Effect of Stem Cell Therapy on Induced Diabetic Keratopathy in Albino Rat

BACKGROUND AND OBJECTIVES: Type 2 diabetes mellitus (DM) is a prevalent disorder. Diabetic keratopathy is a well-known ocular complication secondary to type 2 DM. Topical insulin application did not affect apoptosis and necrosis levels in corneal epithelium. Autologous cell transplantation is not a viable option for diabetic patients with bilateral limbal stem cell deficiency. The present study aimed at assessing the possible effect of hemopoeitic stem cell (HSC) therapy on induced diabetic keratopathy in albino rat. METHODS AND RESULTS: Fifteen male albino rats were divided into control group of 2 rats, diabetic group of 8 rats receiving single intraperitoneal (IP) injection of 50 mg/kg streptozotocin (STZ). 3 animals were sacrificed 6 weeks following confirmation of diabetes to confirm keratopathy and 5 rats were sacrificed 4 weeks following confirmation of keratopathy. SC therapy group included 5 rats injected with HSCs 6 weeks following confirmation of diabetes and sacrificed 4 weeks following SC therapy. Cord blood collection, stem cells isolation and labeling were performed. Eye specimens were subjected to histological, histochemical, immunohistochemical, morphometric and statistical studies. In diabetic group, the central cornea showed multiple cells with vacuolated cytoplasm and dark nuclei, focal epithelial discontinuity, reduced corneal thickness and less number of layers of corneal and conjunctival epithelia. In stem cell therapy group, few cells with vacuolated cytoplasm and dark nuclei were found in the corneal and conjunctival epithelia with more number of epithelial layers. CONCLUSIONS: A definite ameliorating effect of HSC therapy was detected on diabetic keratopathy. The therapeutic cells were effective in limiting corneal epithelial changes.

Reproductive effects of human interferon-Alpha-2b administration on male albino mice testes. An experimental study

Background: recombinant human interferon alpha [rh-IFN-alpha] is used therapeutically in malignant disorders and chronic hepatitis. The phenotypic effects of this drug at the structural levels on testicular tissue were hardly ever addressed. Hence, this work was designed in adult male albino mice to study the phenotypic effects of rh-INF-alpha-2b on testicular tissue as well as assessing its effects on serum testosterone and gonadotropins levels

Objective: this research was planned to through light on the effects of interferon-alpha-2b [IFN-alpha-2b] on the hypothalamic-pituitary-testicular [HPT] axis of the adult male albino mice

Design: experimental study

Setting: national Hepatology and Tropical Medicine Research Institute [NHTMRI]. The study was conducted from November [2004] to February [2005]

Materials and methods: thirty sexually mature male mice were divided into three groups [10 mice in each group], namely: the control, the experimental and the recovery groups. Mice in the experimental and recovery groups were administered recombinant human interferon alpha intraperitoneally at a dose of 3000 U / mouse weekly for 12 weeks in a volume of 1.0-microliter isotonic normal saline, then animals in the recovery group were left to recover for a further period of two months. At the end of the experiment, serum concentrations of gonadotropins and testosterone were measured and then all animals were then sacrificed to study histopathologically the possible effects of interferon on the testicular tissue

Results: rh-IFN-alpha-2b induced remarkable decline in the serum levels of both follicle stimulating hormone [FSH] and luteinizing hormone [LH] in mice of the experimental group compared to the corresponding control and mice of recovery group. At the same time, testosterone was moderately increased in the experimental group, and then returned to its normal levels within 2 months after cessation of treatment. Histopathologically, in the experimental group, there were focal thickening of the basement membrane, degenerative changes and clumping of the germinal epithelial cells in the center of seminiferous tubules, partial desquamation of the germinal epithelium from basement membrane, reduction in the germ cell height, partial arrest of maturation and increased number of Sertoli cells. Increased number of Leydig's cells and hypervascularity were detected in the interstitial spaces. In the recovery group, there was lessening of the germ cell hypoplasia manifested by restoration of spermatogenic cells and accidental disruption in the basement membrane. Most of the spermatogenic and Sertoli cells restored their polarity, height and maturation

