant of Cynodon dactylon on blood glucose, plasma insulin, serum lipid profiles and possible histopathological changes in the pancreatic tissue of streptozotocin induced diabetic rats. Type 1 diabetes was induced by streptozotocin (STZ) in rats. One sub-group of rats, received glibenclamide treatment and another received Cynodon extract Treatment for 45 days respectively. Blood was analyzed for fasting blood glucose, insulin levels and lipid profiles after treatment. Histopathological sectional studies of the pancreatic islets were done. Treatment of the animals with the aqueous extract from plant at a dose of (500 mg/kg body weight) for 45 days resulted in significant reduction in blood glucose, cholesterol, triglycerides, low-density lipoprotein, and very- low - density lipoprotein in STZ diabetic rats. These biochemical parameters were correlated with the histopathological changes observed in the pancreatic islets of STZ diabetic rats, which structurally proved the effectiveness of the aqueous extract of C.dactylon in STZ diabetic rats. The obtained biochemical and histopathological results confirmed the hypoglycemic and antihyperlipidemic effects of aqueous extract of the C.dactylon in experimental model of diabetic animals. Cynodon dactylon may be useful as a natural supplementary medicine in the treatment of diabetes and its complications.

Objectives: Asafetida is traditionally used in folklore medicine for the treatment of various ailments. To validate its use in traditional medicine, it is important to evaluate its toxicity in the animal system. Therefore, this study aimed to evaluate the toxicological effects of asafetida in Wistar albino rats. Materials and Methods: Acute toxicity tests were conducted by the oral administration of 250, 500, and 1,000 mg/kg body weight of the animal. In chronic study, animals were administered with various doses of asafetida (25, 50, 100, and 200 mg/kg body weight) for a period of 6 weeks. At end of experiment, the effects of asafetida on hematological, renal, and hepatic markers and histological parameters were analyzed. Results: In acute toxicity study, no mortality was seen up to 72 h of the administration of asafetida. No signs of neurological and behavioral changes were noticed within 24 h. In the chronic study, the asafetida intake has changed the hematological parameters such as red blood cell (RBC), white blood cell (WBC), hematocrit (HCT), and platelets. Aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) were significantly increased in treated animals. The plasma level of urea and creatinine were not altered by the administration of asafetida throughout the study. Histopathology study indicates hepatotoxicity, but no signs of prominent pathological changes in kidney. Conclusions: Asafetida did not show any acute toxicity, but chronic administration could have undesirable effects on hepatocytes and hematological factors.

Arteria comitans nervi ischiadici or Persistent sciatic artery (PSA) is a rare congenital vascular anomaly with an incidence varying 0.025-0.04 % . During routine dissection in department of anatomy an incomplete type of Persistent sciatic artery was found which is being reported here. Its oncogeny and phylogeny is also discussed in detail. Such a PSA is prone to atheromatous degeneration, thrombosis, distal thromboembolism, aneurysm formation and rupture. Although rare, the possibility of such an anomaly must be borne in mind with certain clinical presentations , during orthopedic procedures on the hip and during angiographic studies of the leg. Successful surgical correction of problems necessitates excluding the anomalous artery from the circulation while revascularizing the lower exterimity.

Breast cancer is the most common cancer and the second leading cause of cancer death in women. Combinations of Thyponium flageliforme, Curcuma zedoaria and Phyllanthus niruri (TCP) have been proven as immunomodulator and retarded tumor progression in vivo. Objective: to investigate the apoptotic and anti-proliferative effect of TCP in C3H mice suffered from adenocarcinoma cells. Methods: 18 mice inoculated with one million cells of breast adenocarcinoma per mouse were assigned into three groups, TCP with dosage 85 and 125 mg were treated for 21 days to evaluate tumor volume, expression of Ki-67 and Capase3. Expression of Ki-67 and Capase3 were measured by immunohistochemistry staining, whilst cancer volume was assessed using calipers. Results: Mann Whitney analysis indicated that tumor volume in TCP groups were lower, otherwise for expression of caspase3, p < 0.05 respectively. Whereas expression of Ki-67 in TCP groups was lower, but not significant, p > 0.05. Conclusion: TCP treatment with 85 and 125 mg dosage capable of reducing tumor volume, increasing caspase3 and lowering Ki-67 expression.

Although minutes of a spinning episode may induce vertigo in the healthy human, as a result of a possible perceptional plasticity, Sufi Whirling Dervishes (SWDs) can spin continuously for an hour without a vertigo perception.This unique long term vestibular system stimulation presents a potential human model to clarify the cortical networks underlying the resistance against vertigo. This study, therefore, aimed to investigate the potential structural cortical plasticity in SWDs. Magnetic resonance imaging (MRI) of 10 SWDs and 10 controls were obtained, using a 3T scanner. Cortical thickness in the whole cortex was calculated. Results demonstrated significantly thinner cortical areas for SWD subjects compared with the control group in the hubs of the default mode network (DMN), as well as in the motion perception and discrimination areas including the right dorsolateral prefrontal cortex (DLPFC), the right lingual gyrus and the left visual area 5 (V5)/middle temporal (MT) and the left fusiform gyrus. In conclusion, this is the first report that warrants the potential relationship of the motion/body perception related cortical networks and the prolonged term of whirling ability without vertigo or dizziness., YOC, School of Biomedical Sciences, University of Otago

