Comparative histological analysis of cerebellum of representative species of elasmobranchii

SUMMARY: In elasmobranch fishes, variations in gross structural organization of cerebellum has been extensively explored. The basic histological features of cerebellum although conserved in the group but the comparative account on subtle cellular variations is largely underestimated. The present study aims to explore the histological and cellular variations in different layers of cerebellar cortex of the representative elasmobranchs' species belonging to different habitat. Our findings showed that the histological architecture of cerebellar granular layer between the examined species varies noticeably. By and large increase cellular density were observed in all the layers of cerebellum in the representative species of shark compared to ray. The findings were then compared and discussed with reference to their habitat and behavior.

En los peces elasmobranquios, las variaciones en la organización estructural general del cerebelo se han explorado ampliamente. Las características histológicas básicas del cerebelo, aunque se conservan en el grupo, pero la descripción comparativa de las variaciones celulares sutiles es limitada. El presente estudio tiene como objetivo explorar las variaciones histológicas y celulares en diferentes capas de la corteza cerebelosa de las especies representativas de elasmobranquios pertenecientes a diferentes hábitats. Nuestros hallazgos mostraron que la arquitectura histológica de la capa granular del cerebelo entre las especies examinadas varía notablemente. Se observó un gran aumento de la densidad celular en todas las capas del cerebelo en las especies representativas de tiburón en comparación con la raya. Luego, los hallazgos se compararon y discutieron con referencia a su hábitat y comportamiento.

Lip and oral cavity cancers (C00-C06) from a mega city of Pakistan: Ten-year data from the Dow Cancer Registry.

OBJECTIVE: The objective of this report is to provide an overview of lip and oral cavity cancer in Karachi, the largest city in Pakistan. METHODS: This study was undertaken at the Dow Cancer Registry. During 2010-2019, all patients who were residents of Karachi who had been diagnosed with lip and oral cavity cancer were registered and recruited for this study. The data were entered in SPSS and MS Excel sheets to investigate frequencies, age-standardized-rates (ASR) and other clinicopathological parameters. The data from our study were compared with the ASR of lip and oral cavity cancer from selected Asian countries. RESULTS: During the defined period, 22,858 cancer cases were registered. Of these, 4,400 (19.2%, ASR 28.0) were lip and oral cavity cancer (the most common type found in males and the second most common type in females), of which 2,986 (67.8%) were found in males, while 1,414 (32.1%) were reported in females. Squamous cell carcinoma was the most common type (97.7%). Most tumours were moderately differentiated (67.7%) followed by poorly (16.6%), and well differentiated (15.7%). CONCLUSION: We report an alarmingly high ASR of lip and oral cavity cancer in Karachi as compared to Pakistan as a whole and other Asian countries.

HBV S antigen evolution in the backdrop of HDV infection affects epitope processing and presentation.

INTRODUCTION: HBV can evolve under selection pressure exerted by drugs and/or host immunity, resulting in accumulation of escape mutations that can affect the drug or the immune activity. Hepatitis delta virus (HDV) coinfection is also known to exert selection pressure on HBV, which leads to selective amplification of certain mutations, especially in genes that are required for HDV pathogenesis, such as HBsAg. However, little is known about the function of these mutations on HBV or HDV life cycle. The purpose of this study is to determine mutations selectively amplified in the backdrop of HDV, and how these mutations affect processing of CD4- and CD8-T cell epitopes. METHODS: HBsAg was successfully amplified from 49/50 HBV mono- and 36/50 coinfected samples. The sequences were used to identify mutations specific to each study group, followed by an in silico analysis to determine the effect of these mutations on (1) proteasomal degradation, (2) MHC-I and MHC-II biding, and (3) processing of T-cell epitopes. RESULTS: HBV-HDV coinfected sequences exhibited certain unique mutations in HBsAg genes. Some of these mutations affected the generation of proteasomal sites, binding of HBsAg epitopes to MHC-I and -II ligands, and subsequent generation of T- cell epitopes. CONCLUSION: These observations suggest that HBV selectively amplifies certain mutations in the backdrop of HDV coinfection. Selective amplification of these mutations at certain strategic locations might not only enable HBV to counteract the inhibitory effects of HDV on HBV replication but also facilitate its survival by escaping the immune response.

Common Cancers in Karachi, Pakistan: 2010-2019 Cancer Data from the Dow Cancer Registry.

