Aplicación actual de la clasificación Inmunohistoquímica de cáncer de mama / Current Applicaton of Immunohistochemistry in Breast Cancer

Introducción: El cáncer de mama es la primera causa de mortalidad en el mundo y en Guatemala ocupa el segundo lugar en frecuencia. Desde 1991 su clasifcación ha evolucionado a fn de mejorar el pronóstco y su tratamiento basado en la descripción del tpo y grado histológico. El objetvo de este estudio es conocer la aplicación actual de la inmunohistoquímica en cancer de mama. Metodología: Estudio observacional descriptvo retrospectvo, sobre cáncer de mama clasifcado por inmunohistoquímica en 281 pacientes en el Insttuto Guatemalteco de Seguridad Social (IGSS) de enero 2012 a enero del 2017. Resultados: El subtpo Luminal A se presentó en el 31% de los pacientes y su tratamiento fue principalmente hormonal; seguido por el Triple Negatvo en el 26% tratado mayoritariamente con quimioterapia. El HER2 Positvo en el 21%, el subtpo B-like 13% y en menor frecuencia el Luminal B en el 9% de los casos. El subtpo triple negatvo presento mayor recurrencia y mortalidad con mayor elevación del P53 (40%, OR 2.4) y Ki67 (37%, OR 1.4). La edad en la mayoría de pacientes incluidas fueron superiores a los 40 años y los estadios II y III se presentaron con mayor frecuencia. Conclusiones: El estudio de la inmunohistoquímica realizado en los tejidos obtenidos permitó mostrar que el subtpo luminal A fue el más frecuente, recibiendo en la mayoría de los casos tratamiento hormonal. El más agresivo en recurrencia, metástasis, mortalidad y con valor elevado del Gen p53 y Ki67, fue el Triple Negatvo, tratado con quimioterapia.

Background: Breast cancer is the leading cause of death in the world, and in Guatemala it is the second most common cause of death. Since 1991, its classifcaton has evolved in order to improve prognosis and treatment based on the descripton of type and histological grade. The purpose of this study is to learn the current applicaton of immunohistochemistry of breast cancer. Methods: This retrospectve descriptve observatonal study of breast cancer classifed by immunohistochemistry was performed on 281 patents at the Guatemalan Social Security Insttute (IGSS) between January 2012 and January 2017. Results: The subtype Luminal A was present in 31% of the patents and treatment was mainly hormonal; followed by Triple Negatve in 26% of patents treated mainly with chemotherapy. HER2 positve in 21% of patents, subtype B-like in 13% and lower frequency Luminal B in 9% of the patents. The triple negatve subtype presented higher recurrence and mortality with P53 elevaton (40%, OR 2.4) and Ki67 (37%, OR 1.4). Average age in the study was 40 years old and stages II and III were more frequent. Conclusions: The study of immunohistochemistry performed on tssue obtained, demonstrated that Luminal A subtype is the most frequent, in which the majority of patents received hormonal treatment. The most aggressive in recurrence, metastasis, mortality and with high values of gene p53 and Ki67, was the Triple Negatve, which were treated with chemotherapy.

Usefulness of Immunohistochemistry for Microsatellite Instability Screening in Gastric Cancer

BACKGROUND/AIMS: The usefulness of immunohistochemistry to screen for the microsatellite instability (MSI) phenotype in gastric cancer remains unclear. Moreover, the prognostic value of MSI phenotypes in gastric cancer has been debated. METHODS: The clinicopathologic parameters and survival outcomes of 203 MSI-high (MSI-H) and 261 microsatellite-stable (MSS) advanced gastric cancers (AGCs) were compared. Next, we compared the immunohistochemistry results for hMLH1 and hMSH2 with those of a polymerase chain reaction (PCR)-based method. Kaplan-Meier curves and a Cox proportional hazard regression model were used to conduct survival analyses. RESULTS: The MSI-H AGCs were correlated with older age (p<0.001), female gender (p=0.018), distal location (p<0.001), larger size (p=0.016), and intestinal type (p<0.001). Multivariate analysis revealed that the MSI-H phenotype was an independent favorable factor that was related to overall survival in patients with AGC (p<0.001). Compared with the PCR-based analysis, immunohistochemistry exhibited high sensitivity (91.1%) and specificity (98.5%) in the detection of MSI phenotypes. CONCLUSIONS: MSI-H gastric cancers have distinct clinicopathologic features and better prognoses, which suggests the necessity of MSI analysis in gastric cancer. Immunohistochemistry can be a useful and reliable screening method in the assessment of MSI status in gastric cancer.

