Making and using inexpensive manually constructed tissue micro-array: Experience of a tertiary care hospital in India

Background: Tissue micro-array enables the analysis of a large number of tissues simultaneously. Widespread use of this technology is hampered by the high cost of commercial array instruments. We describe our experience of constructing tissue micro-array in a simple method using easily available and inexpensive instruments. Materials and Methods: We used an 11-19 gauge (G) bone marrow trephine biopsy needle/ small sized slotted screwdriver to punch holes in the wax blocks. Cores were taken from donor tissue blocks using a bone marrow trephine biopsy needle and arrayed into host paraffin wax blocks. A detailed database was constructed for each array constructed. Results: The array blocks were used over a period of one year as internal control for immunohistochemistry (IHC), quality control and research. It took about 10 minutes to construct a nine-dot array and about one hour for a 56-dot array. During IHC, the average loss of control dots was less than one per cent. We did not see any loss of antigenicity in the control sections even after four weeks storage. Discussion: Tissue array construction by the technique described here is inexpensive and reliable alternative to automated instruments. Because it is easy to modify the arrays by varying the core size, it is easy to adapt this to individual labs and requirements. We recommend using blocks with cores in 3 x 3 to 5 x 4 grids as controls in IHC and for standardizing antibodies and array blocks with a larger number of cores for research

Adenomyoma of common bile duct arising in a type I choledochal cyst

Adenomyoma can be misdiagnosed as an adenocarcinoma, leading to needless and extensive surgical resections. A 45-year-old woman presented with right hypochondrial pain. Magnetic resonance imaging showed a choledochal cyst. Excision of choledochal cyst with Roux-en-Y hepaticojejunostomy was performed. A segment of dilated common bile duct and an attached nodule was received. Sections from the choledochal cyst showed a cyst wall composed of dense fibrous tissue lined by partially ulcerated columnar epithelium. Sections from the nodule showed interlacing whorls of smooth muscle bundles with entrapped glands. The glands were lined by cuboidal to columnar cells without nuclear atypia. This was recognized as an adenomyoma. To the best of our knowledge, this is the first reported case in which an adenomyoma was found associated with a type 1 choledochal cyst. A review of the existing literature and discussion of theories of genesis and the diagnostic pitfalls are presented

Comparison of clinical, biochemical and histological features of alcoholic steatohepatitis and non-alcoholic steatohepatitis in Asian Indian patients

<b>Background:</b> Alcoholic steatohepatitis (ASH) and non-alcoholic steatohepatitis (NASH) are significant forms of liver disease and may progress to end-stage liver disease, cirrhosis and potentially malignant complications. The most difficult aspect of establishing a diagnosis of NASH is distinguishing it from ASH. Laboratory markers such as AST, ALT and GGT lack sufficient sensitivity and specificity. <b>Aim:</b> To study the clinical, biochemical and histological differences between non-alcoholic steatohepatitis (NASH) and alcoholic steatohepatitis (ASH). <b>Materials and Methods:</b> Sixty histologically confirmed cases of non-alcoholic steatohepatitis and 38 cases of alcoholic steatohepatitis were included in the study. A modified form of scoring system proposed by Yip and Burt was used to grade histological features of NASH and ASH. <b>Results:</b> Mean age was 42.85 &#177; 12.36 years in ASH group and 35.07 &#177; 8.06 years for NASH group. Male: Female ratio was 37:1 in ASH and 4:1 in NASH. The mean ALT (<i>P</i> = 0.012), SAP (<i>P</i> = 0.003), serum bilirubin (<i>P</i> = 0.001), AST/ALT ratio (<i>P</i> = 0.03), steatosis (<i>P</i> &lt; 0.001), ballooning degeneration of hepatocytes (<i>P</i> &lt; 0.001), portal inflammation (<i>P</i> &lt; 0.001), Mallory hyaline (<i>P</i> = 0.001), ductular proliferation and fibrosis (<i>P</i> &lt; 0.001) showed a significant difference between ASH and NASH cases. <b>Discussion:</b> Older age, male sex, larger derangement of serum biochemistry, high serum bilirubin, AST/ALT &gt; 1, more ballooning degeneration, portal inflammation, Mallory&#x2032;s hyaline, hepatocytic and ductular cholestasis, ductular proliferation and higher stage of fibrosis favors a diagnosis of ASH. Younger age, high ALT, AST/ALT &lt; 1, higher grade of steatosis and absence of extensive neutrophilic portal inflammation favors a diagnosis of NASH