We studied the effects of ethanolic leaf extract of Azadirachta indica (AIE) on hepatic microscopic anatomy and oxidative stress markers in diabetic rats. Seventy-five Wistar rats (8 weeks old) were randomly assigned to five treatment groups: control; diabetic; diabetic+AIE; AIE only; and diabetic+glibenclamide. Hyperglycemia was induced in fasted rats with streptozotocin. AIE was administered orally at 500 mg/kg bw/d and glibenclamide at 600 μg/kg bw/d for 50 days (50 d). Animals were sacrificed on treatment days 7, 21 and 50. The liver was stained with PAS. Hepatic markers of oxidative stress were also estimated. At 50 d, histological study of the liver of diabetic rats showed swollen PAS+ hepatocytes, whose content was confirmed to be glycogen. On the contrary, hepatocytes of AIE-treated diabetic rats lacked glycogen. The major finding in these rats was exacerbated oxidative stress. Our findings in this model showed the beneficial effect of AIE in the amelioration of diabetic hepatic glycogenosis.Keywords: Diabetes, Azadirachta indica, hepatic glycogenosis, oxidative stress, liver

Encouraging the participation of students in the learning process, the authors introduced changes to the method of group learning, which changed the presentation on two topics from the general embryology. The purpose of the study was to assess students' perceptions of this method in comparison with the lecture-form training. To do this, the method of questioning was used at the Department of Anatomy of the Indian Medical College. Materials and methods. In general, in the general course of embryology, eight sections were identified. The study of six of them was conducted using the traditional didactic method of lectures. Assimilation of the other two sections was carried out with the help of a modified method of group learning. The students were divided into five groups, each of which received a set of materials containing basic factual information, four clinical history histories and preliminary exam questions on topics. During the discussion of what happened in the presence of the teacher, students were able to evaluate the classes using questionnaires. Results Thus, most students noted that learning with the help of a modified group method facilitated the accomplishment of tasks (46 students, 85%), made it possible to better understand the teaching material (43 students, 79%), became more interesting (43 students, 81%), provided mass participation of students in the educational process (51 students, 94%) and the application of their greater efforts (53 students, 98%). The vast majority of students (43 students, 79%) noted that in the future it is necessary to organize such classes more often. Conclusions Student responses indicate that group method classes can be used cautiously to study general e-bryology along with traditional didactic lectures., Заохочуючи участь студентів у навчальному процесі, автори внесли зміни до методу групового навчання, згідно з яким змінили викладення по двом темам із загальної ембріології. Метою дослідження була оцінка сприйняття студентами даного методу у порівнянні з лекційною формою навчання. Для цього на кафедрі анатомії медичного коледжу Індії було використано метод анкетування. Матеріали та методи. В цілому у загальному курсі ембріології було виділено вісім розділів. Вивчення шести з них проводилось з використанням традиційного дидактичного методу лекцій. Засвоєння двох інших розділів здійснювалось за допомогою модифікованого методу групового навчання. Студентів було розділено на п'ять груп, кожна з яких отримала комплект матеріалів, що містив базові фактичні відомості, чотири клінічні історії хвороби та попередні екзаменаційні питання по темам. Під час обговорення, що відбувалося в присутності викладача, студенти мали змогу оцінити заняття за допомогою анкет. Результати. Так, більшість студентів відзначили, що навчання за допомогою модифікованого групового методу полегшило виконання завдань (46 студентів, 85%), дозволило краще розуміти учбовий матеріал (43 студента, 79%), стало більш цікавими (43 студента, 81%), забезпечило масову участь студентів у навчальному процесі (51 студент, 94%) та докладання з їх боку більших зусиль (53 студента, 98%). Переважна більшість студентів (43 студента, 79%) зазначили, що в майбутньому необхідно частіше організовувати такі заняття. Висновки. Відповіді студентів вказують на те, що заняття за груповим методом можуть бути вправно використані для вивчення загальної ембріології поряд з традиційними дидактичними лекціями.

Background: The ligaments in coherence with the capsule of the hip joint are known to contribute to hip stability. Nevertheless, the contribution of the mechanical properties of the ligaments and gender- or side-specific differences are still not completely clear. To date, comparisons of the hip capsule ligaments to other tissues stabilizing the pelvis and hip joint, e.g. the iliotibial tract, were not performed. Materials & Methods: Hip capsule ligaments were obtained from 17 human cadavers (9 females, 7 males, 13 left and 8 right sides, mean age 83.65 ± 10.54 years). 18 iliofemoral, 9 ischiofemoral and 17 pubofemoral ligaments were prepared. Uniaxial stress-strain properties were obtained from the load-deformation curves before the secant elastic modulus was computed. Strain, elastic modulus and cross sections were compared. Results: Strain and elastic modulus revealed no significant differences between the iliofemoral (strain 129.8 ± 11.1%, elastic modulus 48.8 ± 21.4 N/mm2), ischiofemoral (strain 128.7 ± 13.7%, elastic modulus 37.5 ± 20.4 N/mm2) and pubofemoral (strain 133.2 ± 23.7%, elastic modulus 49.0 ± 32.1 N/mm2) ligaments. The iliofemoral ligament (53.5 ± 15.1 mm2) yielded a significantly higher cross section compared to the ischiofemoral (19.2 ± 13.2 mm2) and pubofemoral (15.2 ± 7.2 mm2) ligament. No significant gender- or side-specific differences were determined. A comparison to the published data on the iliotibial tract revealed lower elasticity and less variation in the ligaments of the hip joint.