Immunohistochemical expression of nuclear factor kappa-B/p65 and cyclooxygenase-2 in non-small cell lung cancer patients: Prognostic value and impact on survival

Nuclear factor kappa B/p65 [NF-KB/p65] regulates the expression of various molecules important in tumorigenesis as cyclooxygenase-2 [COX-2]. To study the immunohistochemical expression of NF-KB/p65 and COX-2 in NSCLC, investigate the relationship between their expression and the clinicopathological parameters, evaluate their prognostic value and clarify their impact on survival. Fifty NSCLC patients were enrolled in this study and subjected to: full medical history and physical examination, routinelaboratory tests and CT chest. Fiberoptic bronchos-copy was done and biopsies were taken from visible masses and 20 mucosal biopsies of the same patients [as control]; sent for histopathological and immunohistochemical examination using anti-COX2 and anti-NF-KB/p65 antibodies. The median follow up of patients was 13 [range 4-22 months]. Results: Thirty six [72%] showed positive NF-icB/p65 expression, it was higher in squamous cell carcinoma and adenocarcinoma than in normal biopsies. Adenocarcinoma showed higher NF-KB/ p65 positivity compared to squamous and large cell carcinomas. A significant relationship was found between NF-KB/p65 expression and overall survival. Forty five [90%] showed positive COX-2 expression, no expression was detected in normal biopsies. COX-2 expression was significantly higher in adenocarcinoma compared to squamous and large cell carcinomas. NF-KB/p65 and COX-2 expression was significantly correlated with advanced tumor stage, lymph node metastasis and grade. A significant positive relationship was found between NF-KB/p65 and COX-2 expression. NF-KB/P65 and COX-2 expression has a prognostic value in NSCLC, they are suitable targets for development of new lung cancer therapies; inhibition of NF-KB and COX-2 will augment the efficacy of anticancer therapy

Histological and immunohistochemical study on the survival of diced cartilage grafts wrapped in surgical and fascia in albino rabbits

Diced cartilage graft is commonly used in plastic and reconstructive surgery such as auricular reconstruction and rhinoplasty. Wrapping diced cartilage with fascia or surgicel or other wrappers can affect the survival of cartilage grafts. This study was carried out to verify the survival of cartilage grafts when they are unwrapped or wrapped with surgicel versus fascia. This study was carried out on 20 adult male albino rabbits. They were divided into four equal groups. Cartilage grafts were harvested from both ears of group I. One part was used as the control and the other part was diced into 0.5-mm pieces and implanted into subcutaneous dorsal pockets of the other three groups as follows: group II received unwrapped diced cartilage grafts, group III received diced cartilage graft wrapped in surgicel, and group IV received diced cartilage graft wrapped in fascia. At the end of the eighth week, the rabbits were sacrificed and the original surgical sites were incised with their implants. The specimens were prepared for histological and immunohistochemical study using glial fibrillary acidic protein to be examined by means of a light microscope. Cartilage grafts wrapped in fascia showed the best survival, and cell viability was more or less similar to that of normal cartilage. They showed significant increase in the nuclear lacunar count, number of glial fibrillary acidic protein -positive chondrocytes, and in the pixel intensity of Mallory trichrome-stained sections. This was superior to those kept without wrapping and to those wrapped in surgicel. Diced cartilage grafts wrapped with fascia demonstrated normal cartilage architecture with respect to chondrocyte viability and collagen and elastic content compared with cartilage grafts kept without wrapping or those wrapped in surgicel

Histological and immunohistochemical study on the possible protective effect of curcumin on intestinal ischemia / reperfusion induced lung injury in albino rats