OBJECTIVES: To present 2010-2019 cancer data from the Dow Cancer Registry representing all districts of Karachi (~17.4 million). METHODS: The study was conducted at the Dow University of Health Sciences. After ethical approval, the Dow Cancer Registry was established at the largest government-run diagnostic and reference center of Karachi (Dow Labs). All cancers registered during 2010-2019 were analyzed. Patients >18years of age were labeled as adults while those with ages ≤18years were classified as children/young adults. RESULTS: During 2010-2019, a total of 22,858 cancers were registered. Of these, 9,112(39.9%) cancers were diagnosed in males while 13,746(60.1%) in females. Incidence rates for all cancers (all ages) were 108/1,00,000 for males and 188.6/1,00,000 for females. In adult males, cancer of lip and oral cavity was the most frequently diagnosed cancer (33.6%), followed by non-melanoma -skin-cancer (NMSC) (7.2%), oesophagus (6.8%), colorectum (6.7%) and stomach (4.9%). In adult females, breast cancer was the most frequently recorded malignancy (53.2%), followed by cancers of lip and oral cavity (10.4%), oesophagus (5.3%), colorectum (3.3%) and NMSC (3%). In children, most common malignancy was that of brain and nervous system (15.3%), followed by Hodgkin's lymphoma (14.2%), colorectum (8.1%), endocrine-&-related organs (8%) and non-Hodgkin's lymphoma (7.8%). CONCLUSION: Cancers of lip and oral cavity and breast cancer were the most common malignancies in males and females respectively. In paediatric group, cancers of brain and nervous system were most common. Alarmingly, Karachi males have highest ASR of cancers of lip and oral cavity compared to any other city of Pakistan.

Pan-Cancer Multiomics Analysis of TC2N Gene Suggests its Important Role(s) in Tumourigenesis of Many Cancers.

BACKGROUND: Role of TC2N in carcinogenesis has been largely unfathomed until recently when it was identified as a novel oncogene in lung cancer. Subsequently, a tumour suppressor role of TC2N was reported in breast cancer. It is therefore highly relevant to investigate TC2N molecular partners/mechanisms on a larger scale including a wider range of tumour types. METHODS: We investigated TC2N mRNA expression, its promoter methylation levels, effects of TC2N transcription on overall patient survival, somatic mutations in TC2N gene and correlation between TC2N mRNA expression and other cancer genes in pan-cancer by using data available from the Cancer Genome Atlas (TCGA) and the Genotype Tissue Expression (GTEx) databases. RESULTS: TC2N mRNA expression was differentially regulated in 9/33 TCGA tumour types. Of these 9 tumours, 5 tumour types (cholangiocarcinoma, ovarian-serous-cystadenocarcinoma, rectal-adenocarcinoma, stomach-adenocarcinoma and thymoma) had significantly higher TC2N mRNA expression while 4 (pheochromocytoma-and-paraganglioma, skin-cutaneous-melanoma, thyroid-carcinoma and uterine-carcinosarcoma) had significantly lower TC2N mRNA expression compared to matched and normal controls. TC2N promoter was hypermethylated in most cancers while hypomethylated in head-and-neck-squamous-cell-carcinoma and kidney-renal-clear-cell carcinoma. TC2N transcription was positively correlated with transcription of several other cancer genes including genes from Myc, cell-cycle, Nrf2, Wnt, PI3K, Hippo, Notch, TGFß and RAS/RTK pathways. Poor prognosis was associated with higher TC2N mRNA levels in pancreatic-adenocarcinoma and brain-lower-grade-glioma and lower TC2N mRNA levels in kidney-renal-clear-cell-carcinoma, mesothelioma, sarcoma and skin-cutaneous melanoma. Functional protein partners of TC2N were identified as STX2, SMEK1, SMEK2, STXBP5, SCARA5, MMRN1, CATSPER2, CATSPERB, CLEC4M and STAB2. Many of these proteins are key players in carcinogenesis of various cancers. Highest pathogenic somatic mutation rates in TC2N were found in skin-cutaneous-melanoma, uterine-corpus-endometrial-carcinoma, colon-endocervical-adenocarcinoma, bladder-urothelial-carcinoma and breast-invasive-carcinoma. CONCLUSION: Our findings unravel several un-explored avenues related to the role of TC2N in tumourigenesis of several cancers, suggesting TC2N as an important player and a potential candidate for tumour-therapy.

Non-albicans Candida species: Emergence of neglected pathogens among population of Karachi.

