protective effect of insulin against isoflurane and sevoflurane induced hepatocellular apoptosis

Several studies deducted that inhalational anesthetics induce apoptosis in human cells. Insulin is believed to have an antiapoptotic action so it is widely used as cardioprotective agent against ischemic reperfusion injuries. This study compared the apoptotic effect of inhalational anesthetics and figuring out the antiapoptotic effect of insulin against perioperative induced hepatocellular apoptosis using immune histochemical study of liver biopsy. Eighty [ASA I] patients scheduled for laparoscopic cholecystectomy were randomly allocated into 4 groups [20 patients each]. Two groups were anesthetized using isoflurane and the other two were anesthetized using sevoflurane. The control groups [IC, SC] received normal saline and the insulin groups [II, SI] received glucose, insulin and potassium [GIK] infusion. Infusions were given 2 hours before induction of anesthesia. Liver biopsy was taken before the umbilical port closure. In liver biopsy Caspase 3, 7, 9 and Akt activity were evaluated. Liver function tests were estimated before infusion and one day after surgery. Serum K and blood glucose levels were closely monitored all through the study. The results showed that in the isoflurane groups, the percentage of caspase 3 and 7 positive cases decreased while Akt positive cases increased significantly in II compared to IC respectively [p < 0.05]. In the sevoflurane groups, the percentage of caspase 3 positive cases decreased significantly in SI compared to SC group [p < 0.05]

Infected freshwater snails, biomphalaria alexandrina, the intermediate host of schistosoma mansoni

The objective was to study the chromosomes of Biomphalaria alexandrina snails [class Gastropoda] in control and infected state which could be helpful in understanding how host-parasite relationships in feasible and effective control measures. The chromosomal changes were studied using the air-drying method. The results showed that B. alexandrina had a diploid chromosome number, 2n = 36. Also, the meiotic stages were detected as early- leptotene and late leptotene, zygotene, diplotene, metaphase I. The result also revealed the presence of a primitive sperm, with a conical head and a very long, uni-flagellate tail. A comparative meiotic chromosome analysis between the control and infected ones showed some significant differences, as pachytene and diplotene were more condensed

Ameliorative Effect of Bone Marrow-Derived Stem Cells on Injured Liver of Mice Infected with Schistosoma mansoni

The technique of stem cells or hepatocytes transplantation has recently improved in order to bridge the time before whole-organ liver transplantation. In the present study, unfractionated bone marrow stem cells (BMSCs) were harvested from the tibial and femoral marrow compartments of male mice, which were cultured in Dulbecco's modified Eagle's medium (DMEM) with and without hepatocyte growth factor (HGF), and then transplanted into Schistosoma mansoni-infected female mice on their 8th week post-infection. Mice were sacrificed monthly until the third month of bone marrow transplantation, serum was collected, and albumin concentration, ALT, AST, and alkaline phosphatase (ALP) activities were assayed. On the other hand, immunohistopathological and immunohistochemical changes of granuloma size and number, collagen content, and cells expressing OV-6 were detected for identification of liver fibrosis. BMSCs were shown to differentiate into hepatocyte-like cells. Serum ALT, AST, and ALP were markedly reduced in the group of mice treated with BMSCs than in the untreated control group. Also, granuloma showed a marked decrease in size and number as compared to the BMSCs untreated group. Collagen content showed marked decrease after the third month of treatment with BMSCs. On the other hand, the expression of OV-6 increased detecting the presence of newly formed hepatocytes after BMSCs treatment. BMSCs with or without HGF infusion significantly enhanced hepatic regeneration in S. mansoni-induced fibrotic liver model and have pathologic and immunohistopathologic therapeutic effects. Also, this new therapeutic trend could generate new hepatocytes to improve the overall liver functions.