Introduction: Though xenogeneic acellular scaffolds are frequently used for surgical reconstruction, knowledge of their mechanical properties is lacking. This study compared the mechanical, histological and ultrastructural properties of various native and acellular specimens. Materials and methods: Porcine esophagi, ureters and skin were tested mechanically in a native or acellular condition, focusing on the elastic modulus, ultimate tensile stress and maximum strain. The testing protocol for soft tissues was standardized, including the adaption of the tissue’s water content and partial plastination to minimize material slippage as well as templates for normed sample dimensions and precise cross-section measurements. The native and acellular tissues were compared at the microscopic and ultrastructural level with a focus on type I collagens. Results: Increased elastic modulus and ultimate tensile stress values were quantified in acellular esophagi and ureters compared to the native condition. In contrast, these values were strongly decreased in the skin after acellularization. Acellularization-related decreases in maximum strain were found in all tissues. Type I collagens were well-preserved in these samples; however, clotting and a loss of cross-linking type I collagens was observed ultrastructurally. Elastins and fibronectins were preserved in the esophagi and ureters. A loss of the epidermal layer and decreased fibronectin content was present in the skin. Discussion: Acellularization induces changes in the tensile properties of soft tissues. Some of these changes appear to be organ specific. Loss of cross-linking type I collagen may indicate increased mechanical strength due to decreasing transverse forces acting upon the scaffolds, whereas fibronectin loss may be related to decreased load-bearing capacity. Potentially, the alterations in tissue mechanics are linked to organ function and to the interplay of cells and the extracellular matrix, which is different in

Background: To ensure adequate function after orthopedic tumor reconstruction, it is important to reattach the remaining soft tissue to the implant. This study aimed at obtaining mechanical properties of textile muscle-implant and muscle-bone connections in a preliminary test. Methods: Two groups of soft-tissue attachment were mechanically tested and compared: Native bone-muscle samples obtained from human femora and muscles attached to a prosthetic implant by means of Trevira® attachment tubes. Additionally, muscle samples were tested with muscle fibers aligned parallel and perpendicular to the tension load. A uniaxial load was exerted upon all samples. Results: Failure loads of 26.7 ± 8.8 N were observed for the native bone-muscle group and of 18.1 ± 9.9 N for the Trevira® group. Elongations of 94.8 ± 36.2 % were observed for the native bone-muscle group and 79.3 ± 51.8 % for the Trevira® group. The location of failure was mainly observed in the central area of the muscle fibers. Muscle fibers with parallel fiber orientation (47.6 ± 11.5 N) yielded higher tensile strength than those with perpendicular fiber orientation (14.8 ± 4.1 N). Conclusions: Our experiments showed that higher forces were transmitted in the origin and insertion areas than in areas of flat soft tissue reconstruction using attachment tubes. The data indicate that the tested material allows reattaching muscles, but without reinforcing the insertion site. Therefore, attachment tubes with region-dependent and potentially anisotropic material behavior might be advantageous to optimize muscle-bone load transmission after surgery, which may allow lower complication rates and shorter physical recovery.

This study investigated whether mixture extract of Pterocarpus indicus, Momordica charantia, Phaseolus vulgaris and Andrographis paniculata could lower plasma glucose in Type 2 Diabetes Mellitus (T2DM) patients. 43 T2DM patients, consist of 10 male and 33 female patients participated in this possitive-controlled, double-blind, and crossover clinical study with administration of either mixture extract (22 mg/kg BW) or glibenclamide 5 mg daily in breakfast time. Treatment of extract or glibenclamide was administered for a month, then medication was changed after a week of washout period and finally combination therapy was administered after wash out period as well. The efficacy of mixture extracts was measured by using Fasting Plasma Glucose (FPG) and two-hour Postprandial Plasma Glucose (PPG). Forty-one subjects completed the study. Data was analyzed using SPSS 19 with Student's t test, and p<0.05 was considered significant. FPG level significantly decreased 16.07 mg/dl after extract, 43.34 mg/dl after glibenclamide and 50.16 mg/dl after combination treatment. Two-hour PPG level decreased 25.88 mg/dl after extract, 66.61 after glibenclamide and 58.93 after combination treatment. We concluded that extract administration could lower FPG and PPG although not as good as glibenclamide treatment did and combination treatment was the best to lower FPG.