Conclusion: our results suggest that rh-INF-alpha-2b temporally affects the hypothalamic-pituitary-testicular axis [HPT], both centrally and peripherally [at the testicular level], through the lessening of FSH, LH, raise of testosterone serum levels and direct phenotypic effect on the testicular tissue

Light and electron microscopic study on the effect of training and high cholesterol diet on the skeletal muscle of male albino rat exposed to strenuous exercise

The present study aimed at investigating the effect of training and high cholesterol diet on the skeletal muscle [soleus] of male albino rat exposed to strenuous exercise [SE] immediately and 24 hours after. Eighty eight male albino rats were divided into 2 groups: group 1 given balanced diet and group 2 given high cholesterol diet. Each group was subdivided into control rats not subjected to exercise and experimental group which included untrained and trained rats exposed to [SE] and sacrificed immediately or after 24 hours. Serum triglyceride [TG] and low density lipoprotein cholesterol [LDL-C] were tested. Semithin and ultrathin sections were examined by light and electron microscopes. Mean unclear area and count of sarcoplasmic vacuoles were assessed morphometrically. In group 1, nuclei with peripheral chromatin condensation, darkly stained fibers and electron dense areas were seen in untrained rats immediately following [SE]. Small nuclei with peripheral chromatin condensation, atypically arranged myofibrils, discontinuous A bands, focal loss of Z lines with electron dense particles around and enlarged,mitochondria with destroyed cristae were detected in untrained rat muscles 24 hours following [SE]. Less changes were noticed in corresponding trained rats. Group 2 revealed multiple sarcoplasmic vacuolations. Dark areas, irregular discontinuous myofibrils and small nuclei with peripheral chromatin condensation appeared in untrained rat muscle immediately following [SE]. In addition, small dark nuclei existed 24 hours after [SE]. Highly significant decrease in mean nuclear area and mean count of sarcoplasmic vacuoles existed in untrained and trained subgroups respectively. Dilated cisternae of sarcoplasmic reticulum, enlarged mitochondria with destroyed cristae and irregular focally disrupted sarcolemma were seen in untrained rats. Less changes were detected in corresponding trained rats. Serum [LDL-C] and [TG] showed highly significant increase in group 2 and highly significant decrease in trained subgroups. It was concluded that apoptosis was induced by strenuous exercise exacerbated by hypercholesterolemia and lessened by training

Histological and immunohistochemical study on the rat Kupffer cell in response to short and long term ethanol administration

The present study aimed at investigating the Kupffer cells in the rat liver of both sexes in response to short and long term ethanol administration, as regards multiplicity and CD14 receptor immuno-expression and its relation to the developing damage. Forty eight male and female albino rats were divided into a control group of 12 rats and an experimental group of 36 rats including subgroups I and II [short term ethanol administration] that were given 17.5% ethanol for 1 and 3 days respectively. Subgroup III [long term ethanol administration] was given 34.2% ethanol for 6 weeks. India ink was injected into half the number of male and female control and experimental rats. Liver sections were stained with H. and E. and immunohistochemically using CD14 antibody for the receptors on the surface membrane of Kupffer cells. The count of Kupffer cells, area of CD14 immunoexpression and the optical density of CD14 immunoreaction were assessed. In subgroup II apoptotic cells and in subgroup III hepatocytes exhibiting eosinophilic material and small dark nucleus were found. The changes were in obvious in females. Kupffer cells seen in India ink injected sections revealed nonsignificant difference in their mean count in different groups and subgroups. CD14 immunoexpression appeared as darkly and lightly stained areas in male and female control rats. The immunoexpression and the immunoreaction increased in the experimental subgroups with a remarkable difference between female and male rats. A highly significant increase in the mean area and mean optical density was recorded in subgroup I versus control rats. Also in subgroups II and III versus control rats and rats of subgroup I of the same gender and in experimental female versus male rats. Short and long term ethanol administration led to activation of CD14 receptors on the surface membrane of Kupffer cells, indicating their role in initiation of liver injury

Effect of androgen and schistosomicide in murine schistomiasis mansoni: an experimental study