Intestinal ischemia-reperfusion [I/R] injury may lead to severe local and remote tissue injury, especially acute lung injury. Curcumin has been shown to attenuate [I/R] injury in several organ systems. To evaluate the possible protective role of curcumin on acute lung injury induced by intestinal [I/R] in albino rat. Thirty male albino rats were divided equally into control group [Sham-operated], which underwent laparotomy without superior mesenteric artery occlusion. It was classified into two subgroups: subgroup a: which received no treatment, subgroup b: which received curcumin, I/R group where rats were exposed to superior mesenteric artery occlusion for 1 hour followed by 1 hour reperfusion. Third group was I/R and curcumin. Curcumin, was administered in a dose of [100 mg/kg] daily orally for 15 days before experiment. Rats were sacrificed at the end of reperfusion. Lung sections were stained with H and E and Cyclooxygenase -2 immunostain. A morphometric study and statistical analysis were performed. Intestinal I/R caused severe histological changes including massive infiltration of cells in thickened alveolar septa, around bronchioles and congested blood vessels. Curcumin led to obvious regression of induced histological changes by I/R. Significant increase in the mean thickness of inter alveolar septa and the mean area% of Cox-2 immunoexpression were recorded in [I/R] group compared to other groups. Curcumin administration proved a beneficial protective effect on I/R induced acute lung injury, and therefore has potential for clinical use

Effect of bisphenol-A on the lung of adult male albino rats and the possible protective role of geraniol: a histological and immunohistochemiacal study

Concern is mounting regarding the human health and environmental effects of bisphenol A [BPA], a hormone-disrupting chemical used in plastic and epoxy resin industries. Geraniol [a plant-derived terpene] has cytoprotective and antioxidant potential. This work aimed to study the effect of BPA on the lung of adult male albino rats, assess different dose responses of BPA, and evaluate the possible protective role of geraniol supplementation using histological and immunohistochemical methods. Thirty-five adult male albino rats were divided into seven equal groups. All treatments were given by oral gavage for 8 weeks. Group I served as the untreated control group. Group II received corn oil [0.5 ml/day]. Group III was given geraniol [150 mg/kg body weight/on alternate days]. Groups IV, V, and VI received BPA [5, 50, and 500 mg/kg body weight/day, respectively]. Group VII was treated with BPA [500 mg/kg body weight/day] along with geraniol at a similar dose. Lung specimens were taken and processed for H and E, Masson's trichrome stains, and for immunohistochemical staining of alveolar macrophages [CD68] and inducible nitric oxide synthase [iNOS]. The area percentage of positive iNOS immunoreactivity and the count of alveolar macrophages in immunostained sections were morphometrically and statistically analyzed. Group IV revealed similar results to the control group. Groups V and VI showed disrupted lung architecture with collapsed alveoli, inflammatory cellular infiltration, congested thickened pulmonary vessels, extravasated red blood cells, and collagen fiber deposition. There was a significant increase in the area percentage of positive iNOS immunoreaction and in the count of alveolar macrophages, compared with the control group. These effects were more pronounced in group VI. Coadminstration of BPA and geraniol [group VII] preserved a near-normal lung structure. Long-term administration of BPA caused dose-dependent lung damage, which could be significantly ameliorated by concomitant treatment with geraniol

Quantitative evaluation of pulomnary neuroendocrine cells and the role of nitric oxide in experimentally induced renovascular hypertension in male albino rats

Renovascular hypertension is the most common cause of secondary hypertension that leads to disorders in many systems and organs including the lung. The present work was carried out to determine how experimentally induced renovascular hypertension affects rat lung tissue with a special focus on the disorders in nitric oxide [NO] production as well as the number of pulmonary neuroendocrine cells. Twenty-one adult male albino rats were used; they were divided into three groups. Group I included five control rats, group II included eight rats that were sham operated, and group III included eight rats that were subjected to a renovascular hypertension procedure. Lower segments of the right lung lobes were obtained and then subjected to H and E and Masson's trichrome stains. Endothelial nitric oxide synthase [eNOS] and calcitonin antibody-2 were also used as immunohistochemical stains. Morphometric measurements included the area% of collagen fibers, the optical density of eNOS immunoreaction, and the number of calcitonin-positive neuroendocrine cells. Data were recorded and analyzed statistically. In group III, H and E-stained sections showed thickening of the tunica media of pulmonary arteries and alveolar walls with predominantly type II pneumocytes. Consequently, the thickened arteries showed a significant increase in the mean area% of collagen fibers, together with a significant decrease in the mean optical density of eNOS immunoreactivity when compared with the control group. The mean number of calcitonin immunoreactive neuroendocrine cells of group III showed a significant increase versus the control group. The current study showed an increased number of pulmonary neuroendocrine cells with renovascular hypertension. This increase may represent a compensatory mechanism to overcome renovascular hypertension. Thus, clinical trials could be conducted to prove the possible role of calcitonin-producing pulmonary neuroendocrine cells in controlling renovascular hypertension

histological changes of the female rat mammary gland during the fertile period with a special reference to E-cadherin expression