In the last decade the advent of ART has proven to be a boon for the infertile couple. In a large number of infertile couples there may be a genetic basis. Such couples who harbor genetic abnormalities need to be provided comprehensive counseling prior to opting for ART. Despite state of art technology and professional expertise, the carry home live-birth rate in ART is low. One of the factors responsible for low success rate of ART in couples opting for ART could be genetic. We analyzed 350 couples with recurrent spontaneous abortions (RSA) among whom 86 were couples with recurrent ART/ICSI failure. All couples with 3 or more ART failures were referred for cytogenetic, Yq microdeletion and mitochondrial mutation analysis. Among these 86 couples cytogenetic abnormalities were detected in 32 (20 cases in the male and 12 cases in the female). Yq microdeletions were detected from genomic DNA isolated from blood in 7 men whereas 9 men showed Yq microdeletions in DNA isolated from spermatozoa. High frequency of mitochondrial mutations and deletion of 4977 were also detected in sperm DNA (n=3) and blood of the female partner (n=4). These genetic abnormalities may be iatrogenically transmitted to the offspring. In cases with sex chromosomal and autosomal aberrations there is probability of poor embryo development and, consequently, implantation failure and early fetal loss. ART is a very expensive technique and recurrent ART/IVF failure would result in severe financial burden and physical stress, coupled with emotional stress. It is suggested that all couples opting for ART must undergo genetic analysis.

To investigate the association of calmodulin (CaM) with microtubules (MTs) in the mitotic apparatus (MA), the distributions of CaM and tubulin were examined in cells in which the normal spindle organization had been altered. A fluorescent CaM conjugate with tetramethylrhodamine isothiocyanate (CaM-TRITC) and a dichlorotriazinyl aminofluorescein conjugate with tubulin (tubulin-DTAF) were injected into cells that had been treated with the MT inhibitor nocodazole. With moderate nocodazole concentration (0.3 ??g/ml, 37??C, 4 h) in live cells, CaM-TRITC and tubulin-DTAF concentrated identically on or near the centrosomes and kinetochores. In serial sections of these cells, small MT segments were observed by transmission electron microscopy (TEM) in the regions where fluorescent protein had concentrated. When a higher drug concentration was used (3.0 ??g/ml, 37??C, 4 h), no regions of CaM-TRITC or tubulin-DTAF localization were observed, and no MTs were observed when serial sections were examined by TEM. However, following release from the high-concentration nocodazole block, CaM-TRITC colocalized with newly formed MTs at the kinetochores and centrosomes. Later in the recovery period, when chromosome-to-pole fibers had formed, CaM association with kinetochores diminished, ultimately attaining its normal pole-proximal association with kinetochore MTs in cells that progressed through mitosis. We interpret these observations as supporting the hypothesis that in the MA, CaM attains a physical association with kinetochore MTs and suggest that CaM-associated MTs may be inherently more stable.

Samples of human plantar and palmar skin were excised and incubated in 20 mM EDTA after which the epidermis was gently separated from the dermis with the plane of separation occurring in the lamina lucida. Scanning electron microscopic examination of the dermal component revealed the classically described series of regularly spaced grooves and papillae that characterize the epidermal-dermal junction in thick skin. Primary dermal grooves exhibited evenly spaced tunnels that were originally occupied by sweat gland ducts. The basement membrane (basal lamina) in the primary grooves was relatively smooth but did exhibit a flattened, reticulated pattern at high magnifications. The basement membrane of secondary dermal grooves and papillae was in the form of numerous, elevated microridges off of which septae arose at roughly right angles. The surface appearance of the basement membrane in these areas was that of a honeycomb owing to the numerous compartments and recesses formed by the ridges and septae. Degradation of the basement membrane by trypsin demonstrated that the foundation for the highly folded and compartmentalized basement membrane was composed of dermal collagen fibrils, 60–70 nm in diameter, that were arranged in a series of variably sized, interconnected collagen bundles or walls. Epidermal basal cells extended cytoplasmic (foot) processes into two or more compartments, formed by the ridges and septae, which considerably amplified the basement membrane surface available for epidermal attachment. Scanning electron microscopic studies of the epidermal-dermal junction confirm the variable surface character of this interface previously reported by others using sectioned material. This regional variation in surface architecture apparently distinguishes between areas in which epidermal basal cells are specialized for attachment (papillae, secondary dermal grooves) and regions occupied by slow cycling epidermal stem cells from which mitotically active keratinocytes

The objective of this study was to determine how long myogenic cells can survive in the central ischemic zone of early free muscle grafts in the rat. The study was conducted on free grafts of a large (rectus femoris) and a small (extensor digitorum lon-gus) muscle. At times ranging from zero hr to five days post-grafting, the central zones were isolated, minced, and implanted under the back skin of mice. After five days the minces were removed and examined histologically for the presence of rat myotubes, which should form only in minces that contain viable myogenic cells. The results show that myogenic cells survive two to four hr in the ischemic centers of the large rectus femoris grafts; after longer post-grafting intervals, rat myotubes did not arise in central zone minces. In grafts of small muscles, myotubes consistently appeared in central zone minces. Since the formerly ischemic central areas of rectus muscle grafts are ultimately replaced by regenerating muscle fibers, we conclude that these regenerating muscle fibers are derived from precursor cells located outside of the ischemic zone.

Patterns of activity of six dehydrogenase enzymes were studied histochemically in 42 free muscle grafts in the rat. Within hours, the surviving peripheral muscle fibers can be distinguished from the central ischemic muscle fibers. The surviving muscle fibers retain their characteristic pattern of staining throughout the post-transplantation period. The central ischemic muscle fibers stain abnormally and by five or six days they lose their enzymatic activity. The zone of regeneration, between the surviving and the ischemic muscle fibers, initially shows little dehydrogenase activity, but as the regenerating muscle fibers mature, they develop first a homogeneous staining pattern and, later, differences in staining intensity among different types of muscle fibers.

