Antitrypanosomal potentials of the extract and fractions of Abrus precatorius seeds

ObjectiveTo evaluate the in vivo trypanocidal activity of the methanol extract and fractions of Abrus precatorius seeds in mice.MethodsParasiteamia was induced unto mice by intraperitoneal injection of 1.25×10⁵ Trypanosoma in normal saline. Five days when a high level of parasiteamia was established treatment commenced until ten days. The mice were treated with 10, 20 and 40 mg/kg bt. of the extract and 5 and 10 mg/kg bt. of the fraction (F₂), respectively for 5 days. Diminazene aceturate at the dose of 3.5 mg/kg bt. for two days was used as the reference drug. The level of parasitaemia and packed cell volume (PCV) of the animals estimated.ResultsAt doses of 10, 20 and 40 mg/kg the crude extract showed a sharp reduction in the level of parasitaemia in mice compared with the untreated group. The mice treated with F₂ at doses of 5 and 10 mg/kg showed a sharp reduction in the level of parasitamia to zero in day 9, and a gradual recovery from the 12th day of treatment. This effect is comparable to that of the mice treated with 7 mg/kg of standard drug diminazene aceturate. The PCV of the treated showed a gradual decrease with time, but not as much as the untreated group. Phytochemical screening revealed the presence of glycosides, alkaloids, carbohydrates, tannins and proteins in the Abrus precatorius powder while F₂ was rich in alkaloids.ConclusionsThis study shows that both the extract and the fractions of Abrus precatorius seeds exhibited a promising trypanocidal property. Alkaloids may be responsible for the observed activity.     

Retroviral transduction of human progenitor cells: use of granulocyte colony-stimulating factor plus stem cell factor to mobilize progenitor cells in vivo and stimulation by Flt3/Flk-2 ligand in vitro

The clinical application of gene transfer is hindered by the availability of the multipotential stem cells and the difficulty in obtaining efficient retroviral transduction. To assess potential means by which gene transfer into human hemopoietic stem cells might be enhanced, the retroviral transduction efficiency of human bone marrow cells (BM) or peripheral blood progenitor cells (PBPC) was compared at multiple time points after in vivo administration of granulocyte colony- stimulating factor (G-CSF). This was further compared with the transduction efficiency of cells mobilized with G-CSF plus stem cell factor (SCF) in a cohort of patients randomized to receive either one or two growth factors and with normal BM function. Using the LNL6 retrovirus, retroviral transduction efficiencies of up to 19% were observed for both PBPC and BM (n = 26 patients). There was at least a 100-fold increase in PBPC with G-CSF alone and a further 30-fold increase in the total number of progenitor cells available for retroviral transduction using the combination of SCF plus G-CSF. However, pretreatment of patients with G-CSF with or without SCF did not enhance the retroviral infectability of growth factor-mobilized progenitor cells. The effect of the growth factor, Flk-2/Flt3 ligand (FL), was also examined with respect to retroviral transduction efficiency of human progenitor cells. FL plus IL-3 in vitro increased the retroviral transduction efficiency up to eightfold compared with results observed using other combinations of cytokines tested (P < .001). These findings have clinical implications both for increasing the number of target cells for in vivo gene-marking/gene-therapy studies and improving the efficiency of gene transfer.     

Low-level hepatitis C viremia and humoral immune response to NS4 in chronic hepatitis B virus-hepatitis C virus coinfection

BACKGROUND: There is a limited amount of published data on the interference of hepatitis B virus (HBV) on hepatitis C virus (HCV). The aim of this study was to investigate the effect of concurrent HBV infection on serum titers of HCV RNA and HCV antibody profiles in chronic HCV infection. METHODS: The clinical and virological profiles (serum titers of HCV RNA, HCV genotypes and antibody profiles) of 25 patients with chronic HBV-HCV coinfection were compared with those of 25 age- and sex-matched patients with HCV infection alone. RESULTS: Among the 25 patients with HBV-HCV coinfection, only 3 were found hepatitis Be antigen (HBeAg) and HBV DNA positive by hybridization assays, and the other 11 were found HBV DNA positive by polymerase chain reaction. Genotype 1b was dominant in both HBV-HCV coinfection and HCV infection alone (64% versus 84%, P > 0.1). Patients with HBV-HCV coinfection had significantly lower alanine aminotransferase (ALAT) levels and inflammatory scores but higher fibrosis scores than those with HCV infection alone. Serum titers of HCV RNA were significantly lower in HBV-HCV coinfection than in HCV infection alone. The frequency and relative intensity of antibody response to core, E2/NS1, NS3, and NS5 showed no significant difference between the two groups, but antibody response to NS4 was diminished significantly in HBV-HCV coinfection. CONCLUSIONS: In HBV-HCV coinfection, serum levels of HBV DNA are usually low or undetectable. Concurrent HBV infection, however, could interfere with HCV replication and suppress antibody response to NS4. The biological significance of selective inhibition of humoral immune response to NS4 in HBV-HCV coinfection should be further studied.     

