Histopathological Changes in Small Intestine of Rotavirus Infected Poultry

Background: Rotaviruses are important cause of acute gastroenteritis. Group A and D rotavirus are the predominant enteric viruses groups in birds. Outbreaks of rotavirus may lead to significant economic losses in poultry industry. Rotavirus infection alters the function of the small intestinal epithelium, resulting in diarrhea. Methods: Poultry intestinal samples were collected in 10% formalin and duodenum, jejunum and ileum were processed for histopathological examination by H and E staining. Result: Histopathological changes were noticed in all the three parts of the small intestine namely duodenum, jejunum and ileum in poultry intestinal content sample positive for AvRVD in RT-PCR. Duodenum showed necrosis, desquamation and loss of enterocytes from the villi. The jejunum showed severe disruption of villous architecture with vacuolation and separation of mucosal epithelial layer. Ileum showed a complete loss of enterocytes from the villous surface, congestion at the villous tips and infiltration of lymphocytes throughout the mucosa as well as submucosa.

Relationship of Placental TLR-7 Expression with Cord Blood HBV DNA and Placental HBV DNA

Objective: To determine the role of TLR-7 expression on intrauterine vertical transmission in pregnancy through identification of serum hepatitis B markers in both maternal and umbilical cord blood. Method: Analysis of TLR expression was performed on 38 paraffin block samples of placental tissue acquired from mothers with HBV using TLR immunohistochemical staining. Result: 16 of 38 samples were acquired from mothers aged 26-30 years old. Most of the samples were from primiparous mothers (52.6%). This study found no significant association between TLR-7 expression and HBV DNA in the placenta and cord blood (p=1.000). However, we found a significant association between placental TLR-7 expression and maternal HBV DNA (p=0.034). Meanwhile, placental HBeAg and HBV DNA were not associated with placental TLR-7 expression (p=0.082; p = 1.000). Conclusion: There was no significant association between TLR-7 expression and HBV DNA in the placenta and cord blood, but we found a significant association between TLR-7 expression and maternal HBV DNA. Key word: toll-like receptor (TLR) 7, HBV DNA, umbilical cord, placental, Hepatitis B, intrauterine infection

Immune cell and tumor cell-derived CXCL10 is indicative of immunotherapy response in metastatic melanoma

A T cell-inflamed tumor microenvironment is characterized by the accumulation and local activation of CD8+ T cells and Bat3-lineage dendritic cells, which together are associated with clinical response to anti-programmed cell death protein 1 (anti-PD-1)-based immunotherapy. Preclinical models have demonstrated a crucial role for the chemokine CXCL10 in the recruitment of effector CD8+ T cells into the tumor site, and a chemokine gene signature is also seen in T cell-inflamed tumors from patients. However, the cellular source of CXCL10 in human solid tumors is not known. To identify the cellular source of CXCL10 we analyzed 22 pretreatment biopsy samples of melanoma metastases from patients who subsequently underwent checkpoint blockade immunotherapy. We stained for CD45+ and Sox10+ cells with multiparameter immunofluorescence staining, and RNA in situ hybridization technology was used in concert to identify CXCL10 transcripts. The results were correlated with the expression levels of CXCL10 transcripts from bulk RNA sequencing and the best overall response to immune checkpoint inhibition (anti-PD-1 alone or with anti-CTLA-4) in the same patients. We identified CD45+ cells as the major cellular source for CXCL10 in human melanoma metastases, with additional CXCL10 production seen by Sox10+ cells. Up to 90% of CD45+ cells and up to 69% of Sox10+ cells produced CXCL10 transcripts. The CXCL10 staining result was consistent with the level of CXCL10 expression determined by bulk RNA sequencing. The percentages of CD45+ CXCL10+ cells and Sox10+ CXCL10+ cells independently predicted response (p<0.001). The average number of transcripts per cell correlated with the CD45+ cell infiltrate (R=0.37). Immune cells and melanoma cells produce CXCL10 in human melanoma metastases. Intratumoral CXCL10 is a positive prognostic factor for response to immunotherapy, and the RNAscope technique is achievable using paraffin tissue. Strategies that support effector T cell recruitment via induction of CXCL10 should be considered as a mechanism-based intervention to expand immunotherapy efficacy.

Comparative Study of Coronary Artery Proximal to Myocardial Bridge Segment

Introduction: The muscular segments which overlie the epicardial arteries are termed as myocardial bridges and the artery which travels through them are termed as tunnel arteries. These tunnel arteries get compressed during the systolic compression of the heart, thus partially or completely blocking the blood supply to the corresponding areas. Aim & Objectives: To assess the impact of these myocardial bridges on the proximal segment of the myocardial arteries. Methodology: The present study was cadaveric-based cross-sectional study. A total of 22 hearts which showed the presence of myocardial bridges were collected from two sources namely: cadaver dissections, autopsy. The hearts were clean and numbered. This was followed by fixation, dehydration, clearing, embedding, block formation, section cutting and staining. Result: The present study showed that there is a significant thickening in the tunica intima of the proximal to bridge segment of the coronary artery. The present study also noted that there is a marked thinning of the tunica media of the same segment. Conclusion: The present study concludes that there is a marked hyperplasia in the proximal segment of the myocardial bridges under tunica intima.