Since differential chemical staining has been unsatisfactory for demonstration of specific secretory cell types in the hypophyseal pars distalis of the mouse, the objective of this study was to determine whether the peroxidase-antiperoxidase immunocytochemical procedure might be more effective. Accordingly, representative sections from the hypophyses of 17 female and 15 male adult mice of the Swiss-Webster strain were immunostained, 16 antisera to 5 pituitary hormones or their subunits being utilized. Five secretory cell types were demonstrated. Somatotropes were ovoid to spheroidal and distributed quite generally in the gland except for the ???sex zone??? where they were scarce. Somatotropes were larger and more numerous in the male than in the female. Mammotropes were polyhedral and also generally distributed in the gland except for the ???sex zone??? where few were observed. Mammotropes were larger and more numerous in the female than in the male. Corticotropes were small, stellate and few. They were most common near the ventral surface of the gland and formed bilateral centromedial groups in the lateral wings. Thyrotropes were usually large and polyhedral. They were restricted almost solely to the ventral region of the pars distalis. Gonadotropes were polyhedral, and generally distributed, except for aggregation in the cephalomedian ???sex zone.??? Most gonadotropes appeared to contain both luteinizing hormone and follicle-stimulating hormone. Thus, all secretory cell types recognized in other species can be demonstrated readily in the mouse hypophysis with immunocytochemistry.

Cone photoreceptors in the zebrafish retina are arranged in a crystalline lattice, with each spectral subtype at a specific position in the array: rod photoreceptors are inserted around the cones. Patterning events and developmental mechanisms that lead to the formation of the cone mosaic are not known. To begin investigating this issue, we examined the initial stages of opsin expression in zebrafish embryos by in situ hybridization with goldfish opsin cRNA probes to determine how and when the cone mosaic pattern arises. We found both differences and similarities in the spatiotemporal patterns of rod and cone development, which suggest the following: (1) Expression of opsin message (including rod opsin, blue and red cone opsins) was initiated at 50–52 hours postfertilization by a few photoreceptors which were consistently found in a ventral patch of retina located nasal to the choroid fissure. (2) The cone mosaic pattern was generated by a crystallization-like process initiated in the precocial ventral patch and secondarily in nasal retina, which then swept like a wave into dorsotemporal retina. (3) The pattern of differentiation of rods in the ventronasal patch differed substantially from that in the remainder of the retina, suggesting that these precocial rods might differ from typical rods. (4) Developmental maturation of rods in zebrafish, as reflected by expression of opsin, may be accelerated compared to cones, which are thought to become postmitotic before rods. These data are consistent with a model in which lateral inductive interactions among differentiating photoreceptors lead to patterning of the array. © 1995 Wiley-Liss, Inc.

The process of revascularization of free grafts of the extensor digitorum longus muscle in rats has been studied by gross, histological and electron microscopic methods. During the first day after transplantation the muscle is entirely avascular, and it consists of a thin peripheral zone of surviving muscle fibers and a large central area of ischemic muscle. The original blood vessels of the graft undergo a sequence of intrinsic and cell-mediated destruction. Scattered sinusoidal vessels begin to grow into the graft starting on the second day, and ingrowing blood vessels progressively invade the deeper tissues of the graft. Most new vessels form in the connective tissues, but some vessels, especially larger ones, grow into persisting basal laminae from preexisting vessels and, occasionally, even into basal laminae from degenerated muscle fibers and nerves. The differentiation of new arterioles and venules in free muscle grafts is described. By the end of the first week, the entire graft is revascularized, and ultimately a fairly normal relationship between new capillaries and regenerating muscle fibers is established. In mature grafts, however, irregularities are sometimes found in the organization of smooth muscle cells associated with larger vessels.

Our radioimmunoassay data reveal that ovariectomy for either one or two months resulted in a significant decrease in the GnRH content of the mouse basal hypothalamus, while in mice ovariectomized for two weeks the GnRH levels did not differ significantly from controls. Parallel immunocytochemical analyses gave similar results. Tissue sections from specific regions of the median eminence in intact and ovariectomized mice were immunolabeled for GnRH with the peroxidase-antiperoxidase procedure of Sternberger. Differences in area and average unit density of the reaction product were measured with a Quantiment 720 image analyzer. At two weeks after ovariectomy, a majority of matched sections from intact and ovariectomized animals, taken in each sampled region of the median eminence, showed no visually apparent difference in GnRH immunoreactivity. Quantitative image analysis of these preparations revealed no statistically significant difference in either the area of neural tissue covered by deposit or the average unit density of this material in any given region. In contrast, in mice ovariectomized for either one or two months, immunolabeled GnRH was decreased throughout the cephalo-caudal extent of the median eminence. In each region, a reduction in reaction deposit was observed over most of the area in which GnRH is localized, especially in the medial aspects of this distribution. Image analysis of these preparations revealed a significant reduction of both specimen area immunolabeled for GnRH and the average unit density of reaction product in each region of the median eminence. While the decrease in density was uniform throughout the median eminence, the greatest reduction in area covered by deposit occurred in the infundibular region. The demonstration of a decrease in basal hypothalamic GnRH in the chronically ovariectomized mouse suggests that GnRH neurosecretion is increased in response to the absence of the ovaries and supports the premise that tonic secretion o