Expression of matrix metalloproteinase 1[MMP1] in hepatocellular carcinoma [HCC]: Immunohistochemical and Biochemical Studies

Background: The present study aimed to evaluate the expression of Matrix Metalloproteinase-1 [MMP1] in HepatoCellular Carcinoma [HCC] by using the Immunohistochemical technique, which allows us to integrate the biological aspects of this enzymatic expression in the morphological context of HCCs

Material and Methods: The study was performed on 70 subjects from out and in patients of Tropical medicine Department, Thiodor Billhars Institute during the period from January 2011 until June 2012. The present study included 60 patients with chronic hepatitis C who had undergone liver biopsy. They consisted of 42 men and 28 women with ages ranging from 36 to 66 years. The diagnosis of chronic hepatitis C was made on the basis of positivity for anti-HCV [by the second generation ELISA], and confirmed by HCV-RNA reverse transcription polymerase chain reaction [RT-PCR]. Patients were divided into four groups: Group I: included 10 normal persons with no history of liver disease with normal liver enzymes and free ultrasonographic finding as normal control. It included 6 males, 4 females, with ages ranging from 34 to 48 years. Group II: included 20 HCV infected patients without cirrhotic changes. It included 11 males, 9 females, with ages ranging from 39 to 53 years. Group III: included 20 HCV infected patients with liver cirrhosis, 12 males, 8 females, with ages ranging from 48-63 years. Group IV: included 20 HCV infected patients with HCC, 16 males, 4 females, with ages ranging from 53-64 years

Results: Blood Picture, [Hb, WBCs, RBCs, Plts, PC and ESR]. Liver Function Test [ALT, AST, ALB, GGT, ALP, T. BIL and D. BIL]. Matrix Metalloprotenase 1[MMP1] Measurements: Serum MMP1. Histopathological investigation Including histopathological changes in the liver tissue

Conclusion: our results suggest that MMP-1 is overexpressed in a large proportion of patients with HCC and the high expression level of protein correlated with the disease progression and poor clinical outcome in HCC. Furthermore, MMP-1 high expression proved to be a risk factor for tumor recurrence and independent molecular marker of prognosis in HCC and may become a novel target in the strategies for the prediction of tumor progression and prognosis of this disease

Evaluation of the diagnostic value of serum and tissue apoptotoic cytokeratin- 18 in patients with chronic hepatitis C

Hepatitis C virus [HCV] is considered the most common aetiology of chronic liver disease [CLD] in Egypt. The disease severity ranges from mild illness to cirrhosis and hepatocellular carcinoma. A role for apoptosis in liver damage caused by HCV chronic infection has been suggested. Cytokeratin 18 [CK-18] is the major intermediate filament protein in the liver and is a known caspase substrate in hepatocyte apoptosis. Therefore, we analysed the serum and tissue levels of CK-18 in patients with chronic HCV infection to evaluate its role in hepatocyte apoptosis. We also correlated CK-18 expression with the severity of hepatic pathology. This study examined 80 Egyptian patients with liver disease. There were 69 patients with chronic hepatitis C and 11 patients with hepatitis C-induced cirrhotic changes. Fifteen healthy controls were also included in the study. The levels of CK-18 fragment were quantified in paired serum and liver biopsy samples. The serum and tissue CK-18 levels were reduced in chronic HCV patients compared to early cirrhosis patients. This result indicates that serum levels of CK-18 and the hepatic expression of CK-18 might play an important role in disease progression. The serum and tissue levels of CK-18 were significantly increased and directly correlated with inflammation severity, stage of fibrosis, and ALT levels in the chronic HCV group and the cirrhotic liver group. There was no significant difference in viral load between patient cohorts. The serum level and the hepatic expression of CK-18 are related to disease activity and are directly correlated with METAVIR scoring. This result suggests that serum CK-18 levels may be useful for monitoring disease activity in chronic HCV and liver cirrhosis patients

Early detection of hepatocellular carcinoma on top of liver cirrhosis: the fas receptor and ligand system

