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Structural alterations in the seminiferous tubules of rats treated with immunosuppressor tacrolimus.Reproductive Biology and Endocrinology... Feb 2009Tacrolimus (FK-506) is an immunosuppressant that binds to a specific immunophilin, resulting in the suppression of the cellular immune response during transplant...
BACKGROUND
Tacrolimus (FK-506) is an immunosuppressant that binds to a specific immunophilin, resulting in the suppression of the cellular immune response during transplant rejection. Except for some alterations in the spermatozoa, testicular morphological alterations have not been described in rats treated with tacrolimus. In the present study, we purpose to evaluate if the treatment with tacrolimus at long term of follow-up interferes in the integrity of the seminiferous tubules.
METHODS
Rats aging 42-day-old received daily subcutaneous injections of 1 mg/kg/day of tacrolimus during 30 (T-30) and 60 (T-60) days; the rats from control groups (C-30 and C-60) received saline solution. The left testes were fixed in 4% formaldehyde and embedded in glycol methacrylate for morphological and morphometric analyses while right testes were fixed in Bouin's liquid and embedded in paraffin for detection of cell death by the TUNEL method. The epithelial and total tubular areas as well as the stages of the seminiferous epithelium and the number of spermatocytes, spermatids and Sertoli cells (SC) per tubule were obtained.
RESULTS
In the treated groups, seminiferous tubules irregularly outlined showed disarranged cellular layers and loss of germ cells probably due to cell death, which was revealed by TUNEL method. In addition to germ cells, structural alterations in the SC and folding of the peritubular tissue were usually observed. The morphometric results revealed significant decrease in the number of SC, spermatocytes, spermatids and significant reduction in the epithelial and total tubular areas.
CONCLUSION
Tacrolimus induces significant histopathological disorders in the seminiferous tubules, resulting in spermatogenic damage and reduction in the number of Sertoli cells. A careful evaluation of the peritubular components will be necessary to clarify if these alterations are related to the effect of FK-506 on the peritubular tissue.
Topics: Animals; Body Weight; Immunosuppressive Agents; In Situ Nick-End Labeling; Male; Organ Size; Rats; Rats, Sprague-Dawley; Seminiferous Tubules; Tacrolimus; Testis
PubMed: 19243597
DOI: 10.1186/1477-7827-7-19 -
The Journal of Veterinary Medical... Mar 2014The extracellular matrix of the cervix that comprises collagen, elastin, proteoglycans and glycosaminoglycans (GAGs) is thought to have an essential role in cervical... (Comparative Study)
Comparative Study
The extracellular matrix of the cervix that comprises collagen, elastin, proteoglycans and glycosaminoglycans (GAGs) is thought to have an essential role in cervical relaxation. This study investigated the proportion of collagen and smooth muscle as well as the GAGs in cervices obtained from healthy bitches at different stages of the estrous cycle and bitches with open- and closed-cervix pyometra. Cervices were collected after ovariohysterectomy. The proportion of collagen to smooth muscle was determined using Masson's trichrome staining. Alcian blue staining was used to evaluate the relative distribution of cervical GAGs. The proportion of cervical collagen relative to smooth muscle was higher at estrus compared to anestrus (P≤0.05). It was also higher (P≤0.05) in bitches with open- compared to those with closed-cervix pyometra. Overall, hyaluronan (HA) was the predominant GAG in the canine cervix. In the luminal epithelium, the staining intensity for HA was stronger in estrus than in anestrus (P≤0.05), but not in diestrus (P>0.05). On the contrary, the intensity for the combined keratan sulfate (KS) and heparan sulfate (HS) was stronger in anestrus than in estrus and diestrus (P≤0.05). In bitches with pyometra, the staining intensity of the stroma for KS and HS was weaker in open- compared to closed-cervix pyometra (P≤0.05). Collectively, the different profiles of collagen and GAG suggest that the metabolism of both collagen and GAGs in the canine cervix is associated with hormonal statuses during the estrous cycle and cervical patency of bitches with pathological uterine conditions, such as pyometra.
