Ductal variant of prostate adenocarcinoma harbor Xenotropic murine leukemia virus related virus (XMRV) infection: a novel finding in subtype of prostate cancer.

OBJECTIVE: Xenotropic murine leukemia virus related virus (XMRV), is the first gammaretrovirus identified a decade ago, in human tissue bearing adenocarcinoma of prostate, followed by several researches documenting little or no prevalence of XMRV in prostate cancer samples. However, the status of XMRV within subtype of prostate adenocarcinoma has not been investigated yet. In this study, we investigated the relationship between XMRV and broad spectrum morphological entities of prostate adenocarcinoma, including acinar, ductal and other rare subtypes. MATERIAL AND METHODS: The prevalence of XMRV DNA in different histological subtypes of prostate adenocarcinoma was examined after characterizing the tumors into groups, using formalin-fixed, paraffin-embedded tissue samples from newly diagnosed prostate adenocarcinomas and archival prostate cancer tissue from our XMRV case control analysis. Broad-spectrum XMRV DNA amplification was performed by end-point polymerase chain reaction, using commercially available primer set. RESULTS: The study included 100 patients with prostate cancer. XMRV DNA was detected in 4 of 8 (50%) ductal adenocarcinomas, exhibiting papillary and cribriform histological features. XMRV DNA was not detected in any other variant of adenocarcinoma including acinar (0/91) and mucinous carcinomas (0/1). Majority of XMRV positive cases were biologically aggressive and present cancer at an early age upon diagnosis. CONCLUSION: Ductal adenocarcinomas demonstrate a significant association of XMRV DNA while other histological variants of prostate adenocarcinoma seem unrelated to XMRV infection.

Differential Expression of Cytokeratin 13 in Non-Neoplastic, Dysplastic and Neoplastic Oral Mucosa in a High Risk Pakistani Population.

BACKGROUND: Gradual loss of cytokeratin 13 (CK13) may be linked with the severity of dysplastic changes and transformation to malignancy. In this study we assessed the differential expression of CK13 in normal, hyperplastic, dysplastic and cancerous oral mucosa. MATERIALS AND METHODS: A total of 93 oral biopsies were collected during the 2011-2014 period. The biopsies were characterized as normal (19), hyperplastic (21), severely dysplastic/carcinoma in situ (16) and invasive oral squamous cell carcinoma (OSCC) (37) after morphological assessment. Formalin fixed paraffin embedded sections were stained with a monoclonal antibody against CK13 using the Envision technique. Immunohistochemically stained slides were then analyzed for CK13 expression. RESULTS: CK13 was consistently and diffusely expressed in all normal and hyperplastic tissue biopsies from oral mucosa. Severely dysplastic/carcinoma in situ biopsies showed complete loss in 50% of cases, while in the remaining 50% expression was very focal and weak. OSCC cases showed complete or near complete loss of CK13 in all cases. Few cases showed weak expression in keratin pearls only. CONCLUSIONS: This study validates the utility of CK13 IHC as a useful immunohistochemical marker in routine diagnostic practice to make distinction between non-neoplastic from dysplastic and neoplastic (malignant) oral lesions.

Ductal and Acinar Adenocarcinoma of Prostate: Morphological and Immunohistochemical Characterization.

OBJECTIVES: We sought to characterize the ductal and acinar subtype of prostate adenocarcinoma using hematoxylin and eosin (H&E) staining and an immunohistochemical antibody cocktail. We also investigated the clinical features, prostate-specific antigen (PSA) levels, and biological aggressiveness of these tumors. METHODS: We utilized tumor bearing prostate biopsies, obtained between 2010 and 2014 from Dow Diagnostic Research and Reference Laboratory, to identify cases of prostatic ductal and acinar adenocarcinoma using routine H&E and immunohistochemical staining. The immunohistochemical antibody cocktail 34ßE12/p63/AMACR was used for staining. The association of clinicopathological variables including patient's age at diagnosis, Gleason score, and PSA levels before surgery was retrospectively analyzed. RESULTS: A total of 10 ductal and 140 non-ductal cases were identified. Ductal cases were predominantly high grade with advanced histopathological features (90%; p=0.030). Marked elevation in PSA level was also reported in most cases. No other significant statistical difference was observed. CONCLUSIONS: Pathological and immunohistochemical examination could be used to characterize ductal and acinar adenocarcinoma of the prostate. Ductal adenocarcinoma of the prostate is a rare subtype of prostate carcinoma and is be more likely to present with advanced grade cancer suggesting that timely detection of the disease is vital.