E-cadherin [E-cad] is a cell adhesion molecule, essential for the maintenance of the epithelial phenotype. The current study aimed at showing the histological changes and the pattern of E-cad expression in the female rat mammary gland during different stages of their fertile period microscopically and morphometrically. Thirty-five adult female albino rats [3-4 months old] were divided into seven groups [five rats each]: the rats in group I were not pregnant [resting] and groups II-VII were sacrificed at days 7 and 14 of each of their pregnancy, lactation, and weaning stages. The excised right inguinal mammary glands were processed for H and E and E-cad immunostaining. The sections were evaluated microscopically and morphometrically. The mammary gland of nonpregnant rats had abundant adipose connective tissue with few glandular elements. During pregnancy, there was a progressive increase in the alveoli, which became distended with milk, with prominent sac formation during lactation. The E-cad expression pattern was mainly observed as a strong positive linear membranous pattern in all borders of epithelial cell lining, whereas the myoepithelial cells were negative. These observations were found in different groups, except that of lactation, where the pattern was limited to the basolateral membranes of these epithelial cells. Morphometrically, the positive membranous epithelial cells for E-cad, its density area fraction and cytoplasmic positive reaction ranged from 7-83%, 3.9-6.8% and 0-7%, respectively in all groups. In late pregnancy, there was a marked increase in the negative epithelial cells [93%], with a significant decrease in density area fraction expression [3.9%], which was persistent during lactation. Positive E-cad expression of stromal cells was only observed during weaning. The E-cad was mainly localized to the membranes of epithelial cell lining and absent in myoepithelial cells. The quantitative E-cad expression can represent a tool that ensures qualitative microscopic observations during the fertile period

Effect of induced diabetes mellius on the histological structure of the renal cortex of adult male albino rats and the possible protective role of licorice extract

Diabetes is one of the most common chronic metabolic disorders worldwide. One of the major complications of type 2 diabetes is diabetic nephropathy. The present study was to investigate the effect of type 2 diabetes on the histological structure of the renal cortex of adult male albino rats and the role of licorice ethanolic extract on diabetic renal affection. Forty adult male albino rats were utilized. They were classified into three main groups: the control group [group I], the experimental diabetic group [group II], and the possible protected group [group III]. Type 2 diabetes was induced in rats in groups II and III by giving them a high-fat diet and a single low dose of streptozotocin. Diabetic rats were divided into two subgroups: untreated subgroup IIa and treated subgroup IIb. The possible protected group received licorice ethanolic extract concomitant with the high-fat diet and the single low dose of streptozotocin. At the end of the experiment, the kidneys were dissected out and processed for light and electron microscopic examination. Fasting blood glucose level, fasting insulin level, serum urea, and creatinine were estimated and statistically analyzed. Examination of the renal cortex of untreated diabetic subgroup IIa demonstrated glomerulosclerosis and distorted podocyte foot processes. The cells lining convoluted tubules revealed thick basement membranes, disorganization of basal infoldings, and mitochondrial disarrangement. The area% of positive Bax immunoreaction was significantly increased in subgroup IIa as compared with subgroup IIb and group III. Examination of the renal cortex of the treated diabetic animals [subgroup IIb] revealed little improvement and failure of licorice extract to normalize renal cortical changes, most probably due to late intervention. In contrast, the protected group [group III] revealed a nearly preserved normal architecture. Changes in the renal cortical structure were attenuated with prophylactic therapy of licorice ethanolic extract

histological and immunohistochemical study of the effects of leptin on the pars distalis of the pituitary gland in female albino rats

Leptin is an adipocyte-derived hormone. The plasma level of leptin is elevated in obesity and decreased in emaciation. It is involved in the neuroendocrine regulation of pituitary gland function. The present work was carried out to study the histological changes in the pars distalis of the pituitary gland after an injection of leptin in female albino rats. Thirty just-weaned immature female albino rats [22 days] were divided into two groups: group I included 10 animals and served as a control group and group II included 20 animals that received a daily subcutaneous injection of 5 micro g leptin up to the end of the experiment. The females were subjected daily to vaginal smear. After the maturation was established, rats from each group were again subdivided into groups a and b, which were sacrificed 4 and 20 days after puberty, respectively. The pituitary glands were dissected and specimens were prepared for electron microscopic and immunohistochemical assessment. Leptin injection induced various changes in the pars distalis of the pituitary gland. Some cells had deeply stained nuclei and vacuolated cytoplasm. There was a statistically significant increase in the number of positive p53 antibody immunostained cells in the leptin-treated group, especially after 4 days of leptin injection. The most affected cells were somatotrophs, thyrotrophs, and gonadotrophs. They showed features of hyperactivity, with the appearance of some apoptotic cells. Later, necrotic changes such as pyknotic nuclei, ballooned mitochondria with destroyed cristae, and dilated rough endoplasmic reticulum were observed. Prepubertal increase in leptin led to histological changes in some cells of the pars distalis. It is recommended to avoid increase in body weight, especially at a young age