Values are presented for body constants based on a study of nine male white cadavers of normal appearance and average build. The limb data are supplemented by a further analysis of 11 upper and 41 lower limbs. Techniques used in the study form standard procedures that can be duplicated by subsequent workers. Each cadaver was measured, weighed, and somatotyped. Joints were placed in the midposition of the movement range and the body was frozen rigid. Joint angles were bisected in a systematic dismemberment procedure to produce unit segments. These segment lengths were weighed, measured for linear link dimensions, and analysed for segment volumes. The segment centers of mass were located relative to link end points as well as in relation to anatomical landmarks. Finally, each segment was dissected into its component parts and these were weighed. The specific gravity of each body part was calculated separately. Data are expressed in mean values together with standard deviations and, where available, are correlated and evaluated with other values in the literature.

Amphibian cardiac myocytes are predominantly mononucleated and have been demonstrated to respond to injury with DNA synthesis and mitosis. The nature of this response with regard to nuclear number and ploidy is unclear. In this study, the apex of the newt ventricle was minced and replaced, increasing the reactive area of the wound. At 45 days after mincing following multiple injections of tritiated thymidine (2.5-??Ci/animal, 20 Ci/mM) 15 to 20 days after mincing, three ventricular zones were isolated and fixed: Zone 1, the minced area; Zone 2, extending approximately 500??m proximally from the amputation plane; and Zone 3, the portion proximal to Zone 2. Myocytes separated in 50% KOH were examined for DNA synthesis by autoradiography and for nuclear number and DNA content using a scanning microdensitometer on Feulgen-Naphthol yellow S-stained cells. No labeled myocyte nuclei were found in control hearts and 98.3% of the myocytes were 2C. At 45 days, 46.78% of myocyte nuclei within Zone 1 were labeled, while 13% were non-diploid. In Zone 2, 9.25% were labeled with 4.8% non-diploid. In Zone 3, 1.1% were labeled, with 2.8% non-diploid. The newt ventricle's response to injury apparently may involve complete mitosis and cytokinesis, resulting in mononucleated diploid cells.

In this study, we have compared spatial and temporal aspects of development of new rods and cones in the adult goldfish by using a combination of bromodeoxyuridine immunocytochemistry and opsin in situ hybridization to determine the intervals between terminal mitosis (cell "birth") and expression of opsin mRNA for each photoreceptor cell type. The goldfish opsins include rod opsin and four different cone opsins: red, green, blue, and ultraviolet. In a cohort of photoreceptors born at the same time, rods expressed opsin mRNA within 3 days of cell birth, while expression of cone opsin mRNA required at least 7 days. This temporal discrepancy in differentiation, coupled with a discordance in the site of cell genesis of rods and cones, allowed opsin expression to commence in both cell types in approximately the same retinal location. Commitment to the generic cone phenotype occurred within approximately 6 days throughout the cone cohort, as indicated by expression of interphotoreceptor retinoid-binding protein (IRBP) mRNA, but expression of a specific spectral phenotype was delayed until rods differentiated nearby. Onset of expression of cone opsin mRNA followed a phenotype-specific sequence: red, then green, then blue, and finally ultraviolet; in situ hybridization with two opsin probes confirmed that individual photoreceptors expressed only one type of opsin as they differentiated. This stepwise process of cone differentiation is consistent with the hypothesis that cell-cell interactions among developing photoreceptors may coordinate selection of specific photoreceptor phenotypes. J. Comp. Neurol. 382:272-284, 1997. © 1997 Wiley-Liss, Inc.

Cell adhesion molecules in the cadherin family have been implicated in histogenesis and maintenance of cellular structure and function in several organs. Zebrafish have emerged as an important new developmental model, but only three zebrafish cadherin molecules have been identified to date (N-cadherin, paraxial protocadherin, and VN-cadherin). We began a systematic study to identify other zebrafish cadherins by screening zebrafish cDNA libraries using an antibody raised to the cytoplasmic domain of mouse E-cadherin. Here, we report a partial cDNA with extensive sequence homology to R-cadherin. Spatial and temporal expression of this putative zebrafish R-cadherin was examined in embryos and adults by Northern analysis, RNase protection, and in situ hybridization. R-cadherin message increased during embryogenesis up to 80 hours postfertilization (hpf) and persisted in adults. In the embryonic brain, R-cadherin was first expressed in groups of cells in the diencephalon and pretectum. In adult zebrafish brain, R-cadherin continued to be expressed in several specific regions including primary visual targets. In the retina, R-cadherin was first detected at about 33 hours postfertilization in the retinal ganglion cell layer and the inner part of the inner nuclear layer. Expression levels were highest during periods of axon outgrowth and synaptogenesis. Retrograde labeling of the optic nerve with 1,12 dioctadecyl-3,3,32 32 tetramethylindocarbocyanine perchlorate (DiI) followed by in situ hybridization confirmed that a subset of retinal ganglion cells in the embryo expressed R-cadherin message. In the adult, R-cadherin expression continued in a subpopulation of retinal ganglion cells. These results suggest that R-cadherin-mediated adhesion plays a role in development and maintenance of neuronal connections in zebrafish visual system. J. Comp. Neurol. 410:303-319, 1999. © 1999 Wiley-Liss, Inc.