Comparison of different techniques of cataract surgery in bacterial contamination of the anterior chamber in diabetic and non-diabetic population

Aim:

To compare the bacterial contamination of the anterior chamber (AC) between manual small incision cataract surgery (SICS) and phacoemulsification (Phaco). To study the conjunctival flora and bacterial contamination of AC between well-controlled diabetics and non-diabetics.

Materials and Methods:

Three hundred and sixty-eight patients were randomized to manual SICS and Phaco. Sixty-eight patients were excluded for not completing follow-up or for intraoperative complications like posterior capsule rupture. One hundred and fifty patients in each group were finally analyzed. Conjunctival swabs were taken on admission, after one day of topical ofloxacin and 15 min after 5% Povidone Iodine (PI) instillation. AC aspirate at the end of the surgery was also cultured.

Results:

Fifty-six (18.66%) patients had positive conjunctival swab on admission which was reduced to 19 (6.33%) with topical ofloxacin and to five (1.66%) with instillation of 5% PI. AC contamination in both manual SICS and Phaco was 0.66%. The conjunctival flora in diabetics was similar to non-diabetics. None of the diabetics had AC contamination. Statistical analysis was performed by Chi-Square test (with Yates’ correction).

Conclusion:

Statistically significant reduction in conjunctival flora was achieved with topical ofloxacin and 5% PI instillation and AC contamination in both manual SICS and Phaco was minimal (0.66%). Well-controlled diabetics who underwent cataract surgery in this study had similar conjunctival flora and AC contamination as non-diabetics.     

Streptozotocin-Induced diabetes mellitus is associated with increased pancreatic tissue levels of noradrenaline and adrenaline in the rat

The pancreata of streptozotocin-induced diabetic rats were examined to determine whether the pancreatic tissue content of catecholamines is altered after the onset of diabetes. Experimental diabetes was induced by intraperitoneal injection of streptozotocin (60 mg/kg body weight). Four weeks after the induction of diabetes, pancreatic tissue fragments were taken from the tail end of the pancreas and processed for catecholamine content using the high-performance liquid chromatography method. Immunohistochemical analysis showed that the pancreata of diabetic rats contained more tyrosine hydroxylase-positive nerves compared with controls. Pancreatic noradrenaline content, expressed as the mean +/- SD, was significantly (p < 0.03) greater in diabetic rats (54+/-11.74 pg x mL(-1) x mg tissue(-1)) compared with normal, sex- and age-matched control rats (37.54+/-1.18 pg x mL(-1) x mg tissue(-1)). Similarly, the adrenaline content in diabetic rat pancreatic tissue (102.69+/-20.24 pg x mL(-1) mg tissue(-1)) was markedly greater (p < 0.003) compared with sex- and age-matched controls (35+/-9.23 pg x mL(-1) x mg tissue(-1)). In contrast, 5-hydroxyindole acetic acid decreased significantly (p < 0.0002) in diabetic pancreatic tissue (13.41+/-0.87 pg x mL(-1) x mg tissue(-1)) compared with controls (80.72+/-1.46 pg x mL(-1) x mg tissue(-1)). The plasma levels of these catecholamines also increased slightly but not significantly in diabetic rats compared with controls. These results suggest that diabetes is associated with increased noradrenaline and adrenaline and decreased 5-hydroxyindole acetic acid pancreatic tissue levels. These disturbances in catecholamine metabolism may play a role in the pathogenesis of the acute and chronic complications of diabetes mellitus.     