Oviductal isthmus light microscopic changes induced by clomiphene citrate in albino rats

Clomiphene citrate [CC] therapy for cases of infertility has been linked to increased incidence of tubal ectopic pregnancy. This study aimed to investigate the histological changes produced by CC in the isthmus portion of albino rat oviducts. Twenty adult female albino rats were divided into three groups. The first was the control group. The second group received 1 mg/kg/day CC for 4 days, whereas the third group was allowed a recovery period after receiving CC. The oviductal isthmus sections were stained by H and E, periodic acid-Schiff [PAS], and caspase for light microscopic examinations. An image analyzer was used and the measured data were recorded and statistically analyzed. The CC intake led to increased thickness of the oviductal isthmus lining epithelium, as well as to hyperplasia, bridging, and reduction of the lumen diameter. Furthermore, many degenerative and apoptotic changes as well as significant increase in the PAS optical density and caspase reaction area percentage of the lining epithelium were detected. Also, the isthmic sections of the recovery group revealed increased wall thickness, epithelial degenerative changes, and hyperplasia, in addition to significant reduction in the lumen diameter leading to tubal obstruction. Significant increase in the PAS optical density of the lining epithelium was detected, but its caspase reaction area percentage showed insignificant change compared with the control group sections. This study suggests that CC administration for 4 days at a dose of 1 mg/kg/day induces damage to the albino rat oviductal isthmus that could predispose to occurrence of ectopic pregnancy and that progressively developed despite stoppage of the drug

Light and electron microscopic study on the effect of diazepam on the cardiac muscle of adult albino rat and the possible role of garlic

Diazepam is a drug that belongs to a group of pharmacological agents called benzodiazepines. Several studies have indicated that diazepam exerts a myocardial depressant effect. Garlic preparations have been used widely for the prevention and treatment of cardiovascular conditions. This work aimed to study the effect of diazepam on the cardiac muscle of adult albino rat and the possible role of garlic as a protective agent. Forty adult male albino rats were divided into four equal groups: control group, garlic-treated group [250mg/kg body weight], diazepam-treated group [1mg/kg body weight], and both garlic and diazepam-treated group. The animals were treated orally daily for 4 weeks. Specimens from the cardiac muscle were processed for light and electron microscopy. Immunohistochemical study was carried out using an antibody against vimentin. Specimens from diazepam-treated animals showed focal disruption of cardiac myocytes, peripheral deeply stained nuclei, and vacuolated sarcoplasm. Focal aggregation of mononuclear cells and dilated congested blood vessels were also observed in between the myocytes. Ultrastructurally, irregular indented nuclei, focal lysis of the myofibrils, loss of normal cross striations, swelling of mitochondria, and distortion of intercalated disks were also observed. Immunohistochemical study showed a highly significant increase in vimentin immunoreaction in the endomysial and perimysial sheaths, in the walls of blood vessels, and in some interstitial cells. In contrast, minimal changes were observed in rats treated concomitantly with both garlic and diazepam, with a non significant increase in the immunoreaction. Diazepam induced structural changes in rat cardiac muscle that could be ameliorated by concomitant treatment with garlic

possible ameliorating effect of Nigella sativa oil on tramadol induced apoptosis in the motor area of rat cerebral cortex: a histological and immunohistochemical study