Topics: Acetic Acid; Alcian Blue; Animals; Azo Compounds; Cervix Uteri; Collagen; Dog Diseases; Dogs; Eosine Yellowish-(YS); Estrous Cycle; Extracellular Matrix; Female; Formaldehyde; Glycosaminoglycans; Immunoassay; Methyl Green; Muscle, Smooth; Picrates; Progesterone; Pyometra
PubMed: 24152876
DOI: 10.1292/jvms.13-0114 -
Alcoholism, Clinical and Experimental... Jun 2009Magnetic resonance microscopy (MRM), magnetic resonance imaging (MRI) at microscopic levels, provides unprecedented opportunities to aid in defining the full spectrum of...
BACKGROUND
Magnetic resonance microscopy (MRM), magnetic resonance imaging (MRI) at microscopic levels, provides unprecedented opportunities to aid in defining the full spectrum of ethanol's insult to the developing brain. This is the first in a series of reports that, collectively, will provide an MRM-based atlas of developmental stage-dependent structural brain abnormalities in a Fetal Alcohol Spectrum Disorders (FASD) mouse model. The ethanol exposure time and developmental stage examined for this report is gestational day (GD) 8 in mice, when the embryos are at early neurulation stages; stages present in humans early in the fourth week postfertilization.
METHODS
For this study, pregnant C57Bl/6J mice were administered an ethanol dosage of 2.8 g/kg intraperitoneally at 8 days, 0 hour and again at 8 days, 4 hours postfertilization. On GD 17, fetuses that were selected for MRM analyses were immersion fixed in a Bouin's/Prohance solution. Control fetuses from vehicle-treated dams were stage-matched to those that were ethanol-exposed. The fetal mice were scanned ex vivo at 7.0 T and 512 x 512 x 1024 image arrays were acquired using 3-D spin warp encoding. The resulting 29 microm (isotropic) resolution images were processed using ITK-SNAP, a 3-D segmentation/visualization tool. Linear and volume measurements were determined for selected brain, head, and body regions of each specimen. Comparisons were made between control and treated fetuses, with an emphasis on determining (dis)proportionate changes in specific brain regions.
RESULTS
As compared with controls, the crown-rump lengths of stage-matched ethanol-exposed GD 17 fetuses were significantly reduced, as were brain and whole body volumes. Volume reductions were notable in every brain region examined, with the exception of the pituitary and septal region, and were accompanied by increased ventricular volumes. Disproportionate regional brain volume reductions were most marked on the right side and were significant for the olfactory bulb, hippocampus, and cerebellum; the latter being the most severely affected. Additionally, the septal region and the pituitary were disproportionately large. Linear measures were consistent with those of volume. Other dysmorphologic features noted in the MR scans were choanal stenosis and optic nerve coloboma.
CONCLUSIONS
This study demonstrates that exposure to ethanol occurring in mice at stages corresponding to the human fourth week postfertilization results in structural brain abnormalities that are readily identifiable at fetal stages of development. In addition to illustrating the utility of MR microscopy for analysis of an FASD mouse model, this work provides new information that confirms and extends human clinical observations. It also provides a framework for comparison of structural brain abnormalities resulting from ethanol exposure at other developmental stages and dosages.