Independent predictors of steatohepatitis and fibrosis in Asian Indian patients with non-alcoholic steatohepatitis

Aim: To identify the clinical and biochemical predictors of histologic disease severity in patients with non-alcoholic steatohepatitis (NASH). Method: Clinical, anthropometric and biochemical data of 71 consecutive patients with a histological diagnosis of NASH were retrieved from the medical database. The histologic criteria proposed at the American Association for the Study of Liver Diseases Conference (2003) were followed. Histologic features and the necro-inflammatory grade of liver biopsy were scored in accordance with the Brunt score. Statistical analysis was performed to identify predictors of disease severity. Results: The male to female ratio of the study cohort was 3.1:1. The mean body mass index (BMI) of the patients was 25.67±1.9 kg/m2. In terms of necro-inflammatory grading, 21 (29.6%) patients were classified as grade 1, 26 (36.6%) as grade 2 and 24 (33.8%) as grade 3. Multivariate analysis revealed that BMI (P = 0.009), waist circumference (P = 0.035), waist:hip ratio (P = 0.011) and aspartate aminotransferase levels (P &lt; 0.001) were independent predictors of necroinflammatory grade and that female gender (P = 0.02), serum alkaline phosphatase levels (P = 0.018), cholesterol levels (P = 0.048) and low-density lipoprotein (LDL) levels (P = 0.025) were independent predictors of fibrosis stage. Conclusion: Female gender, BMI, waist:hip ratio, hypercholesterolemia and LDL levels are independent predictors of disease severity in patients with NASH and may influence the decision to biopsy

Oral leiomyoma

Oral leiomyoma is a benign smooth muscle tumor with a low incidence. Oral leiomyomas present as slow growing, asymptomatic sub mucosal masses, usually in the tongue, hard palate or buccal mucosa. They may be seen at any age and are usually discovered when they are 1 to 2 cm in diameter. The diagnosis is mainly determined by histological studies and special stains that confirm the smooth muscle origin. Surgical excision appears to be the best line of treatment and recurrence is unexpected. The purpose of this article is to present a case of a 32-year old male with a 1 month history of a leiomyoma on his right submandibular region

Evaluation of adefovir & lamivudine in chronic hepatitis B: Correlation with HBV viral kinetic, hepatic-necro inflammation & fibrosis

Background &amp; objectives : Chronic hepatitis B is an important cause of morbidity and mortality. We conducted a study comparing the efficacy of adefovir and lamivudine with respect to their impact on serum and hepatic viral DNA clearance, and improvement in hepatic necro-inflammatory score, in naive patients of chronic hepatitis B. Methods: This prospective randomized pilot study was conducted in Lok Nayak Hospital, New Delhi, involving 30 patients of chronic hepatitis B (both e antigen positive and negative); 15 were randomly selected to receive either adefovir or lamivudine for a period of 6 months. Quantification of serum and hepatic HBV DNA levels was done by real time PCR and liver biopsy was done at the beginning and end of 6 months. Results: Serum ALT was elevated to 2 or more times normalized in both the groups. In the adefovir group, two patients became HBeAg negative. In the lamivudine group, one patient became HBeAg negative. After therapy HBV DNA was negative in 26.7 per cent patients from adefovir group and 13.3 per cent patients from lamivudine group. Serum HBV DNA levels were correlated with the hepatic levels before therapy (r=0.843; P&#60;0.001) and after therapy (r=0.713, P&#60;0.001) showing strong correlation. There was a median reduction of 1.92 and 2.06 log copies per ml in serum HBV DNA load after adefovir and lamivudine therapy, respectively. The mean reduction in the histotogy activity index (HAI) score was 2 and 1.53, fibrosis score was 2.33 and 3.06 after adefovir and lamivudine therapy respectively. Interpretation &amp; conclusions : Adefovir and lamivudine treatment caused biochemical and serological improvement when administered for about 6 months with significant reduction in HBV DNA, serum and hepatic viral load without completely clearing the virus from either serum or liver. It also helped in reduction of the necro-inflammatory and fibrosis score of patients with chronic hepatitis B. Our study also showed significant correlation between serum and hepatic HBV DNA levels both before and after therapy. There was not enough evidence to show therapeutic advantage of one drug over the other in any of the parameters measured