Hepatocyte aberrations, accumulation of chromosomal damage and possibly initiation of hepatic carcinogenesis is thought to be caused by the continued viral replication and the persistent attempt by a less than optimal immune response to eliminate hepatitis C virus [HCV] infected cells. The identification of the "death factors" including Fas and its Ligand [Fas-L] as a major regulator of both apoptosis and immune function has provided insight into an attractive mechanism of tumor escape from immune clearance. To assess the hepatic expression of Fas/Fas-L, the Fas receptor [Fas-R] expression on lymphocyte, and serum soluble Fas [sFas] in an attempt to analyze the role of Fas receptor/ligand system in the multistep process of fibrosis/carcinogenesis and the possible use of the serum marker as possible candidate biomarkers for an early detection of hepatocellular carcinoma [HCC]. The current study included 100 samples from cases at Theodor Bilharz Research Institute and Kasr Al Aini Hospital in Egypt. There were 90 cases of chronic hepatitis C [CHC] infection [and negative hepatitis B virus infection]. There were 30 cases without liver cirrhosis, 30 cases with liver cirrhosis and 30 cases with HCC. 10 liver biopsies were taken from healthy livers as normal controls. Histopathologic study and immunohistochemistry for detection of hepatic Fas and Fas-L expression were determined for all cases. Electron microscopy [EM] and immunoelectron microscopy [IEM] examination for detection of Fas-R expression on lymphocytes were also done. sFas, liver function tests, serologic markers for viral hepatitis, and serum alpha-fetoprotein level [alpha-FP] were done. The sFas in both HCC and CHC with cirrhosis patients were significantly higher than those of normal controls and CHC without cirrhosis [P<0.01], but there was no significant difference between the cirrhosis and HCC patients. Positive hepatic expression of both Fas and Fas-L were significantly increased in the diseased groups [p<0. 01] compared to the control specimens. A progressive Fas and FasL increase from CHC without cirrhosis to CHC with cirrhosis followed by a decline from the latter to HCC. Apoptotic Fas and Fas-L proteins expression was significantly increased with the necroinflammatory activity and the advancement of fibrosis. There was a non-significant negative correlation between sFas and hepatic Fas. In addition a significant over expression of Fas-R on separated lymphocytes was associated with a higher frequency of apoptotic cell death as detected by EM examination. Conclusion: The Fas receptor/ligand system was significantly involved in the process of liver cirrhosis converting into HCC. Down-regulation of Fas expression, up-regulation of Fas-L expression in hepatocytes and elevation of serum sFas level were important in tumor evasion from immune surveillance and in hepatic carcinogenesis

Ultrastructural study of human umbilical cord blood Buffy coat and Wharton's Jelly cells

Human umbilical cord [UC] has been a tissue of increasing interest in recent years. Umbilical cord blood [UCB] was used in the treatment of several hematological diseases. New potential uses of cord blood in nonhematopoietic applications are now proposed. Lately, the Wharton's jelly [WJ], the embryonic mucuos connective tissue of the UC, with its mesenchymal stromal cells represents a promising source of cells for several regenerative therapies. This study aims to highlight the morphological ultrastustural characteristics of the UCB buffy coat and the WJ stromal cells in situ. Tiny specimens from the UCB buffy coat and the WJ were double-fixed with glutaraldehyde and osmium tetroxide, dehydrated, and embedded in epoxy resin. Ultrathin sections were cut and stained with uranyl acetate and lead citrate. Transmission electron microscopic examination [TEM] of the UCB buffy coat cells showed immature cellular constituents and few changes in the ultrastructural morphology of cord blood platelets. These ultrastructural changes may clarify the specific cultural, immunological and regenerative properties attributed to cord blood in comparison to bone marrow or peripheral blood. Wharton's Jelly TEM showed stromal cells exhibiting morphological resemblance with both fibroblasts and smooth muscle cells. In addition, cells resembling stellate cells exhibiting intracytoplasmic lipid globules and ultrastructural properties related to protein synthesis were discerned. Also, oval cells with primitive nuclear and cytoplasmic features pointing to a subpopulation with stem cell potency were seen. We suggest that the WJ contains several distinct populations of stromal cells with different potentials including a primitive subpopulation. We propose that the nomenclature of these cells should not only be linked to their morphological appearance but also to their specific functional properties. Extensive in situ and in vitro molecular, immunohistochemical and immuno electron microscopic studies are necessary to clearly identify these cell populations with different potentials

Angiogenic potential of human umbilical cord blood ex vivo expanded CD 133* stem cells in chronic hepatic fibrosis of murine schistosomiasis