Topics: Abnormalities, Drug-Induced; Animals; Brain; Central Nervous System Depressants; Cerebellum; Disease Models, Animal; Dose-Response Relationship, Drug; Embryonic Development; Ethanol; Female; Fetal Alcohol Spectrum Disorders; Fetus; Hippocampus; Magnetic Resonance Imaging; Mice; Mice, Inbred C57BL; Olfactory Bulb; Pituitary Gland; Pregnancy
PubMed: 19302087
DOI: 10.1111/j.1530-0277.2009.00921.x -
Poultry Science Mar 2007The objective of this experiment was to evaluate the influence of Gln and vitamin E (VE) supplementation in the diet of broiler chickens (Cobb-Vantress) on the... (Randomized Controlled Trial)
Randomized Controlled Trial
The objective of this experiment was to evaluate the influence of Gln and vitamin E (VE) supplementation in the diet of broiler chickens (Cobb-Vantress) on the morphometry of the intestinal mucosa. The design was completely randomized in a 2 x 3 (VE x periods of administering Gln) factorial arrangement. The levels of VE used were 10 and 500 mg/kg of diet and 3 periods of administering (1%) Gln-supplemented starter diet (for the first 7 or 14 d of life or for no added Gln), totaling 6 treatments with 5 replicates of 50 birds per experimental unit. In the growth period (d 22 to 41 posthatch), the treatments consisted only in the respective levels of VE. On d 7, 14, 21, and 41 posthatch, 2 birds per replicate were killed, and samples of the duodenum, jejunum, and ileum were subsequently removed, fixed in Bouin solution, and later embedded in paraffin and stained with hematoxylin-eosin. The parameters analyzed were villus height and crypt depth. An ANOVA was applied to the obtained data, and the means were compared using Tukey's test (5% significance level). Greater development was observed in the duodenum, followed by the jejunum and ileum. On 41 d of life, diets with 10 mg of VE/kg supplemented with Gln (for the first 7 d of life) provided better development of the intestinal mucosa in broiler chickens.
Topics: Aging; Animal Feed; Animal Nutritional Physiological Phenomena; Animals; Chickens; Diet; Dietary Supplements; Duodenum; Glutamine; Ileum; Intestinal Mucosa; Jejunum; Male; Vitamin E
PubMed: 17297160
DOI: 10.1093/ps/86.3.488 -
Canadian Journal of Gastroenterology =... Nov 2013Many gastroenterology (GI) trainees face a variety of barriers to stable employment and are finding it increasingly difficult to secure employment in their chosen field.
BACKGROUND
Many gastroenterology (GI) trainees face a variety of barriers to stable employment and are finding it increasingly difficult to secure employment in their chosen field.
OBJECTIVE
To elucidate factors that contribute to the burden of unemployment and underemployment, and to examine solutions that may remedy this growing problem in the field of GI.
METHODS
A nationwide survey of current, incoming and recently graduated individuals of GI training programs in Canada was conducted. Trainees in pediatric GI programs and those enrolled in subspecialty programs within GI were also included.
RESULTS
The response rate was 62%, with 93% of respondents enrolled in an adult GI training program. Many (73%) respondents planned to pursue further subspecialty training and the majority (53%) reported concerns regarding job security after graduation as contributory factors. Only 35% of respondents were confident that they would secure employment within six months of completing their training. Regarding barriers to employment, the most cited perceived reasons were lack of funding (both from hospitals and provincial governments) and senior physicians who continue to practice beyond retirement years. Sixty-nine per cent perceived a greater need for career guidance and 49% believed there were too many GI trainees relative to the current job market in their area. Most residents had a contingency plan if they remained unemployed >18 months, which often included moving to another province or to the United States.
CONCLUSION
GI trainees throughout Canada reported substantial concerns about securing employment, citing national retirement trends and lack of funding as primary barriers to employment. Although these issues are not easily modifiable, certain problems should be targeted including optimizing training quotas, tailoring career guidance to the needs of the population, and emphasizing credentialing and quality control in endoscopy.
Topics: Adult; Canada; Career Choice; Data Collection; Employment; Female; Gastroenterology; Humans; Internship and Residency; Male; Physicians; Specialization; Unemployment
PubMed: 24199210
DOI: 10.1155/2013/950649 -
Brain and Behavior Apr 2023The morphology of senile plaques depends on the APP knock-in mice brain fixative. Solid forms of senile plaques were detected in APP knock-in mice after formic acid...
The morphology of senile plaques depends on the APP knock-in mice brain fixative. Solid forms of senile plaques were detected in APP knock-in mice after formic acid treatment with Davidson's and Bouin's fluid fixative as the brain of AD patients. Aβ42 was deposited as cored plaques and Aβ38 accumulated around Aβ42.