Candida albicans was considered as the principal cause of opportunistic candidiasis but nowadays, neglected non-albicans Candida (NAC) species are evolving as more virulent and drug resistant strains. This research was intended to assess pervasiveness of candidiasis mainly caused by NAC species in Karachi city. A total of 562 clinical isolates of Candida spp. collected during the period of one year were identified by microscopic as well as morphological (germ tube formation, characteristics on CHROM agar and Corn meal agar) and Biochemical (sugar assimilation and fermentation) characteristics. Doubtful species were further identified by using Remel RapIDTM yeast plus kit. The results were statistically analyzed by SPSS 16.0 version software. Isolated strains of candida revealed slight predominance of C. albicans (54.5%) over non- albicans Candida species (45.5%). Among NAC species, C. tropicalis and C. glabrata were isolated as the predominant species. These clinical species were procured mainly from urine samples of females (73.7%) of age group 20-30 years. No significant correlations exist between Candida species and their months of isolation as well as their isolation from different districts of Karachi. Emergence of NAC species may predict an upcoming threat in health care facilities and hence, require prompt management and accurate identification to suggest empirical antifungal therapy.

Bergenin loaded gum xanthan stabilized silver nanoparticles suppress synovial inflammation through modulation of the immune response and oxidative stress in adjuvant induced arthritic rats.

Bergenin (BG) is a naturally occurring C-glycoside with demonstrated anti-arthritic potential. Its therapeutic efficacy is compromised due to its lower absorption and instability at neutral-basic pH. The present study reports fabrication of gum xanthan (GX) stabilized silver nanoparticles (AgNPs) with BG for anti-arthritic activity in a CFA-induced arthritis model targeting ROS, cytokines and TLR expression. NPs were characterized through UV-vis, zetasizer, FT-IR and AFM. Oral administration of BG loaded NPs (1 mg kg-1) exhibited potent anti-arthritic activity with a minimal arthritic score, mild to moderate paw tissue swelling, reduced degenerative changes along with mild articular changes and less influx of inflammatory cells in macroscopic X-ray and histological examination. Administration of BG and its NPs suppressed the levels of reactive oxygen species (ROS) significantly as compared to the arthritic control group. Moreover, increased production of O2˙- in human neutrophils, stimulated by opsonized zymosan (OZ) and phorbol-12-myristate-13-acetate (PMA) was also suppressed. BG and its loaded NPs were revealed to antagonize the oxidative stress via interference with the NADPH oxidase metabolic pathway. Their anti-oxidant activity was further assessed by their inhibitory effect against TLR (TRL-2 & -4) and cytokine (IL-1ß, IL-6 and TNF-α) production. The current investigation validates GX stabilized AgNPs as stable and promising multi-targeted therapeutic nano-cargo for BG delivery with efficient treatment of RA.

Estimation of uncertainty measurement - A prerequisite of ISO1589 accreditation for clinical laboratories.

OBJECTIVE: To estimate relative expanded uncertainty measurement of routine clinical chemistry analytes for international organisation for standardisation 15189 accreditation. METHODS: This cross-sectional study was conducted at Dow International Medical College, Karachi, from September 2013 to May 2014. During the process of international organisation for standardisation 15189 accreditation, measurement uncertainty was estimated for 13 clinical chemistry analytes using top-down approach. Relative combined uncertainty of each analyte was calculated by combining uncertainties of imprecision, bias and calibrators. Results of estimated imprecision, bias and expanded uncertainties were observed for allowable imprecision, bias and total analytical error for the respective analyte.. RESULTS: Uncertainties of imprecision were found within acceptable limits for all analytes except total protein (2.4% vs. 1.3%). Uncertainties of bias of all analytes were found within allowable limits. Relative expended uncertainties of all analytes were found acceptable except total protein (4.7%vs 3.63%). CONCLUSIONS: The approach used to estimate the measurement uncertainty may be found simple and feasible by clinical laboratories interested in getting the relevant accreditation.

Comparison Of Ziehl-Neelsen Based Light Microscopy With Led Fluorescent Microscopy For Tuberculosis Diagnosis: An Insight From A Limited Resource-High Burden setting.