As a source of hematopoietic stem cells [HSCs], umbilical cord blood [UCB] has the advantages of speed of availability, tolerance of more than one HLA mismatch, and a low incidence of severe graft-versus-host disease [GVHD]. Hence, it represents a promising, alternative non-costly and non-invasive source for prospective stem cell based therapy. In this study we investigated the angiogenic potential of ex vivo expanded human umbilical cord blood CD 133* stem cells transplanted into mice with chronic hepatic fibrosis induced by Schistosomiasis infection. Histopathological, ultrastructural and immunohistochemical analysis of mice liver sections were done to detect specific human angiogenic markers. Umbilical cord blood was obtained from healthy pregnant females after delivery and mononuclear cells were collected by density gradient using Ficoll Hypaque. Enrichment for the CD 133* stem cells was done by positive selection using the Magnetic Activated Cell Sorting system and magnetic microbeads. Cells were cultured in prirnaly ex vivo expansion medium for three weeks. Flowcytomeric analysis of the cultured cells was done in each step to identify the CD 133* cells. Schistosomiasis was induced in Swiss Albino mice by intradermal injection of schistosoma cercariae. Twenty two weeks post schistosoma infection a total of 0.3 x 106 human CD 133* stem cells were injected intrahepatically in mice. Accordingly, mice were divided into three groups: Group 1 [infected, transplanted]; Group 2 [infected controls] and Group 3 [healthy, transplanted]. All mice were sacrificed 3 wks after cell transplantation was done in groups I and 3. Histopathology and Electron microscopy showed an obvious increase in the capillary network and the small blood vessels and a decrease in the fibrosis known for this stage of the disease. By immunohistochemical analysis the cellular constituents of these newly formed blood vessels showed positive expression of the human-specific angiogenic markers CD31, CD34 and von Willebrand Factor [vWF]. Few hepatocyte like polygonal cells showed positive expression of human Vascular Endothelial Growth Factor [VEGF] and inducible Nitric Oxide Synthase [iNOS]. Ex vivo expanded CD 133* human stem cells incorporate into the liver of schistosoma infected mice enhancing local angiogenesis and hepatic neovascularization. These preliminary results obtained suggest a dual benefit of CD 133* cells in cell therapy in hepatic diseases based on its capability of hematopoietic and endothelial differentiation. We suggest that the CD 133* cells contribute to repair in a paracrine manner by creating a permissive environment that enables rapid proliferation and survival of damaged cells rather than through direct differentiation to hepatocytes

Gastrointestinal stromal tumors [GISTS]: clinical presentation, diagnosis, surgical treatment and its outcome

This study included 13 selected patients treated by surgical excision for lesions that proved postoperatively to be gastrointestinal stromal tumors [GISTs] by histopathological and immunohistochemistry studies. The demographic, clinical and operative reports data were collected. Eight cases were gastric GISTs, four cases were small bowel GISTs [jejunum 1 and ileum, 3] and GIST of the sigmoid colon was in one patient. Eight cases presented at the emergency department due to hematemesis [3], gastrointestinal obstruction [3], bowel perforation [1] and severe bleeding per rectum [1]. Three cases presented with a feeling of abdominal fullness and ill-defined palpable abdominal mass. Two cases were discovered incidentally during GIT endoscopy for dyspepsia. Diagnosis of GISTs was presumed on clinical basis and operative findings from gross morphological features. Complete resection [RO] was achieved for 12 tumors [92.3%]. The immunohistochemistry profile was positive for C-kit for all cases. One operative death was due to massive pulmonary embolism. Postoperative complications occurred in three [23%] as upper GIT bleeding [1], biliary gastritis [1] and wound infection [1], and one [7.69%] of ileum tumor recurrence