Review of diagnostic techniques of hepatic fibrosis.

With increasing innovations aimed at the treatment of chronic liver disease (CLD), the precise staging of liver fibrosis is important to help establish efficacious management of disease activity in each patient. The development of biomarkers capable of non-invasive staging of fibrosis in the liver is challenging as fibrogenesis is a part of the normal wound healing response. There is an array of non-invasive methods, including serum biomarker assays, and imaging techniques such as transient elastography. The degree of implementation of non-invasive diagnostic tests for liver fibrosis differs all over the world, and still remains limited. Liver biopsy so far is the mainstay of diagnosing hepatic fibrosis. Precise staging of liver fibrosis is essential in management of patients. This review provides a systematic overview of various techniques, as well as both approaches based on direct and indirect biomarkers to stage fibrosis, and covers recent studies related to hepatic fibrosis.

Glial Fibrillary Acidic Protein (GFAP) as a Mesenchymal marker of Early Hepatic Stellate Cells Activation in Liver Fibrosis in Chronic Hepatitis C Infection.

OBJECTIVE: This study aims to determine expression of Glial Fibrillary Acidic Protein and of Alpha Smooth Muscle Actin (α-SMA) in hepatic stellate cells of CHC cases and their association with stage of fibrosis. METHODS: The study was conducted at Ziauddin University, Clifton Campus during the year 2010-2012. Sixty Chronic Hepatitis C cases were immmunostained using anti α-SMA antibody and anti-GFAP antibody. Semi quantitative scoring in pericentral, periportal and perisinusoidal area of each case was done to assess immunoexpression of each marker. Results : Immunoexpression of GFAP showed significant association with α-SMA. GFAP expression was inversely correlated with progression of fibrosis. Conclusion : GFAP could represent a useful marker for early hepatic stellate cells activation. Follow up biopsies showing decline in GFAP levels may help identify the target group requiring aggressive therapy.

Successful intravaginal culture of human embryos for the first time in Pakistan--an experience at the Sindh Institute of Reproductive Medicine, Karachi.

Approximately 10-15% of the married couples remain childless. Conventional in vitro fertilisation (IVF) has been the treatment option for most of these cases. However, it is expensive and only available to a small fraction of the infertile population. The Sindh Institute of Reproductive Medicine (SIRM), Karachi, Pakistan, has introduced a new method of IVF for the first time in Pakistan. Intravaginal culture (IVC) is a simple, reliable and cost-effective alternative to IVF. The procedure can be performed in an office set-up with minor capital equipment. Here, we report a case of a subfertile woman treated by IVC, which resulted in successful fertilisation of the eggs (fertilisation rate 71.42%) and embryo development, with an intrauterine pregnancy. Since this is the first successful case of IVC in Pakistan, comparative success rate and take-home baby rate have yet to be established.

Cryopreservation of embryos by vitrification at a private sector reproductive medicine facility in Karachi.