Is p53 gene mutation an indicatior of the biological behaviors of recurrence of hepatocellular carcinoma？

AIM: To evaluate mutant p53 gene in primary hepatocellular carcinoma and to investigate the correlation between it and the recurrence of hepatocellular carcinoma. METHODS: Mutations of p53 gene were examined using anti-human p53 monoclonal antibody and immunohistochemical staining in 79 resected hepatocellular carcinomas. The correlations among variables of p53 positivity and invasiveness, disease free interval and survival were studied. In addition, in those who developed recurrence, the correlation among p53 positivity, clinical features and post-recurrence survival were also studied. RESULTS: Of these 79 cases, 64 (81 %) had p53 mutation. Those patients with mutant p53 positivity had significantly more tumor recurrence (76.6 % vs 40.0 %, P=0.0107). However, the COX proportional hazards model showed that p53 overexpression had only weak correlations with recurrence free interval and survival time (P=0.088 and 0.081), which was probably related to the short duration of follow-up. The invasiveness variables may be predictors of HCC recurrence. On univariate analysis, more patients with mutant p53 positivity had vascular permeation (78.1 vs 40.0 %, P=0.0088, O.R. (odds ratio) =5.3), grade II-IV differentiation (98.4 vs 80.0 %, P=0.0203, O.R. =15.7), no complete capsule (82.8 vs 53.3 %, P=0.0346, O.R. =4.2) and daughter nodules (60.9 vs. 33.3 %, P=0.0527, O.R. =3.1) than patients with negative p53 staining. On multivariate analysis, only vascular permeation and grade of differentiation remained significant (P=0.042 and 0.012). There was no statistically significant correlation between the status of p53 in the primary lesion and the clinical features of recurrent hepatocellular carcinomas examined, including extrahepatic metastasis (P=0.1103) and the number of recurrent tumors (P=1.000) except for disease over more than one segment in the extent of recurrent tumors (P=0.0043). The post-recurrence median survival was lower in patients in whom p53 mutation had been detected in the primary lesion with a weak significance (3.42 months vs 11.0 months, P=0.051). CONCLUSION: Our findings suggest that p53 mutation correlates significantly with invasiveness including vascular permeation, grade of cellular differentiation, incomplete capsule and multinodular lesions. Hepatocellular carcinomas with p53 mutations had more tumor recurrence and p53 mutation may also influence disease recurrence interval and survival time. Hepatocellular carcinomas with p53 mutations recur more extensively with a shorter survival. Therefore, p53 mutation in the primary lesion is useful as an indicator of the biological behavior of recurrent hepatocellular carcinomas.     

Hepatitis C virus infection facilitates gallstone formation

BACKGROUND: Bile duct damage and hepatic steatosis are two characteristic histological findings in hepatitis C virus infection; and high prevalence of hepatitis C antibody is noted in patients with cholangiocarcinoma. The purpose of the present study was to examine the relationship between biliary diseases and hepatitis C virus infection. METHODS: Persons who received a general checkup in Chang Gung Memorial Hospital between 2000 and 2002 were included. All of them had hemogram, serum biochemistry, hepatitis B surface antigen, hepatitis C antibody and ultrasonography studies. The prevalence of gallbladder stone, bile duct stone and gallbladder polyp/cholesterolosis were compared in different viral infection groups. RESULTS: Of the 28 486 persons, 22 967 were negative for both hepatitis B surface antigen and hepatitis C antibody (group NBNC), 4152 were hepatitis B surface antigen carriers (broup B), 1195 were positive for hepatitis C antibody (group C), and 172 were positive for both markers. The 379 persons (1.3%) having had cholecystectomy were considered to have gallbladder stone at the time when cholecystectomy was done. Gallbladder stone was found in 6.0% persons of group NBNC, 5.4% in group B and 11.7% in group C. The prevalence of gallbladder stone in group C was found especially high for age groups 31-40 years and 61-70 years. The prevalence of bile duct stone was higher in group C (0.4%) than in group NBNC or B (both 0.1%). Stepwise logistic regression analysis showed that age, liver cirrhosis, body mass index, hepatitis C virus infection and gender were independent factors associated with gallbladder stone. CONCLUSIONS: Hepatitis C virus infection facilitates gallstone formation.