role of probiotics in controlling giardia intestinalis in experimental animals

Efficacy of Lactobacillus acidophilus [Lactospore] for the control Research Institute Warak of Giardia intestianlis infection was evaluated in hamsters. Each El-Hadar Imbaba hamster was infected orally by 10.000 Giardia lamblia cysts. Animal were divided into five groups: Group A: control infected, untreate group. Group B: infected receiving Metronidazole. Group C: infected and receiving Lactospore. Group D: infected and receiving combination of Metronidazole and Lactospore. Group E: receiving Lactospore 7 days before infection acting as a prophylactic group. Groups B, C and D were given the appmpriate drug, three weeks post infection. Two weeks later stool analysis was performed and cysts/gm stool were counted, after which scarification of all groups took place. There was a highly significant difference between control and all treated groups. The highest percentage of reduction [cure rate] was in group D [98.6%]. followed by group B which gave a reduction rate of [93.8 1%]. The effect of the drugs on the vegetative [trophozoite] forms in the small intestine of sacrificed hamsters was studied. Combination of both drugs [group D] revealed a high significant cure rate [99.32%]. On the other hand when metronidazole and lactospore were given alone a reduction rate of 92.22% and 63.4% respectively was observed. The prophylaxis effect of Lactospore was highly noticed with 93.65% parasite reduction. Assessment of cure was also performed by electron microscopic and histopathological examination of the small intestine, peyer's patches and the spleen. Ultrastructural examination of the small intestine revealed remarkable destruction of the intestinal cell projection by Gardia cyst. Partial healing of the destructeci intestinal cell projection by Metronidazole was obvious, while complete healing could be detected in both groups D and E, Combined treatment with metronidazole and lactospore revealed remarkable activation of lymphocytes and macrophages. This easy flow through the endothelial cells lining the sinus in its way to the lumen to enhance their opportunity to overcome the infection could be detected. Histopathological examination revealed complete healing of the intestinal mucosa after the combined treatment while partial healing of the lining epithelium of the intestine was noticed after metronidazole treatment. The present study proves the efficacy of lactospore as a prophylactic agent for Giardiasis when given 7 days pre infection. This might be of considerable interest in eases of travelers diarrhea. In addition, it can strengthen the effect of metronidazole when both drugs are given together

Epidemiologic approach for early detection and control of renal and urinary tract diseases in rural populations

The dipstick testing, microscopic examination of urine and urine cytology were performed for inhabitants from two rural villages [El Shobak El Sharki, V.I and El Katta, V.2] in Giza G. The proliferating cell nuclear antigen [PCNA] and Schistosoma haematobium antigen were done by immuno-histochemical stain to confirm diagnosis. Also, they were subjected to medical questionnaire, clinical examination, ultra-sonography of kidneys and urinary tract. The results showed that V.2 had higher percentage of haematuria, proteinuria, glucosuria and lower urinary tract infection than V.I. Crystaluria was higher in V.I. Sensitivity of dipstick testing compared to microscopic examination was 26.6%, and specificity was 78.7%. Lower urinary tract infection cytologically detected was 44.2% sensitivity and 62.5% specificity compared to pyuria detected by microscopic examination of urine. Among those suffering variable urinary abnormalities, schistosome antigen was not detected in any fixed urine samples in comparison to corresponding confirmed positive controls. Urine cytology detected urinary tract infection, Crystaluria, dysplasia and atypia, squamous metaplasia and transitional cell carcinoma [TCC]. PCNA positivity was found in TCC [100%], dysplasia [50%] and squamous metaplasia [28.6%]. So, microscopic examination of urine proved valuable for tract abnormalities as pyuria, haematuria and crystaluria. Also, urine cytology is a must for malignancy of urinary tract especially in adult males

Estimation of cyclo-oxygenase-II protein in oesophageal mucosa in cases of gastro-oesophageal reflux disease, compared with normal oesophageal mucosa

Cyclo-oxygenase II [COX II] is an inducible enzyme. Its expression is increased during states of inflammation or carcinogenesis. THE AIM of this study was to demonstrate the expression of COX-Il protein in cases of reflux oesophagitis and compare it with normal oesophageal mucosa, Thirty two patients with GORD, and ten controls had an upper gastrointestinal endosopy. Mucosal biopsies were taken from lower oesophagus, and examined histologically for evidence and grading of GORD. COX II expression was evaluated using immunohistochemical staining of the mucosal biopsies. The results Showed a highly significant statistical difference between patients and control subjects in both endoscopic and histopathological findings [P < 0.01], and there was a positive correlation between both procedures [P < 0.001]. There was also highly significant statistical difference between patients and controls as regards all parameters of COX-II expression [P < 0.01]. COX II excpression was positively correlated with the degree of oesophagitis according to the degree of histopathological inflammation [P < 0.001] History of smoking was also associated with higher expression of COX II in all subjects examined [P < 0.01].The of this study showed that there is increased expression of COX II enzyme protein in oesophageal mucosa in cases of oesophagitis and that this might suggest the possibility of a causal relation. It also showed the increased expression in smokers, whether this proves to be of significance still awaits further studies

pharmacological approach to reverse portal hypertention and hepatic schistosomal fibrosis in Egypt, control experimental study