OBJECTIVE: To assess the survival of freezing cleaved human embryos through vitrification. METHODS: The prospective study was conducted at the Karachi-based Sindh Institute of Reproductive Medicine between June 2008 and June 2009. The cryopreservation of embryos being a new technology in Pakistan, only 19 couples, picked through convenience sampling, comprised the study population. The couples were treated for infertility by in virto fertilisation (IVF) or intracytoplasmic sperm injection (ICSI); 125 surplus embryos were vitrified. Subsequently, 15 embryos were thawed, and transferred in a controlled cycle. SPSS version 11 was used for statistical analysis. RESULT: After the surplus embryos were vitrified and subsequently thawed and transferred, the survival of the embryos was assessed by the number of blastomeres that were intact. The overall embryo survival rate was (14/15) 93.33%. CONCLUSION: Vitrification is a simple procedure that requires less time and is likely to become safer and more cost-effective with time. Survival rate after thawing and preserving is high, but comparative success rates in terms of pregnancy and taking-home-baby rates are yet to be established in Pakistan.

Clinical results of the intra-cytoplasmic sperm injection with surgically retrieved sperms in azoospermic men at Sindh Institute of Reproductive Medicine, Karachi.

OBJECTIVE: To analyse the result of intra-cytoplasmic sperm injection (ICSI) by using surgically retrieved sperms in the azoospermic male. METHODS: Eight (9 cycles) azoospermic men were given the intra-cytoplasmic sperum injection after sperms were collected through surgical retrieval either by percutaneous epididymal sperm aspiration (PESA) or testicular sperm extraction (TESE) at the Sindh Institute of Reproductive Medicine (SIRM), Karachi, which is an assisted reproductive technology centre. Fertilisation, cleavage and pregnancy rates were calculated in their spouses with surgical sperm collection (SSC) and intra-cytoplasmic sperm injection. The study related to the period between September 2007 and February 2009. Statistical analysis were done using SPSS version 11.0. RESULT: After the intra-cytoplasmic sperm injection procedure, fertilisation rate of 72.72% and cleavage rate of 78.12% were achieved in the eight patients. A total of 3 (33.33%) clinical pregnancies were obtained through the transfer of embryo from surgical sperm collection in the azoospermic men. CONCLUSION: Intra-cytoplasmic sperm injection with surgical sperm collection proved to be the only successful treatment for men with azoospermia. It gave 33.33% chances of fatherhood in men who were previously labelled infertile. Results were found to be promising and are expected to improve with time and experience.

Frequency of high risk human papillomavirus types in squamous cell carcinoma of cervix among women.

OBJECTIVE: To determine the frequency of high-risk subtypes of Human papillomavirus (HPV) 16, 18, 31 and 45 in squamous cell carcinoma of cervix (SCC) among women. METHODS: A descriptive study was conducted in the department of Molecular Genetics Laboratory, Ziauddin University Hospital, North Nazimabad, Karachi, Pakistan. A total of fifty formalin-fixed paraffin embedded samples (blocks) of SCC of cervix were collected from two Pathology laboratories through convenience sampling. These blocks were analyzed for presence of HPV and its subtype. DNA was extracted by QIAamp DNA Kit and amplification was done by Polymerase chain reaction (PCR). General primers were used for HPV DNA and HPV genotyping for 16, 18, 31 and 45 was done by using a standard kit. RESULTS: HPV was detected in nine (18%) out of fifty paraffin embedded tissues of squamous cell carcinoma of cervix. Out of the 9 cases that were positive, five (55.6%) were infected with HPV16, while in the rest of the positive samples, the genotype could not be identified in four (44.4%). HPV was not determined in majority (82%) of the samples. CONCLUSION: This study concluded that HPV might not be the major cause of SCC of cervix. There could be other causes among women leading to prevalentHPV types that the study did not look into due to limitations.

Frequency of hypothyroidism in patients of beta-thalassaemia.