The primary purpose of the present study was to determine whether a rhodopsin-like gene, which has been postulated to represent the green cone pigment in several species, is in fact expressed in cone photoreceptors instead of rods. The expression patterns of rod opsin and blue and red cone opsins were also examined in both goldfish and zebrafish retinas using colorimetric in situ hybridization. The results demonstrate that the rhodopsin-like gene is expressed in green cones, as predicted. A subset of small cones that do not hybridize with these cRNA probes are tentatively identified as ultraviolet receptors. The results also demonstrate that opsin message in cones is restricted to the perinuclear region, whereas in rods, it is both perinuclear and adjacent to the ellipsoid.

The influence of chronic consumption of fresh and thermoxidized plam oil diets on sperm count and motility was studied in rats. Thirty young Wister rats initially weighing between 50-65g were randomly divided into 3 group, (A,B and C) of ten rats each. Group A rats were fed thermoxidized palm oil diet, obtained by mixing 15g of thermoxidized palm oil with 85g of rat feed (15% w/w). Group B rats were fed fresh palm oil diet, obtained by mixing 15g of fresh palm oil with 85g of rat feed (15% w/w). Group C rats served as control and were allowed fed normal rat fed. The animals were allowed the test diets and water ad libitum for 20 weeks. Results obtained showed that there was a significant difference (p

Vitexin, identified as apigenin-8-C-D-glucopyranoside, a natural flavonoid compound found in certain herbs such as hawthorn herb. The aim of this study is to investigate the regulation of glycolysis underlying vitexin-induced anti-adipogenesis in 3T3-L1 adipocytes. Vitexin inhibited adipose accumulation, glucose consumption and triglyceride synthesis. The expression of ERK 1/2 MAPK greatly induced, whereas the expression of adipogenic markers Akt and peroxisome proliferator-activated receptor γ (PPARγ) diminished. ERK 1/2 MAPK inhibitor PD98059 (10 μM) significantly enhanced lipid accumulation, triglyceride synthesis and the expression of adipogenic markers. However, PD98059 had no effect on glucose consumption. In regulation of glycolysis, vitexin induced the expression of glycerol-3-phosphate dehydrogenase (GPDH) at higher dosage (50 and 100 μM) and without any effect on glucose-6-phosphate dehydrogenase (G6PDH) expression. PD98059 had an opposing effect that it significantly increased the expression of G6PDH, but decreased the expression of GPDH. Vitexin increased the lactic acid synthesis to about 3.8 folds in concentration dependent, whereas, PD98059 decreased the level of lactic acid in media to about 72% when compared with controls. Moreover, PD98059 abolished the anti-adipogenic effect of vitexin. Vitexin influence the expansion of adipose tissue through its ability to inhibit preadipocyte migration to about 80% via decreased the activity of active MMP-2. We demonstrated evidences for the first time that using of vitexin to against adipose accumulation, at least in part, was regulation of glycolysis via activation of ERK 1/2 MAPK signaling. Moreover, blockade pentose phosphate pathway may be a novel strategy for obesity prevention and therapy.

The aim of the study was to investigate the effects of Ficus platyphylla on female Wistar rats with estradiol valerate (EV)-induced polycystic ovarian syndrome (PCOS). PCOS was induced in female Wistar rats with regular 4-5 day oestrus cycles via the intramuscular injection of 4 mg EV per rat in an oily solution. The rats were allowed 30 days to establish PCOS. Then, the animals were divided into five treatment groups. The positive control group received clomiphene citrate, the negative control group received distilled water and the other groups received 100, 200 and 400 mg/kg of the aqueous extract. All groups were dosed for 15 days, except the positive control group, which was dosed for 5 days. On the 16th day, the animals were sacrificed. Hormonal assays and histological studies were then conducted. An elevated Luteinizing hormone (LH)-to- Follicle stimulating hormone (FSH) ratio is typical in many women with PCOS. The extract-treated groups showed a lower LH/FSH ratio compared with both the positive and the negative control groups. The progesterone levels were higher in the extract-treated groups compared with the negative control group, indicating luteal phase repair. The influence of the extract on ovarian morphology in the EV-induced PCOS model showed a marked reversal of polycystic ovary. These data help validate the use of this extract in folk medicine for the treatment of infertility. Considering its apparent safety, this study paves the way for an efficacy study for its use in treatment of PCOS in humans.