Steatosis in chronic hepatitis B: Prevalence and correlation with biochemical, histologic, viral, and metabolic parameters

Background and Aims: Hepatic steatosis (HS) is highly prevalent in chronic hepatitis C and is an important variable predicting progression of histological injury, insulin resistance, and reduced response to antiviral therapy. There are limited data on HS in patients with chronic hepatitis B (CHB). This is relevant since response to current antiviral therapies for CHB is rather limited. We investigated the spectrum and predictors of HS in CHB patients. Materials and Methods: Liver biopsies of consecutive patients of chronic Hepatitis B Virus (HBV) infection were studied and were categorized as: Group I - hepatosteatosis (>5%) and Group II - no steatosis (£5%). Anthropometric, histological, biochemical, virological, and metabolic determinants were compared. Logistic regression analysis was applied to identify variables that were independently associated with the presence of steatosis. Results: Of the 350 patients, 118 (33.7%) liver biopsies showed steatosis (Group I); 65 (55.1%) had mild (6 to <25%) and 53 (44.9%) had moderate to severe steatosis (325%). Patients in group I, compared with group II, were older (35.5 ± 10.5 vs 27.9 ± 14.0 years, P < 0.01), predominantly male (M: F, 10.8: 1 vs 4.8: 1, P = 0.035), obese (75.0% vs 23.4%, P < 0.01), with higher body mass index (25.2 ± 4.8 vs 20.4 ± 3.5, P < 0.01), with higher triglycerides (138.8 ± 62.1 vs 88.0 ± 27.9, P = 0.02), with higher cholesterol (171.9 ± 43.5 vs 139.3 ± 37.6, P = 0.017), and with higher serum insulin (13.1 ± 9.1 vs 9.1 ± 6.0, P < .027) levels. HBV DNA level was significantly lower in group I than group II; however, HBV genotype did not influence HS. By multivariate regression analysis, only high serum triglyceride level was independent parameter associated with HS. Conclusions: Steatosis is seen in one-third cases with HBV-related chronic liver disease and is associated with host metabolic factors, especially serum triglyceride levels, whereas HBV DNA level negatively correlated with HS

High viral load and deregulation of the progesterone receptor signaling pathway: association with Hepatitis E-related poor pregnancy outcome

Background &amp; Aims: Hepatitis E virus (HEV) infection is associated with high maternal and fetal mortalities. A prospective study was undertaken to evaluate the role of viral and host factors in HEV related pregnancy outcomes. Methods: The study included HEV infected pregnancy cases; acute viral hepatitis (AVH), n = 100 and fulminant hepatic failure (FHF), n = 43, and healthy pregnancy cases, n = 50. HEV genotypes and viremia were studied by nucleotide sequencing and real time PCR, respectively. Progesterone receptor (PR) gene mutations (PROGINS) were studied by PCR, PR expression at the mRNA and protein levels in the placenta were studied by semi-quantitative RT-PCR and immunohistochemistry, respectively. Progesterone induced blocking factor (PIBF) expression was studied by RT-PCR in blood. Serum interleukin-10 (IL-10) and interleukin-12 (IL-12) levels were assayed by ELISA. Results: HEV viral load was significantly higher in FHF than AVH (p &#60;0.001) and in cases with fetal mortality in AVH (p = 0.001) and FHF (p = 0.018). PROGINS were predominant in FHF compared to AVH (p = 0.26) and showed reduced mRNA and protein expression. The risk of fetal mortality in AVH was two times higher (OR, 2.190; CI, 0.303-15.85) and maternal and fetal mortalities in FHF were 4-fold (OR, 4.0; CI, 0.363-44.113) increased in PROGINS carriers. PR and PIBF expression was lower in AVH and even lower in FHF compared to healthy controls. The higher IL-12/IL-10 ratio observed in FHF compared to other groups correlated with fetal mortality in AVH and FHF (p &#60; 0.001). Conclusions: In conclusion, reduced expression of PR and PIBF, a higher IL-12/IL-10 ratio, and a high viral load results in poor pregnancy outcome in Hepatitis E