Topics: Mice; Animals; Fixatives; Plaque, Amyloid; Formaldehyde; Brain; Alzheimer Disease; Mice, Transgenic
PubMed: 36879381
DOI: 10.1002/brb3.2953 -
Veterinary Medicine International 2021Mesenchymal stem cells-conditioned medium (MSC-CM) is the extraction from stem cell medium containing biological substances, including growth factors and cytokines....
Mesenchymal stem cells-conditioned medium (MSC-CM) is the extraction from stem cell medium containing biological substances, including growth factors and cytokines. These substances play roles in the various functions of body regulatory, including bone formation. However, the effect of MSC-CM derived from human umbilical cord injection in femur fracture healing of rats has not been reported previously. This study aims to see the effect of MSC-CM derived from human umbilical cord injection on the callus formation of bone fracture healing in Wistar rats (). A femur fracture in 54 Wistar rats was made by surgery according to the procedure under sterile conditions. After the surgery, rats were divided into 2 groups of 27, respectively. Injection in the control (0.1 mL/kg body weight NaCl) and MSC-CM group (0.1 mL/kg body weight MSC-CM) was performed on weeks 0, 1, 2, 3, 4, 5, 6, 7, and 8 after surgery. Radiographic images and the femur bone samples were taken and collected on days 1, 7, 14, 21, 28, 35, and 60 after surgery. Bone samples were then fixed in Bouin solution. Histologic preparations were done by the paraffin method, by cutting the tissue blocks into 5 m thickness and then staining with Mallory aniline blue staining. The results were analyzed descriptively and quantitatively. The result showed that the soft callus formation occurred rapidly and got wider in the MSC-CM group than that of the control group. The administration of MSC-CM injection postfracture surgery to femur fracture cases in rats was capable to accelerate the callus formation.
PubMed: 33996023
DOI: 10.1155/2021/8410175 -
Anais Da Academia Brasileira de Ciencias 2018Chronic stress exposure commonly increases adrenals weight and changes their morphology. This study aimed to compare four methods to delimitate the cortical and...
Chronic stress exposure commonly increases adrenals weight and changes their morphology. This study aimed to compare four methods to delimitate the cortical and medullary layers of adrenals glands in Nelore bulls. Fresh adrenals did not present differentiation between layers. Then, frozen adrenals were distributed in plastics bags with fixative Bouin (G1), 96ºGL ethylic alcohol (G2), 10% formaldehyde (G3), or 2.5% glutaraldehyde (G4). After 12 hours of fixation, the G1 adrenal glands did not show the entire cortical layer marked by Bouin's solution. For G2 and G3 there was a poor contrast, while for G4 there was a reasonable contrast. After 24 hours of fixation, G1 had an excellent contrast between layers, while G2 and G4 had a reasonable contrast and G3 a very bad contrast. After 48 hours it was difficult to differentiate cortical and medullar layers for G1; for Group 2 we get a reasonable contrast; and for G3 the contrast was bad. For G4 the contrast was not as sharp due to the medulla became dark. It was concluded that fixation of adrenals must be done in Bouin's solution for 24 hours to obtain an effective evaluation of the adrenals' morphometry.
Topics: Acetic Acid; Adrenal Glands; Animals; Cattle; Ethanol; Fixatives; Formaldehyde; Glutaral; Picrates
PubMed: 30517224
DOI: 10.1590/0001-3765201820170941 -
Reproductive Toxicology (Elmsford, N.Y.) Apr 2009Toluene is a commonly abused organic solvent. Inhalant abusers are increasingly women in their prime childbearing years. Children born to mothers who abused solvents...