Background: Impotence is a consistent inability to sustain an erection sufficient for sexual intercourse. Testosterone administration in men with liver cirrhosis improves the sense of well-being, increase serum proteins and reduces edema without serious adverse effects. Oral, alkylated forms of testosterone can create a situation of liver toxicity. There is little evidence that other methods of administration cause liver dysfunction. Most doctors be indecisive on prescribing androgen preparations in patients with liver disease, so this work was designed to study the effect of androgen replacement [injectable form] on the murine diseased liver, and subsequently whether it can be used safely in men with chronic liver disease or not

Objective: To evaluate the effect of exogenous injectable androgen and praziquantel on the diseased liver of mice

Setting: National Hepatology and Tropical Medicine Research Institute [NHTMRI]

Materials and methods: Forty male mouse [weighing 25-30 g] were infested subcutaneously with Schistosoma mansoni [100 cercariae/animal], and then they were divided into four groups. Mice in the first group were infected only and used as infected control group. Mice of group II and IV were given the Schistosomicide, praziquantel in a dose of 0.3mg/mouse. Androgen [Sustanon] was injected intramuscularly in a dose of 0.125 mg/mouse, [three doses, 3 weeks apart] in group III and IV. At the end of the trial all animals were then sacrificed to study histopathologically the possible effects of androgen on the liver tissue. Liver function tests were done in animals of group I, III, and IV, first prior to study and finally by the end of study. Results of assayed liver function tests and histopathological examination were tested for statistical significant association

Results: there were marked elevation of the liver enzymes in mice of group IV compared to the corresponding control [p<0.01] and mice of the third group [p<0.01], which reflect deterioration of hepatic function in those mice received the antibilharzial drug praziquantel. On the other hand there was statistical difference between control group [group I] and androgen treated group III [P < 0.05]. Histological examination of liver sections of mice in all groups revealed the presence of typical bilharzial granulomas. The mean diameter of bilharzial granulomas clearly dropped to 283.20 micrometer in group II compared to 392.55 in corresponding control. The difference between these two groups was statistically significant [p = 0.000].]. In group III there was no statistical difference in the number of egg granulomas [P> 0.05] compared to group I. There was a reduction of granulomas diameter in group III and IV [animals injected with androgen] in comparison to group I [P>0.05 and P<0.01] respectively. Also comparison between the four groups as regards the type of bilharzial granulomas, it is clearly evident that the predominant type of granulomas in the androgen treated groups is the cellular type [38% and 57.1%] in group III and IV respectively and this may reflect the possible beneficial effect of androgen on the diseased liver

Des-gamma carboxy prothrombin: a useful reliable early marker for detection of hepatocellular carcinoma

Hepatocellular carcinoma [HCC] is one of the most common cancers in the world. The early detection is important in the management of cancer. Des-gamma carboxy prothrombin also called protein induced by vitamin K absence or antagonist II [PIVKA II] has attracted attention as a marker for early diagnosis of HCC. to assess a diagnostic role of PIVKA II for early detection of HCC. The present case control study included 50 patients who fulfilled criteria for inclusion, with an age ranged from 30-60 years of both sexes. They were selected from outpatient clinic of the National Hepatology and Tropical Medicine Research institute [NHTMRI] from January 2003 to July 2004. Twenty age and sex matched subjects were used as controls. Patients were classified into 3 groups: Group I: 22 cases with HCC, Group EL: 15 late cases of decompansated liver cirrhosis, Group III: 13 early cases of liver cirrhosis. All patients were subjected to proper history taking, thorough clinical examination, and Laboratory investigations: complete blood picture [CBC], erythrocyte sedimentation rate [ESR], coagulation profile, liver function test and kidney function test. Determination of plasma PIVKA II, serum alpha fetoprotein [AFP] and serum ferritin were done by an ELISA technique. The collected data were computerized and analyzed statistically. The present study showed that level of- plasma PIVKA II was highly significantly increased in group I compared to the other groups [p < 0.01]. Group III showed a significant difference compared to control group [p < 0.05]. Serum AFP was highly significantly increased in group I compared with other groups [p < 0.01]. Also group II and group III showed a significant difference in serum AFP than control group. Serum ferritin was significantly higher in group I and group II than controls [p < 0.01]. Group II showed a significant increase in serum ferritin than group III [p < 0.05]. Plasma PIVKA II is a reliable marker of HCC that can be used with AFP as a complementry test for early detection of HCC in patients with cirrhosis