OBJECTIVE: To determine the frequency of hypothyroidism in patients suffering from homozygous beta-thalassaemia. METHODS: This descriptive study included 70 diagnosed thalassaemia major patients aged 5-14 years. Demographic data as well as history of blood transfusion and chelation therapy was collected. Random blood samples were drawn and thyroid profile (serum thyroxine [T4], triiodothyronine [T3] and thyroid stimulating hormone concentrations [TSH]) was done by enzyme-linked immunosorbent assay (ELISA). Primary hypothyroidism was defined by a TSH level > 4 microIU/ml. Results were analysed by descriptive statistical methods. RESULTS: Primary hypothyroidism was seen in 18 (25.7%) patients. Of these, 17 had normal T4 levels with elevated TSH levels consistent with a diagnosis of compensated primary hypothyroidism whereas only one patient showed a decreased T4 level with elevated TSH (uncompensated primary hypothyroidism). Mean age of hypothyroid patients was 9.2 +/- 2.6 years. Frequency of hypothyroidism was associated with increased serum ferritin levels. CONCLUSION: Primary hypothyroidism occurs in a significant proportion of thalassaemia major patients in the absence of obvious clinical signs of hypothyroidism. Regular follow-up for early detection and timely treatment of such complications could improve the quality of life of these patients.

Prophylactic and therapeutic human papillomavirus vaccine: a breakthrough for women health.

Human Papilloma Virus (HPV) associated cervical cancer is amongst the ten most common cancers in women in the developing world. New vaccines markedly reduce the economic burden of diseases on a country's health system. Two types of HPV vaccines are available; prophylactic and therapeutic. Prophylactic vaccines target (Late protein) L1 and to a lesser extent L2, which stimulate neutralizing antibody response and are recommended to individuals prior to infection. Therapeutic vaccines are recommended to individuals who are already exposed to HPV infection and these induce a specific T-cell mediated immune response targeted to (Early protein) E6 and E7 oncogenes for the eradication of existing lesion. Several studies have been done which show that these vaccines are highly effective. This review highlights the pathogenesis of HPV infection, importance of HPV vaccines, recent advances and few clinical trials on prophylactic and therapeutic HPV vaccine, their economic impact and the need to implement the vaccination programmes in the developing world.

Association of Helicobacter pylori with carcinoma of stomach.

OBJECTIVE: To note the association of Helicobacter pylori in patients having carcinoma of stomach. METHODS: A descriptive study was carried out at the Department of Histopathology, Ziauddin Medical University, Karachi from April 1992 to May 1998. Histological evaluation of 50 cases of carcinoma of stomach was compared with 50 cases each of chronic gastritis and histological normal gastric mucosa. Only those cases of carcinoma of stomach were included that contained sufficient non-neoplastic mucosa in addition to tumour tissue. Three glass slides with serial sections of each case of carcinoma of stomach, chronic gastritis and normal gastric mucosa were freshly cut and stained with H&E, PAS and Giemsa stains. All slides were examined by light microscopy. RESULTS: Helicobacter pylori were identified in 35 cases (70%) of carcinoma of stomach, in 42 cases (84%) of chronic gastritis, and in 12 cases (24%) of normal gastric mucosa. The presence of H. pylori in cases of carcinoma of stomach and chronic gastritis was highly significant (P < 0.001) as compared to normal gastric mucosa. Chronic gastritis was observed in the non-neoplastic mucosa in 48 cases (96%) with carcinoma of stomach. Of 50 cases with carcinoma of the stomach, intestinal type of carcinoma was found in 30 cases (70%), and diffuse type in 15 cases (30%). No significant difference was noted in the prevalence of H. pylori between intestinal type (69%) and diffuse type (71%) gastric carcinoma. Significant Helicobacter pylori associated chronic gastritis was observed in intestinal type (94%) and diffuse type (100%) of gastric carcinoma. The prevalence of H. pylori was insignificant in the presence or absence of mucosal atrophy and intestinal metaplasia in both types of gastric carcinoma. CONCLUSION: A significant number of H. pylori were found in patients of carcinoma of stomach. Both intestinal and diffuse types of gastric carcinoma showed strong association with H. pylori. Chronic gastritis appears to be the background lesion while atrophy and intestinal metaplasia indicate long term infection.