Life After Death

Objectives To test the performance characteristics of 69 primary immunohistochemistry antibodies after expiration and compare with fresh primary antibodies wherever possible. Methods A total of 69 expired primary antibodies were evaluated for specificity, background staining, and intensity. An optimal staining result corresponded to a semiquantitatively scored 2+ or 3+ intensity, with intact specificity devoid of moderate or strong background staining. Any deviation from a normal staining pattern was also considered to be a suboptimal result. Results Nearly half of the antibodies studied showed an optimally positive staining result after expiration (34/69, 49.2%). Overall, 10.1% (7/69) of antibodies could be compared with fresh primary antibodies of the same clone with equivalent results. Eight of 69 (11.6%) expired antibodies showed splotchy or granular staining. Conclusions Evidence from this study and previous work point to maintained functionality of a fair number of primary immunohistochemical antibodies after expiration. Decisions about the use of such reagents should be guided by a thorough assessment of functionality by the pathologist rather than a manufacturer-specified deadline. Quality maintenance should imply a sensible balance between histopathologic performance and economics.

The Concordance of Three Diagnostic Test for Malassezia folliculitis using Potassium Hydroxide 20% + Blue-Black Parker Ink, May Grunwald Giemsa, and Potassium Hydroxide 10% + Chicago Sky Blue

Background: Malassezia folliculitis is a pilosebaceous follicular infection disease caused by Malassezia species. There are many misdiagnosed Malassezia folliculitis cases, causing the maladministration of therapy. A routine diagnostic test performed for Malassezia folliculitis cases is the identification of fungal elements (spore) with a microscope using potassium hydroxide, but it has several weaknesses. Purpose: To evaluate the suitability of Malassezia folliculitis diagnostic test using Potassium Hydroxide 20% + Blue-Black Parker Ink, May Grunwald Giemsa, and Potassium Hydroxide 10% + Chicago Sky Blue. Methods: Analytic observational study conducted in the Dermatomycology Division of Dermatology and Venereology outpatient clinic, Dr. Soetomo General Hospital Surabaya. The samples were thirty patients with clinical features of Malassezia folliculitis. The research material was obtained from the body as many as three pieces of papulomoluscoid lesion extracted. The material obtained was then divided into three glass objects for Potassium Hydroxide 20% + Blue-Black Parker Ink, May Grunwald Giemsa, and Potassium Hydroxide 10% + Chicago Sky Blue staining. Result: The identification of spores using Potassium Hydroxide 20% + Blue-Black Parker Ink was 90%, May Grunwald Giemsa was 90%, and Potassium Hydroxide 10% + Chicago Sky Blue was 93% with a value of κ=0.348 and p=0.051. The diagnostic values of May Grunwald Giemsa and Potassium Hydroxide 10% + Chicago Sky Blue were 96.6% sensitivity, 33.3% specificity, 92.9% Positive Predictive Value, and 50 % Negative Predictive Value. Conclusions: There was no significant concordance between May Grunwald Giemsa and Potassium Hydroxide 10% + Chicago Sky Blue with Potassium Hydroxide 20% + Blue-Black Parker Ink in establishing the diagnosis of Malassezia folliculitis. Potassium Hydroxide 20% + Blue-Black Parker Ink is still needed as a routine examination in cases with clinical features of Malassezia folliculitis.

The Effectivity of Artemisia Vulgaris Extract on Cyclin-D1 and Ki-67 Decreased as a Supplementation of Adenocarcinoma Mammae Chemotheraphy (Study on C3H Mice Given AC Chemotheraphy Regimen)

Background: The incidence of breast cancer worldwide is still high. Surgery remains the top choice with other modalities of chemotherapy, radiation, and suplementation such as Artemisia vulgaris (AV). Aim: The study was aimed to demonstrate that administration of AV extract decreased the expression of Cyclin-D1 and the expression of Ki-67 in adenocarcinoma mammae. Method: This study used “Posttest only control group design” on 24 females C3H mice that were randomly selected and divided into four groups: group K (control), P1 (chemotherapy), P2 (extract), and P3 (combination). Adenocarcinoma mammae comes from the inoculation of donor mice. Chemotherapy of Adriamycin 60 mg / m2 and Cyclophosphamide 600 mg / m2 were given in 2 cycles. AV 13 mg (0.2 ml) was given once daily orally. Cyclin-D1expression and Ki-67 expression were evaluated by imunohistochemical staining. Result: Mean of Cyclin-D1expression and Ki-67 expression were found in groups K, P1, P2, P3 were 35,30 + 0,72; 20,38 + 0,67; 33,50 + 0,71; 17,36 + 0,66; and 66,44 + 0,65; 35,40 + 0,65; 64,12 + 0,85; 32,32 + 0,61. The statistical analysis showed that there were significant differences in the expression of Cyclin-D1 between groups of K vs P1, P3 (p = 0,001), P1 vs P2 (p = 0,001), P1 vs P3 (p = 0,045), P2 vs P3 (p = 0,001), and in Ki-67 expression between groups of K vs P1, P3 (p = 0,001), P1 vs P2 (p = 0,001), P1 vs P3 (p = 0,041), P2 vs P3 (p = 0,001). Correlation analysis between Cyclin-D1 expression and Ki-67 expression showed significant correlation (p = 0,030 dan r = 0,914). Conclusion: Artemisia vulgaris is potential as supplementation that can improve the effectivity of AdriamycinCyclophosphamide chemotherapy in terms of decreased expression of Cyclin-D1 and expression of Ki-67 adenocarcinoma mammae of C3H mice.