BACKGROUND: Microscopy is the most widely used tool for Tuberculosis screening. Conventionally, Ziehl-Neelsen (ZN) staining has been the widely used for staining Acid-Fast Bacilli (AFB) but with the advent of Fluorescent staining, Auramine O stain is now being adapted as the preferred method for setups with high workload as it has the advantage of being less laborious, since bacteria fluoresce in front of a dark background and are easier to count. This study was performed to compare the efficiency of the two methods in a high-burden, limited resource setting to see the magnitude of diagnostic accuracy between ZN and Fluorescent Microscopy, using culture as the standard.. METHODS: Altogether 987 culturally confirmed cases were considered from the period 36 months during January 2011 to December 2013 and data were compiled from the records maintained at the Provincial Tuberculosis Reference Laboratory at Ojha Institute of Chest Diseases, Dow University of Health Sciences, Karachi. The results from 523 cases examined using ZN and 464 cases using Fluorescent staining method were compared for diagnostic accuracy on the basis of Mycobacterial culture results. Smears are prepared from the clinical samples obtained from presumptive tuberculosis patients. RESULTS: The results of ZN method showed 94.23% [95% CI 91.32-96.39%] sensitivity and 84.91% [95% CI 78.38-90.08%] specificity. While FM showed a sensitivity of 97.15% [95% CI 94.82-98.63%] and specificity of 83.19% [95% CI 74.99-89.56%].. CONCLUSIONS: The results showed that Fluorescent microscopy was slightly more sensitive than ZN light Microscopy, while specificity of both the methods were comparable.

False negativity in AFB Smear microscopy: An insight into the caveats of the most widely used screening tool for tuberculosis.

OBJECTIVE: To study the ratio of false negativity in sputum samples in diagnostic smears received for acid fast bacilli smear microscopy. METHODS: The retrospective cross-sectional study was conducted at the Ojha Institute of Chest Diseases, Karachi, and comprised specimens for microscopy and culture from presumptive tuberculosis patients for 38 months starting from November 2010.All laboratory investigations had been done as per the National Tuberculosis Control Programme guidelines. Statistical analyses were performed on MedCalc and Social statistics calculators, and Open Epi software. RESULTS: TOf the 2,158 specimens, 1,316(60.98%) were of men and 842(39.02%) of women (male-to-female ratio: 1.56:1). Besides, 843(39.06%) were smear-negative, of which 99(11.74%) were false negative. Of the 1,275(96.88%) men whose age was reported, 808(63.37%) were aged 19-45 years, whereas out of the 792(94.06) women whose age was known, 517(65.28%) were in that age group. Microscopic outcomes were significantly related to gender (p<0.001). CONCLUSIONS: Smear microscopy cannot be solely relied upon for diagnosis and its results must be correlated with additional clinical information and other diagnostics due to considerable amount of false negativity, especially in female population.

Cancer patterns in Karachi (all districts), Pakistan: First results (2010-2015) from a Pathology based cancer registry of the largest government-run diagnostic and reference center of Karachi.

National level population-based cancer data have never been published from Pakistan in seven decades since independence (1947). Therefore, generation of high-quality regional data becomes highly relevant. Cancer data for the period of 2010-2015 representing the population from all districts of Karachi (14.6 million) are presented herein. After institutional approval (Ref no. IRB-459/DUHS/-14), a Pathology based cancer registry was established at the largest government-run diagnostic and reference center of Karachi. During 2010-2015, a total of 13,508 cancers (including 686 non-melanoma-skin-cancers (NMSC)) were diagnosed. Of these, 5665 (41.9%) were in males while 7843 (58.1%) were in females. Incidence rates for all cancers (excluding NMSC) were 66.7 per 100,000 (crude) and 105.1 per 100,000 (ASR) for males and 112.0 per 100,000 (crude) and 175.8 per 100,000 (ASR) for females. In males, cancer of lip and oral cavity was the most frequently diagnosed cancer (30.8%, ASR 33.1), followed by NMSC (7.7%, ASR 9.5) and colorectum (7%, ASR 7.3). In females, breast cancer was the most frequently recorded malignancy (49.5%, ASR 87.9), followed by lip and oral cavity (11.2%, ASR 22.0) and oesophagus (5.6%, ASR 10.7). We report that Karachi has the highest incidence of cancers of breast, lip and oral cavity, oesophagus and larynx in females and cancer of lip and oral cavity and larynx (2nd only to Turkey) in males compared to any of the Asian populations. Notably, incidence of tobacco associated cancers is very high in Karachi, demanding urgent attention by relevant authorities to address the un-controlled and drastically high consumption of various forms of tobacco in the city.

DIFFERENCES IN CULTURAL YIELD OF MYCOBACTERIUM TUBERCULOSIS ON MEDIA PREPARED USING COMMERCIAL AND HOUSEHOLD EGGS.