Tramadol is a centrally active analgesic commonly prescribed for moderate to severe pain. Thymoquinone, the major active component of the Nigella sativa oil, is characterized by its antioxidant properties. This study aimed to demonstrate the histological and p53-immunohistochemical changes induced by tramadol in the rat cerebral cortex and evaluate the potential role of N. sativa oil in the attenuation of these changes. Twenty-four male albino rats divided into three groups were used in this study. Group I was the control group. Group II was given repeated intraperitoneal injections of increasing doses of tramadol of 20, 40, and 80 mg/kg/day on the first, second, and third ten days of the study, respectively. Group III was given oral N. sativa oil 4 ml/kg/day, 30 min before each tramadol injection for 30 days. Paraffin sections of the frontal cortex motor area were prepared and stained with H and E and with an immunohistochemical stain using anti-p53 antibody. In group II rats, numerous shrunken pyramidal cells with acidophilic cytoplasm and deeply stained pyknotic nuclei were seen. Some of the granular cells appeared as ghosts with margination of chromatin. Homogeneous acidophilic masses containing fragmented deeply stained nuclei and surrounded by clear halos were also observed. The number of p53-positive cells was significantly higher compared with both group I and group III. In contrast, in group III, multiple pyramidal and granular cells appeared normal and the number of p53-positive cells was significantly less compared with group II. N. sativa oil and derived thymoquinone ameliorate tramadol-induced apoptosis in the motor area of the rat cerebral cortex

Can gold nanoparticles affect the histological structure of the pulmonary alveoli in adult albino rats?

Gold nanoparticles [GNPs] are currently used in several consumer products as well as in laboratory-based research either as a diagnostic imaging agent or as a therapeutic agent in gene delivery and photothermal cancer therapy. They are synthesized by chemical methods in which toxic reducing agents are involved. There is a growing need to develop environmentally benign NP synthesis processes that are free from toxic chemicals. The study aimed to ascertain the possible histological alterations that might occur in the pulmonary alveoli of adult albino rats after intraperitoneal administration of 10 nm GNPs prepared by two different methods [chemically prepared and green synthesized]. Fifty adult male albino rats [140-160 g] were assigned to three groups. Group I was the control group. The rats in this group were further divided into three equal subgroups Ia, Ib, and Ic that received daily intraperitoneal injection of PBS, trisodium citrate, and cumin-gum Arabic solutions, respectively. Rats in group II received a daily intraperitoneal injection of 100 micro l of chemically prepared GNPs using trisodium citrate. Rats in group III received a daily intraperitoneal injection of 100 micro l of green-synthesized GNPs using cumin and gum Arabic. After 7 days, the animals were euthanized and specimens from the lungs were taken and processed for histological and immunohistochemical studies. H and E-stained sections of group II rats depicted multifocal thickening of the interalveolar septa and collapsed alveoli. Immunohistochemical analysis revealed high vimentin expression. Ultrastructurally, type II pneumocytes showed degenerative changes. Relatively thick interalveolar septa with many active interstitial cells were also seen. GNPs were detected in pneumocytes and alveolar macrophages. In contrast, the lung sections of group III rats showed considerable preservation of normal alveolar architecture with nearly normal vimentin immunoreactivity. GNPs were detected in the pneumocytes and interalveolar septa as well. Green-synthesized GNPs using cumin-gum Arabic are safer for the pulmonary alveoli compared with chemically prepared GNPs using trisodium citrate. Long-term studies are required to verify the toxicity of GNPs on various cell types

histological and immunohistochemical study on the possible protective effect of omega-3 polyunsaturated fatty acids on isoprenaline induced myocardial fibrosis in rats

Preclinical and clinical studies have demonstrated that omega-3-polyunsaturated fatty acids [omega-3-PUFAs] play a significant role in the prevention of cardiovascular diseases. This study aimed to investigate the possible protective effect of omega-3-PUFAs on isoprenaline [ISP]-induced myocardial fibrosis. Thirty-two adult male albino rats were divided into five groups. Group I represented the control group [eight rats]. In group II, six rats were given omega-3-PUFAs [40 mg/kg/day] orally for 8 weeks. In group III, six rats were injected subcutaneously with ISP [5 mg/kg/day] for 4 consecutive days and sacrificed 2 days later. In group IV, six rats were injected with ISP [5 mg/kg/day] for 4 consecutive days and sacrificed after 2 weeks. In group V, six rats were given omega-3-PUFAs [40 mg/kg/day] for 8 weeks, following which they were injected with ISP [5 mg/kg/day] for 4 consecutive days, and sacrificed 2 days later. Serum creatine phosphokinase-MB [CPK-MB] was measured. Myocardial sections were subjected to H and E, Masson's trichrome stain, and alpha-smooth muscle actin [alpha-SMA] immunohistochemical stain. Group II showed nonsignificant difference in the mean CPK-MB level compared with the control. Myocardial sections revealed a histological architecture similar to that of the control. The mean area% of collagen and alpha-SMA immunoreactivity was nonsignificant when compared with the control. Group III showed significant increase in mean CPK-MB compared with the control. Myocardial sections showed disorganization, inflammation, exudation, and fibrosis with significant increase in the mean area% of collagen and alpha-SMA immunoreactivity compared with the control. Group IV showed significant increase in mean CPK-MB, area% of collagen, and alpha-SMA immunoreactivity compared with the control, with progression in the myocardial histological alterations. Group V showed significant decrease in the mean CPK-MB with decrease in histological changes, and there was significant decrease in the mean area% of collagen and alpha-SMA immunoreactivity compared with the ISP groups. omega-3-PUFAs exert cardioprotective effects against ISP-induced myocardial fibrosis

