Immunofluorescent detection of apoptosis in chronic viral hepatitis and other liver diseases

Chronic viral hepatitis is a common health problem in many countries including Egypt. The microscopic appearance of chronic viral hepatitis is usually associated with the appearance of eosinophilic structures called apoptotic bodies. Apoptosis is a type of programmed cell death. Abnormalities of apoptosis might contribute to many disease processes, the most important of which is neoplasia. The aim of this work is to study the frequency of apoptosis in cases of chronic viral hepatitis C [HCV], B [HB], pure Bilharzial liver affection, combined Bilharzial and chronic viral hepatitis C in addition to cases of hepatocellular carcinoma [HCC]. Liver biopsies [10 cases of HCV+ve patients, 6 cases HBs Ag +ve patients, 8 cases pure Bilharzial liver affection, 10 cases of combined Bilharzial and HCV, 10 cases of HCC and 6 cases of normal liver tissue obtained from areas adjacent to focal hepatic lesions were used as control]. A modified method of the terminal deoxynucleotidyl transferase- mediated dUTP-biotin nick end labeling [TUNEL] using immunofuorescence [FITC - dUTP] was used to detect apoptosis. Apoptotic index was determined for each case by counting apoptotic cells per 1000 cells in different fields. Apoptotic index for control cases was 0.1 +/- 0.08, cases of HCV 0.23 +/- 0.125, cases of HB 0.25 +/- 0.1, pure Bilharzial cases 0.125 +/- 0.08, combined Bilharzial and HCV cases 0.24 +/- 0.126 and for cases of HCC, it was 0.08 +/- 0.07.The difference between apoptotic index in cirrhotic and non-cirrhotic cases was statistically non significant [P = 0.896]. As a conclusion, we found that the modified TUNEL method using fluorescein isocyanate labeled dUTP was useful and sensitive for detecting apoptosis in the liver. The apoptotic index increases significantly than the control in cases of chronic viral hepatitis C [P = 0.044], B [P = 0.024] and combined HCV and Bilharzial liver affection [P = 0.033]. Apoptosis in pure Bilharzial liver affection does not significantly differ from the control [P = 0.636]. In cases of HCC, apoptosis decreases than the control in a non-significant manner [P = 0.647]. Apoptotic index does not differ significantly between cirrhotic and non- cirrhotic cases

Does schistosomiasis affect cellular proliferation of hepatocytes in hepatitis C infection?

The role of schistosomal co-infection with HCV in affecting the course of chronic viral hepatitis is not clear. This study aims at evaluating the effects of schistosomal co-infection on histopathoiogical changes as well as cellular proliferation in liver biopsies of HCV infected patients. The study included 49 cases of chronic liver disease [14 cases of pure hepatitis C [HCV], 26 cases of combined HCV and shistosomiasis and 9 cases of pure schistosomal hepatic affection] .Histopathoiogical assessment of liver biopsies in cases of chronic viral hepatitis was done according to the Ishak's modification of Knodell's score for grading and staging system. Cellular proliferation was evaluated by immunohistochemical staining with the proliferation marker ki-67 [MIB-1] as well as by counting the mean number of argyrophilic nu-cleolar organizer regions [Ag NOR[s]]. Of the 14 cases of pure HCV, 9 cases [64.3%] showed minimal to mild chronic hepatitis and 5 cases [35.7%] showed moderate to severe chronic hepatitis.4 cases [28.6%] were cirrhotic. On the other hand,16 out of 26 cases of combined HCV and schistosomal infection [61.5%] showed minimal to mild chronic hepatitis while 10 cases [38.5%] showed moderate to severe chronic hepatitis. 8 cases [30.76%] were cirrhotic. These figures did not differ significantly from those of pure HCV infection with pvalues [0.236], [0.111] and [0.863] respecrively. As regards cellular proliferation, by using the immunohistochemical marker ki-67 [MIB-1], cases of pure HCV showed nuclear staining in 8/14 cases [57.14%]. The mean proliferation index PI was [5.14 +/- 5.66].In cases of combined HCV and schistosomal affection, 16 out 26 cases [61.5%] showed positive staining with PI [5.6 +/- 5.9]. The difference between the two groups was non significant [p=0.834]. In the group of pure schistosomal affection of the liver, 4/9 cases [44.4%] showed positive staining.The PI of this group was [3.11 +/- 4.25]. This figure does not differ significantly neither from the group of pure HCV [p = 0.254] nor from the group of combined HCV and schistosomiasis [p=0.195]. By using the traditional method for assessment of cellular proliferation by counting the mean number of Ag NOR[s], in the group of pure HCV, the PI was [1.62 +/- 0.13], that of combined HCV and schistosomiasis was [1.61 +/- 0.13] while that of pure schistosomal hepatic affection was [1.57 +/- 0.09]. The difference between the three groups was statistically not significant. This work shows that on the histopathological level, schistosomiasis does not significantly affect hepatic lesions induced by HCV. Moreover; shistosomiasis does not alter the proliferation index of hepatocytes of HCV infected patients. This work showed also that on assessing cellular proliferation, both the immunohistochemical method using ki-67 and counting the mean numbers of Ag NOR[s] give similar results