Kepositifan Induksi Sputum NaCl 3% dan Teknik Broncho Alveolar Lavage pada Pneumocystis Pneumonia

Background: Pneumocystis pneumonia (PCP) is a major cause of morbidity and mortality in patients immunocompromised. The incidence of PCP in HIV are 0.3 cases per 100 person years with mortality 63.6%. The diagnosis of PCP experiencing difficulties because of the causative organism can not be cultured. Several attempts were carried out to obtain a representative sample sputum through induced sputum and bronchoalveolar lavage. This study compared the use of induced sputum and bronchoalveolar lavage (BAL) in the diagnosis of PCP. Methods: From September, 2015, to February, 2016, HIV-positive patients 21 to 65 years old were evaluated at UPIPI ward, Soetomo hospital with suspicion of PCP based on clinical and radiological findings. Sputum induction and BAL was done for Giemsa staining. Result: Thirteen subjects with a mean age of 40, with 11 male (84.6%). All subjects with chief complain shortness of breath and common complain cough with hard to expetorate. Most frequent risk factors was freesex. Mean of subjects received treatment cotrimoxazole is 3.5 days. Six subjects have been treated with ART. Mean of LDH serum was 554.62 ± 376.707 U/l. Interstitial infiltrate was the most frequent radiological pattern (76.9%). Most bronchoscopy examinations showed normal results (84.6%). Both Giemsa staining from induced sputum and BAL showed no positive results. Conclusion: All Giemsa staining from both induced sputum and BAL can not be compared due to no positive result.

Verapamil (VER) Enhances the Cytotoxic Effects of Docetaxel and Vinblastine Combined Therapy Against Non-Small Cell Lung Cancer Cell Lines

Lung cancer is one of the foremost tumor-associated cause of death in the world. Most of the patients with NSCLC possesses an advanced disease at diagnosis, and are thus probable subject for systemic therapy. This study aims to evaluate the cytotoxicity of vinblastine and docetaxel combined therapy for the treatment of NSCLC, as well as verapamil (VER) enhancement of the combined therapy. We conducted P-glycoprotein (P-gp) gene expression, protein expression with RT-PCR and western blot respectively, apoptotic response of the combined therapy with VER is also determined using DAPI staining (%). Result of DAPI staining confirmed combination therapy promotes cell apoptosis to greater extent as compared to each drug alone. Real-time RT-PCR analysis revealed that mdr-1 expression level increased 6 fold with docetaxel (40 nM) and 2 fold with vinblastine (30 nM) after 24 h (p<0.001). Consequently, combination therapy reduced drug-induced up-regulation of mdr-1 significantly (p<0.05). VER with the drug combination increased P-gp expression (p<0.05). These data provide evidence showing combined therapy is a better approach to improve the efficacy of chemotherapy and decreasing drug resistance.

SURVEY, ISOLATION, BIOCHEMICAL CHARACTERIZATION, AND IDENTIFICATION OF HELICOBACTER PYLORI FROM GASTRIC PATIENT BIOPSY

Objective: This study was to isolate and identify the Helicobacter pylori from the biopsy samples of the gastric patient.Methods: Gastric biopsies were collected from the antral region of the gastric patient. Out of 96 patients, 59 males and 37 females in the age group between 11 and 70 years old were selected. A serial dilution of the sample was made. The bacterial colonies were examined on the basis of Gram staining, colony morphology, and biochemical reactions such as catalase, urease, oxidase, nitrate reduction, glycine utilization, growth (different media, different temperature, and salt tolerance), and antibiotic sensitivityResult: From the findings, it was found that 75% (65% of male and 35% of female) have H. pylori infection remaining 25% were not infected. The rate of infection was found to be more in age group 55-65 and less in age group below 25. Among 75% of H. pylori infected patients, 72% are affected with ulcer, 19% with gastric cancer, and 8.3% found to be non-gastric inflammated. Gram staining result declared that the isolated bacteria from the biopsy sample observed to be Gram-negative, spiral shaped rod. Biochemical reports produced positive indication to all the tests.Conclusion: Based on the morphological, staining and biochemical test result, it was confirmed that the isolated bacteria was found to be H. pylori.Gastric cancer is the fourth most common cancer and the second leading cause of cancer-related to death worldwide. In 1994, the international agencyfor research on cancer classified H. pylori as a Class I (definite) carcinogen, as H. pylori infection is considered as an important trigger in the processof carcinogenesis of both types of distal gastric cancer.