Histological study of the effect of bone marrow-derived mesenchymal stem cells on healing of skin defect in adult male albino rats

Despite advances in wound care, skin loss results in significant morbidity and mortality. There is increasing evidence that bone marrow-derived mesenchymal stem cells [BM-MSCs] are useful for tissue regeneration because of their multipotency and easy isolation and culture. The objective of this study was to evaluate the efficacy of local administration of in-vitro-expanded BM-MSCs in the healing of experimentally induced full-thickness excisional wounds in adult male albino rats. Thirty-five adult male albino rats were used. They were divided into three groups. In group I rats, BM was isolated and cultured and skin specimens were obtained as a control. In group II rats, a full-thickness circular skin wound of 5mm in diameter was inflicted on the mid back of each rat and was examined on days 3, 7, and 14. In group III rats, the wound was inflicted as in group II, which was then treated with BM-MSCs and examined as in group II. The wound areas were excised and used for histological and immunohistochemical studies for CD105. Morphometric analysis was performed for assessment of epidermal thickness, area percentage of collagen and elastic fibers, and number of CD105-positive cells. Wounds treated with BM-MSCs [group III] showed evidence of re-epithelialization, increased epidermal thickness, hair follicle formation, collagen, and elastic fibers compared with wounds in group II. Similarly, the number of CD105-positive cells was prominently increased in the skin of the same group. Local administration of in-vitro-expanded BM-MSCs accelerates and promotes healing in full-thickness excisional wounds

possible protective role of selenium supplementation on the structure of trhe heart in an experimental model of chronic renal failure: a light and electron microscopic study

Chronic kidney disease [CKD] is a worldwide health hazard with significant morbidity and mortality especially due to cardiovascular complications. This study was conducted to investigate the histological effects of CKD on the structure of the heart and to show the possible protective role of selenium [Se] supplements. Twenty adult male Wistar albino rats were used in this study. They were divided into three groups: group I [the control group] was divided into two subgroups: IA [sham operated], which was subjected to sham operation, and IB [the negative control group], which was left untreated; group II [the nephrectomy group] was subjected to a five-sixth nephrectomy operation to induce renal failure; group III [the Se-treated group] was subjected to a five/sixth nephrectomy operation following which the rats were supplemented with sodium selenite at a dose of 0.04 mg/kg for 10 weeks. At the end of the experiment body weight [BW] and serum creatinine were measured. Heart specimens were processed for light and electron microscopic examination, and morphometric analysis was performed for area percentage of intercellular spaces and collagen fibers. Group II showed significant widening of the interfiber spaces with aggregation of adipocytes, extravasated RBCs, and lymphocytes. There was also significant increase in the area percentage of collagen fibers. Cardiac myocytes appeared swollen, pale, and degenerated, and electron microscopic findings pointed to endothelial dysfunction. Se administration led to significant improvement in BW, creatinine level, and cardiac fibrosis; yet, the heart showed focal pale vacuolated myocytes, wide interfiber spaces, and extravasated blood. CKD led to definite focal degeneration in the cardiac muscle fibers. Se with the dose given improved BW, creatinine level, and cardiac fibrosis but could not offer complete protection against cardiovascular complication

histological study on the effect of pentoxifylline on acute lung injury induced by renal ischemia reperfusion