The cytotoxic effects of Teucrium polium (Lamaceae) have been shown in some studies. However, the antitumor activity of the fractions of this herb has not been studied, neither has the pattern of cell death been evaluated yet. The inhibitory effect of Teucrium polium (T. polium) on U87 cells was evaluated by treatment of U87 cells with various concentrations of crude extract (CE), a petroleum ether (PE) fraction and a diethyl ether (DE) fraction of T. polium for 24 and 48 h. Trypan blue and differential staining were also used to assess the cell viability and the pattern of cell death. CE, PE and DE inhibited cell growth in a dose-dependent manner, and this inhibition was accomplished by necrotic cell death, as assessed by differential staining. The highest inhibitory effect (IC50=64.47) was demonstrated by the PE fraction of T. polium followed by the DE fraction and the CE. Cell death was also higher when U87 cells were treated with PE (P<0.001 compared with other treated groups). Necrotic cells were predominant in all treated groups including CE, PE and DE. The results suggest that the PE fraction of T. polium is the most potent against U87 cells. The antitumor effect of the PE fraction of T. polium is probably executed through the necrotic cell death mechanism.

Introduction : For more than a century, the temporalis muscle has been used for facial reconstructions. More recently, a split temporalis muscle flap elevated on the superficial temporal pedicle has been described, for which the resulting gain of length makes crossing of the midline possible, as well as reconstruction of substance losses exceeding the midline. Materials and methods : Fourteen fresh cadaveric dissections were performed to study the different techniques for splitting the temporalis muscle. Dissections with catheterization and injection of radio-opaque contrasting agent in the external carotid artery were then performed to specify the vascularization of the flap split on the superficial temporal pedicle. Results : The duplication of the superficial temporal pedicle grants greater length compared to that of the deep pedicles, 57 mm versus 40 (p = 0.036). The middle temporal artery is capable of ensuring the vascularization, and therefore the viability, of the split flap. From these results, we spoke about the limitations of this study and we have inferred the main indications.

Urtica dioica (Stinging nettle) leaves have been used as a traditional medicine to treatrheumatoid arthritis, to alleviate rheumatic pain benign prostatic hyperplasia (BPH), diuretic and prevention of kidney stones by local people in Middle East region especially in Iran, Turkey and East Europe. Hence, this investigation evaluates the antidepressant effect of selected crude extracts in the forced swimming test (FST) and tail suspension test (TST), two models predictive of antidepressant activity. The acute treatment of mice with extracts by intra-peritoneal (i.p.) route significantly reduced the immobility time in the FST (50 and 100 mg/kg) and TST (50 and 100 mg/kg), as compared to positive controls (haloperidol and fluoxetine) at 1 and 10 mg/kg, respectively. On the third day of experiment, a significant decrease of mobility was observed for chloroform extract (CE I) and butanol extract (BE II) compared to first day. CE I and BE II led to reduction of immobility time, as the selected extracts with two doses administered were different compared to the control, in the FST method by 65.37% and53.92% for 100 mg/kg, respectively. However, CE I showed the best result compared to our positive controls. Similar results of increased antidepressant effect, that was, of immobility time depending on the concentration administered, were obtained with the TST method. Also our data showed that there was no significant differences between doses (50 and 100 mg/kg). The results suggested that the antidepressant action of the butanol extract and of U. dioica its fraction (BE II2) was mediated by an interaction with 5-hydroxytrptamine (5-HT). U. dioica showed a potential source for the isolation of important natural products with antidepressant-like properties. However, further studies are still required

Daptomycin is a lipopeptide antibiotic with strong bactericidal effects against Gram-positive bacteria and minor side effects on skeletal muscles. The type and magnitude of the early effect of daptomycin on skeletal muscles of rats was quantified by histopathology, examination of contractile properties, Evans Blue Dye uptake, and effect on the patch repair process. A single dose of daptomycin of up to 200 mg/kg had no effect on muscle fibers. A dose of 150 mg/kg of daptomycin, twice per day for 3 days, produced a small number of myofibers (???0.22%) with loss of plasma membrane integrity and/or infiltration by neutrophils and/or macrophages. Multiple doses of daptomycin are required for a quantifiable effect on skeletal muscles of rats. Some fibers were Evans Blue Dye???positive but were not yet infiltrated by neutrophils. This suggests that the sarcolemma may be the primary target for the observed effects. Muscle Nerve, 2010

Vascular injection of the macromolecular tracer, horseradish peroxidase (HRP), was used to study the permeability of the odontoblast cell layer in developing and mature rat molar teeth, and to investigate the effect of cavity preparations on the permeability of this epithelioid cell layer in adult animals. HRP injected into the vascular system of normal animals 28 days of age and older was localized histochemically (from 5 to 90 min after injection) throughout the extracellular spaces of the maxillary dental pulps; however, the tracer did not penetrate beyond the tight junctions at the apical region of the odontoblast cell layer, and was absent from the predentin and dentin. In contrast, HRP injected into very young neonatal animals (e.g., day 3) resulted in free passage of HRP between odontoblasts and into the overlying predentin and dentin. When Class V cavities had been prepared in adult maxillary molars after HRP was injected into the blood stream, HRP reaction product penetrated the predentin and dentin immediately beneath the cavity preparation; however, adjacent, un-traumatized areas of predentin and dentin in the operated teeth were devoid of reaction product. These results provide evidence that: (1) a physiological barrier develops between the distal segments of odontoblast cell bodies in normal rat molar teeth between days 15 and 28 of postnatal life, and this barrier prevents the passage of macromolecules from the pulp into the predentin and dentin; and (2) this barrier is perturbed following routine restorative procedures in adult animals.