Schistosoma mansoni is the most prevalent cause of liver fibrosis in Egypt. It is characterized by hepatocyte damage, inflammation and chronic parasite egg-induced granuloma formation leading to fibrosis. Its management, particularly fibrosis, has focused primarily on treating and preventing the complications of portal hypertension. Unfortunately, there is no therapy that has been proved to prevent progressive hepatic fibrosis which is associated with a significant morbidity and mortality due to granulomatous hypersensitivity to parasite eggs. However, recent developments in understanding hepatic fibrogenesis confirm that recovery from advanced fibrosis is possible. There is a considerable imperative to develop anti-fibrotic strategies that are applicable to liver fibrosis. It was noted that a marked increase in the amount of different interstitial collagens types are associated with the development of fibrotic liver diseases. Mean while, it has been suggested that as long as the relative portions of liver collagen are still within the normal limits, the fibrosis may still be reversible. If it exceeds the normal limits fibrogenesis will proceed to its end stage, even if the etiological agent is removed. Collagen type IV and procollagen type III are two of the most accurate fibrosis markers which allow reliable non-invasive diagnosis. The T lymphocytes and the immuno-regulatory cytokines may be important in the host response to S. mansoni granuloma formation and fibrosis. Chronic parasite egg-induced granuloma formation can lead to fibrosis, which is immunologically characterized by the dominant Th2 response. Corticosteroids and prostaglandins interfere with both efferent and afferent mechanisms of immune function. These data indicate that this adjuvant therapy can be a candidate for therapeutic intervention in hepatic fibrosis through induction of a balance between Th1 and Th2 cells response as will be documented by the fibrosis markers One hundred S. mansoni infected hamsters [150-250 gm] were obtained from the BRPU-TBRI [5 groups, 20 hamsters each]. Treatment was started 10 weeks post infection. First G [20 hamsters] was neither infected nor treated, second G. was infected but untreated, third group infected and PZQ treated, fourth G. infected and PZQ and MP treated and fifth group infected and PZQ and PgE1 treated. Samples [liver and blood] were obtained 20 weeks post infection. The serum level of: liver functions, procollagen type III, collagen type IV and Th1 cytokine [IL-2] and Th2 cytokine [IL-b] were performed. Histopathology was performed to study live fibrosis, measuring the proliferate activity of the hepatocytes using cell image analyzer system and granuloma cells using the indirect immuno-histochemistry by monoclonal antibody proliferating cell nuclear antigen [PCNA]. In this study, G. V showed high significant reduction in granuloma size, type and percentage of fibrosis and significant elevation in percentage of degenerated ova compared to Gs. III and IV. The proliferation index measured using PCNA showed high proliferative activity of hepatocytes in non treated group which declined in the treated Gs. III, IV and V. The proliferation activity of hepatocytes and granuloma forming cells decreased significantly in G.V compared to G.IV. There was a significant reduction in liver function tests even tendency for normalization in G.V compared to group III and IV. Procollagen type III and collagen type IV were significantly low in the serum in G.V compared to Gs. III and IV. Th1 [IL-2] level was significantly high in G.V compared to Gs. III, IV and Th2 [IL-10] was significantly low in G.V compared to Gs III and IV indicating the low amount of fibrosis was in the group treated with PZQ and PgE1.PgE1 with PZQ to treat S. mansoni infected hamsters can modulate liver fibrosis and improves the liver function tests up to normalization. The balance between Th1 and Th2 cytokines level could be modulated to help reverse or decrease fibrosis in S. mansoni infected hamsters. This may pave the way for clinical application as combined therapy PZQ and PgE1 may by an effective approach to reverse hepatic fibrosis in schistosomiasis by the induction of dominant Th1 response