Kidney ischemia-reperfusion injury [IRI] occurs under many clinical conditions. It leads to acute kidney injury, which may affect various remote but important organs such as the lung. The aim of the present study was to investigate the effect of renal IRI on the lung structure and evaluate the possible protective effect of pentoxifylline [PTX]. Fifteen adult male Wistar rats aged 8-10 weeks and weighing 150-200 g were used in this study. They were divided into the following groups: group I [the control group] was subjected to sham operation; group II [the IRI group] was subjected to renal ischemia by bilateral clamping of the renal pedicles for 45 min and then allowed 24 h of reperfusion. Group III [the PTX-treated group] was given two doses of PTX [40 mg/kg] intraperitoneally and subjected to an ischemia-reperfusion procedure as performed in group II. Lung specimens were processed for light and electron microscopic examination. The mean thickness of the interalveolar septa and the mean number of type-II pneumocytes were measured. Group II showed diffuse lung injury affecting mainly the alveoli, which appeared flooded with exudate and red blood cells, which also extravasated in the interstitium. There was vacuolation of the cytoplasm of both type-I and type-II pneumocytes with significant increase in the number of type-II cells and depletion of lamellar bodies. Significant increase in the mean thickness of the interalveolar septa was detected and it showed infiltration by macrophages and neutrophils. PTX caused marked improvement in the lung structure after ischemia-reperfusion. Most of the alveoli appeared empty and a few of them showed minimal red blood cells and macrophages. Thin interalveolar septa were detected with thickness comparable to that of the control group. Renal IRI caused alteration in the lung structure, which was ameliorated by PTX administration

Effect of high salt loading on the retina of adult male rats with special reference to aquaporin 1: a histological and immunohistochemical study

In the retina, glial cells control ionic concentrations by mediation of transmembrane water fluxes through aquaporin [AQP] water channels. The risk factor of a high-salt diet on renal and cardiovascular systems is pretty well known. However, it is not yet known whether a high-salt diet alone can affect the retina. The aim of this study was to determine whether a high-salt diet alone can induce changes in the retina and whether it may be accompanied by changes in the expression and immunolocalization of water channel aquaporin1 [AQP1]. Forty-two adult male albino rats were used. They were divided into three equal groups. Group I served as the control group. Rats in group II were administered 2 ml of a high-salt solution [8% NaCl concentration] once daily by means of a gastric tube. Group III was the recovery group. Retinal tissues were collected and examined by means of light and electron microscopy. Immunohistochemical analysis using AQP1 and glial fibrillary acidic protein [GFAP] antibodies was performed and the results were statistically analyzed. The retina of rats given a high-salt diet [group II] displayed obvious disorganization of the outer segment of photoreceptors, together with cytoplasmic vacuolations in the cells of the inner nuclear and ganglionic layers. Furthermore, significant increase in AQP1 and GFAP immunoexpression was detected. In the recovery group [group III] the retinae of some rats regained their normal histological appearance, whereas others failed to do so. High salt loading might alter glial cell-mediated water transport through AQP1 channels in the retina

Histological and immunohistochemical study of nitric oxide synthase and effects of angiotensin receptor blockade in early phase of diabetes in rat kidney

Several studies have demonstrated that the pathophysiological and morphological changes in early diabetic nephropathy were mediated by an increase or decrease in nitric oxide [NO] production and/or activity. There are few reports suggesting a relationship between NO and the renin-angiotensin system. The present study was designed to determine the effects of early diabetic state on NO production and also to assess the possible effects of angiotensin receptor blockers [ARBs] on these changes. Thirty adult male albino rats were included in this study. Twenty were injected with streptozotocin for induction of diabetes. The other 10 were injected with the vehicle and served as control. Two days after injection, the diabetic animals were randomly divided into two groups of 10 animals each. One group was given valsartan as an ARB and the other group received no further treatment. Three weeks later, all animals were sacrificed and the kidneys were processed for obtaining paraffin sections. The sections were stained with H and E, Masson's trichrome, and periodic acid-Schiff. The sections were also stained with an immunohistochemical stain against endothelium-derived nitric oxide synthase [eNOS]. Diabetes induced histological changes in the form of glomerular hypertrophy, increased glomerular matrix, focal areas of tubular atrophy, medullary congestion, and slight fibrosis. Immunostaining was present in the control kidney in the glomeruli and in the collecting tubules of the medulla. Diabetes induced a positive reaction in the proximal and distal convoluted tubules and increased immunoreactivity in the collecting tubules. Treatment with valsartan resulted in an improvement in the morphology of the kidney and a reduction in the intensity of eNOS immunostaining. NO increases in early diabetic kidney and ARB in the form of valsartan could be recommended for preventing the development of diabetic nephropathy