BACKGROUND: Mycobacterial culture is considered as the gold standard for TB diagnosis. It is performed on egg-based media using commercially available eggs to grow Mycobacteria from clinical samples. These eggs are known to contain high concentration of antibiotics, including fluoroquinolones, given to chicken to prevent early mortality. This study was performed to compare Mycobacterial growth on media prepared from commercial and antibiotic free household eggs. METHODS: Sputum samples from negative (No bacilli in 100 oil immersion field), scanty (1-9 AFB in 100 fields), 1+ (10-99 bacilli per field), 2+ (1-10 bacilli per field) and 3+ (>10 bacilli per field) were inoculated dually on Ogawa medium prepared from commercial and household eggs. Tubes were inspected every fourth day for the appearance of colonies till 60 days. Data tabulations and statistical analysis (F test for variation and unpaired Student's t test) were performed on Microsoft Excel. RESULTS: One microscopically negative sample showed growth on media prepared from household eggs, while all were negative on that prepared from commercial eggs. There were significant differences in time to culture positivity for samples graded 1+ (p = 0.02), 2+ (p = 0.002) and 3+ (p = 0.0003). CONCLUSION: Commercial eggs containing antibiotics can be a source of false negativity in cultures especially in microscopically negative samples. This can be of special concern in HIV patients who have high smear negativity. It is therefore important to either develop provision of antibiotic free eggs for media preparation or to develop and validate other laboratory investigations for smear negative TB patients.

In vitro evaluation of antimicrobial effect of miswak against common oral pathogens.

BACKGROUND AND OBJECTIVE: Miswak is a natural tooth cleaning tool which is being used in many parts of the world since ancient times. It is known to be useful in prevention of dental caries. But still it is not used as frequently as other oral hygiene tools. This research was designed to scientifically establish antimicrobial effect of miswak in vitro against common oral pathogens. METHODS: This was a cross-sectional study involving 100 health care workers. This research was carried out in Microbiology section of Dow Diagnostic Research & Reference Laboratory. A questionnaire was designed to test oral hygiene habits of study subjects. Oral swabs were taken and microorganisms were identified by standard bacteriological methods. Test material included four different types of miswaks i.e. (1) root of the peelu (Salvadora persica) tree (in packing) (2) root of the peelu tree (without packing) (3) stem of the peelu tree & (4) stem of the neem (Azadirechta indica) tree. These miswaks were tested against three different types of microorganisms isolated from oral swabs: Staphylococcus aureus, Streptococcus mutans & Candida albicans by agar diffusion method. Inhibition zone was measured after 24 hrs of incubation at 37(o)C. RESULTS: Among the miswaks used, root of the peelu tree in both packing and without packing exhibited strong antimicrobial effect against all three tested microorganisms. However miswak taken from the stem of the peelu and neem tree did not show any antimicrobial activity against all three types of the tested microorganisms. CONCLUSION: Miswak taken from the root of the peelu tree exhibited antimicrobial activity against all the common oral pathogens and could be a good oral hygiene tool in combating dental caries.

Prevalence and clinical profile of insulin resistance in young women of poly cystic ovary syndrome: A study from Pakistan.

OBJECTIVE: The aim was to estimate the prevalence of Insulin Resistance (IR) in Poly Cystic Ovary Syndrome (PCOS) and analyze its clinical parameters. METHODOLOGY: This observational study was conducted at Dow University Hospital during June 2011 till May 2012. Patients of PCOS were selected, an anthropometric measurement, examination and fasting blood test for sugar (FBS) and insulin was performed. Data was collected on pre designed questionnaire, was analyzed by SPSS version 16. RESULTS: Forty-six cases of PCOS were included in the study. Prevalence of IR was 34.78%. Mean age of patients was 23.72 ± 4.37 years. Waist Hip Ratio (WHR) was raised in 42 (91.30%), acanthosis was found in 26(56.50%), impaired FBS was seen in 9 (19.6%) and raised fasting insulin in 16 (34.8%) patients. There was significant association between acanthosis and WHR (0.044) and between acanthosis and FBS (0.008). Correlation studies between parameters showed a significant correlation between Waist & Hips (0.93), similarly Waist & WHR showed positive correlation (0.59), at p< 0.01. Significant positive correlation was also found between waist and FBS (0.32) and FBS & WHR (0.378). CONCLUSION: Acanthosis nigrican, raised WHR and FBS are significant parameters for insulin resistance in cases of Poly Cystic Ovary Syndrome (PCOS).