Epithelial cell proliferation in helicobacter pylori associated gastritis

Helicobacter pylori [H. pylori] is the main cause of chronic [type B] gastritis. The intestinal form of gastric cancer arises against a background of chronic gastritis. An increase in mucosal cell proliferation increases the risk of a neoplastic clone to emerge. However, little is known about cell proliferation in H. pylori associated chronic gastritis. The aim of this work is to evaluate cellular proliferation in cases of H. pylori associated chronic gastritis and to correlate the extent of cellular proliferation with different histopathological lesions induced by the organism. Antral endoscopic biopsy obtained from 47 patients with chronic gastritis having Ig G antibodies against H. pylori detected serologically by ELISA were collected. In them the presence of the organism was demonstrated by Hx and E as well as by Geimsa stain. At the same time 12 cases of seronegative subjects to H. pylori having histopathologicalty normal gastric mucosal biopsy were used as a control group. Cellular proliferation was assessed immunohistochemically using proliferating cell nuclear antigen [PCNA] and the obtained results were correlated to different histopathological changes of H. pylori associated chronic gastritis namely neutrophilic cellular infiltrate, round cell infiltration, presence of lymphoid follicles, gastric atrophy and intestinal metaplasia. Gastric mucosal epithelial cell proliferation increased significantly in H. pylori induced chronic gastritis than the control group [P<0.001].The proliferation index also increased significantly with increased intensity of round cell infiltration,[P=0.009], presence of mucosal lymphoid follicles [p=0.047] and degree of gastric atrophy [p=0.03]. Cell proliferation did not significantly change with the presence or the degree of neutrophilic infiltration. None of our cases revealed metaplastic change. The results of this study show that H. pylori induced chronic gastritis is associated with marked increase in gastric mucosal epithelial cell proliferation. The results of the study also suggest that in cases of H. pylori chronic gastritis, the density of round cell infiltration, the presence of lymphoid follicles and gastric atrophy are associated with increased cellular proliferation. These changes may play a part in gastric carcinogenesis

teratogenic study of selenium in albino rats

Teratogenecity of selenium was investigated in pups of 80 dams. Forty dams served as controls and the other forty administered orally and daily 1.5 mg/kg b. w. of selenium dioxide from day 6 to 15 of gestation. Uteri of sacrificed dams and pups of both groups were examined. Data obtained revealed increased number of resorped and dead pups per litter and decreased number of livings per litter. Significant increase [83.3%] of abnormal pups among livings was found. Growth retardation manifested by a 51.58% and 48.55% decrease in rate of weight and length gain per day recpectively. Also the size of litter decreased 19.8%. Selenium resulted in a 19.35% decrease in mean embryonic weight accompanied by a 16.9% decrease in crown-rump length. High percentages of morphological abnormalities were detected. They included small sized pups [83.3%]; wrinkled skin [74%]; kinking of tails [35.4%]; translucent skin [22.9%]; flat occiput [9.4%]; kyphosis [12.5%]; multiple haemorrhagic areas [67.7%]; misshaped heads [15.6%] and missing eye [9.4%]. Skeletal anomalies following prenatal administration of selenium included absent carpal and metacarpal bones [62.5%]; absent sternebrae [32.5%]; incomplete skeleton [47.5%]; incomplete ossification of cranium [47.5%]; absent coccygeal vertebrae [30%]; absent ischium [25%]; absent metatarsal bones [60%] and incomplete union of vertebral column [47.5%]. Histopathological abnormal changes were found in the skin and internal organs of pups of Se treated dams. Data of the present study proved the teratogenic potential of selenium in Albino rats

Studies on neurologic functional and structural changes in acrylamide toxicity [an experimental study]

This work was designed to study the toxic effects of acrylamide and glycidamide in vivo on the isolated Sprague Dawley rats to compare the toxic effects of acrylamide versus its epoxide metabolite, glycidamide on the peripheral nerves. Acrylamide toxicity was induced by a daily i.p. injection of 25 and 50 mg/kg body weight/day, respectively, for 8 days. The weight of the rats, performances of the rotarod test and hind limb splay was performed every other day. On the 9th day, the animals were sacrificed and sciatic nerves were removed and examined. The results are presented and described

Effects of chronic nicotine administration on male fertility in albino rats [an experimental study]

This work was designed to investigate the effects of intraperitoneal administration of nicotine in different doses over a period of 60 days on the fertility of adult male albino rats. Ninety-four adult male albino rats were used in this study and divided into four groups: A control group of ten rats received saline only and three test groups each containing 28 rats. Each of the test groups received a different dose of nicotine [0.5, 0.75 and 1.0 mg/kg b. w., respectively, every other day for 60 days]. Each group was further subdivided into three subgroups; the first [14 rats] was sacrificed immediately after 60 days, while the second and third subgroups [seven rats each] were sacrificed one and two weeks after stopping nicotine therapy. By the end of the experiment, blood samples were obtained from sacrificed rats and used for hormonal assay [testosterone and leutinizing hormone levels]. Then, the rats were subjected to epididymal spermatozoal examination and histopathological study of their testes. The results were presented and discussed

Effect of concurrent administration of acetaminophen with acetyl salicylic acid or indomethacin on the kidney, liver and stomach of rats

This study was conducted on 72 albino rats weighing averagely 100- 120 g. Theywere divided into 6 equal groups, the first group was served as control, the2nd, 3rd, 4th, 5th, 6th groups were given acetyl salicylic acid, indomethacin,acetaminophen, acetyl salicylic acid + acetaminophen and indomethacin +acetaminophen, respectively. Drugs were given orally in dose of 1/10 LD50daily for 30 days. After this period, the animals were subjected tobiochemical investigations [serum urea, creatinine, total bilirubin, ALT, AST]and histopathological studies [kidney, liver and stomach]. It was found thatconcurrent administration of acetaminophen with acetyl salicylic acid orindomethacin had good protection against gastric erosive effect of eitheracetyl salicylic acid or indomethacin, moreover, no exaggeration of thehepatic and renal impairment was detected