Toluene is a commonly abused organic solvent. Inhalant abusers are increasingly women in their prime childbearing years. Children born to mothers who abused solvents during pregnancy may exhibit characteristics of a "fetal solvent syndrome" which may include dysmorphic features. This study examined the teratological effects of an abuse pattern of binge toluene exposure during gestation on skeletal and soft tissue abnormalities, body weight, and body size in fetal rats. Pregnant Sprague-Dawley rats were exposed for 30 min, twice daily, from gestational day (GD) 8 through GD20 to either air (0 ppm), 8000 ppm, 12,000 ppm, or 16,000 ppm toluene. Two-thirds of each litter was prepared for skeletal examination using Alizarin Red S staining while the remaining third of each litter was fixed in Bouin's solution for Wilson's soft tissue evaluation. Exposure to toluene at all levels significantly reduced growth, including decreases in placental weight, fetal weight, and crown-rump length. In addition, numerous gross morphological anomalies were observed such as short or missing digits and missing limbs. Skeletal examination revealed that ossification of the extremities was significantly reduced as a result of toluene exposure at all levels. Specific skeletal defects included misshapen scapula, missing and supernumerary vertebrae and ribs, and fused digits. Soft tissue anomalies were also observed at all toluene levels and there was a dose-dependent increase in the number of anomalies which included cryptorchidism, displaced abdominal organs, gastromegaly, distended/hypoplastic bladder, and delayed cardiac development, among others. These results indicate that animals exposed prenatally to levels and patterns of toluene typical of inhalant abuse are at increased risk for skeletal and soft tissue abnormalities.
Topics: Abnormalities, Drug-Induced; Animals; Bone and Bones; Crown-Rump Length; Dose-Response Relationship, Drug; Female; Fetal Weight; Fetus; Gestational Age; Inhalation Exposure; Maternal-Fetal Exchange; Models, Animal; Placenta; Pregnancy; Rats; Rats, Sprague-Dawley; Solvents; Substance-Related Disorders; Toluene
PubMed: 19429395
DOI: 10.1016/j.reprotox.2009.01.002 -
The American Journal of Pathology Sep 1989Monoclonal antibodies (mAB) against progesterone receptor (PR) and the peroxidase-antiperoxidase (PAP) method to visualize PR in paraffin sections from 68 human breast...
Monoclonal antibodies (mAB) against progesterone receptor (PR) and the peroxidase-antiperoxidase (PAP) method to visualize PR in paraffin sections from 68 human breast cancers were used. Ten mAB, which recognize human PR on frozen sections, were tested. Six could detect PR in paraffin sections, with Li 417, LET 456, and LET 126 giving the best results. LET 126 antibody was used for most further studies. The effects of fixation with picric acid-formaldehyde (PAF), buffered-formalin, or with Bouin solution were investigated; all fixation methods allowed PR immunolabeling, although PAF or buffered formalin usually gave the best results. Positive staining was seen in the nucleus of carcinoma cells. Variations in intensity and extent of immunoreactivity were observed in all sections and among different regions of the same specimen. These were probably related to the heterogeneity of the tumor cell population. Results were compared with the PR content in the respective tumor tissues, determined by steroid-binding assay, and with immunocytochemistry on frozen sections. It was shown that there were correlations between the immunocytochemical staining (positive or negative) and the steroid binding assay (80%) and between the immunocytochemical staining on paraffin sections and on frozen sections (78%). Weaker intensity and fewer number of PR-positive cells were found for paraffin-embedded tumors. Estrogen receptors were also detected on adjacent sections from the same paraffin-embedded tissues by use of monoclonal anti-ER antibodies (ERICA-kit[Abbott Laboratories, Chicago, IL]) and DNase pretreatment. In conclusion, this immunocytochemical method for detection of PR and ER on paraffin sections offers an alternative to frozen tissue. It allows histologic and immunocytochemical studies on the same sample and retrospective studies on stored tissue blocks.
Topics: Antibodies, Monoclonal; Breast Neoplasms; Frozen Sections; Humans; Immunoenzyme Techniques; Paraffin; Receptors, Estrogen; Receptors, Progesterone
PubMed: 2675623
DOI: No ID Found