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Freshwater fish stocks are being exposed to increasing threats as a result of fisheries and aquaculture practices. Integrating genetic knowledge into fisheries and aquaculture management is becoming increasingly important in order to ensure the sustainability of species. So, I used SSR markers to evaluate the pattern of genetic variability in Silver Carp populations (175 samples) from five different sites of River Chenab, Pakistan. DNA was isolated and processed for analysis. There were no scoring errors related to large allele, no stuttering bands, and no null allele. The mean values of number of alleles, allelic richness, effective number of alleles, observed (Ho) and expected (He) heterozygosites, 1-Ho/He, inbreeding coefficient, pairwise population differentiation, and the gene flow provided data indicating loss of genetic diversity of silver carp in River Chenab (Pakistan). Reasons are overhunting, pollution, inbreeding, and poor control measures.

Diclofenac sodium (DCF) is a non-steroidal anti-inflammatory drug mainly used as an analgesic, arthritic and anti-rheumatic. This study deals with the degradation of diclofenac by photo catalytic-based advanced oxidation processes. The solar reactor system is comprised of quartz and borosilicate tubes for the absorption of the solar rays and transmission to the pollutant sample. Artificial UV lamp and solar rays have been applied to activate the ZnO catalyst, thereby generating highly oxidizing species. These species initiate the degradation process of the drug, which results in intermediates that finally dissociate into carbon dioxide and water. The degradation rate has been analyzed by composition analysis using high performance liquid chromatography. TOC and COD tests have also been conducted for degraded samples. ZnO catalyst loading was tested from 0.1 gm/L to 1 gm/L and the degradation rate showed a rising trend up to 0.250 gm/L, but further increase in loading resulted in a drop in degradation. Similarly, degradation is higher in acidic condition as compared to neutral or basic pH condition. The results showed a higher degradation rate for UV lamp irradiation as compared to the solar system. Moreover, TOC and COD reduction results were also found to be higher for UV lamp photo catalysis.

Objective: Cardiovascular diseases are 1.7 times more prevalent in patients with diabetes mellitus. The aim behind this study was to examine the correlation of glycosylated hemoglobin and complexity of coronary artery disease among middle aged population with diabetes mellitus Patients and Methods: This was a clinical prospective hospital-based comparative study carried out in the department of the cardiology department of Isra University Hospital Hyderabad for 10 months. 153 participants with diabetes mellitus having age ≥45 years both male and female and undergoing their first coronary angiography were included. Data were entered and analyzed by using Statistical Package for the Social Sciences version 21.0. Results: A total of 153 patients were finally analysed. Patients were divided into two groups; group I (HbA1c 6.5%, impaired). The mean age was slightly higher in group II 54.60 as compared to group I 53.65 years, respectively. The Syntax score was correlated with HbA1c levels in patients older than 45 years of age (r = 0.001; p >0.05). Moreover, the higher HbA1c levels were observed in every three categories of Syntax score but they were insignificantly associated with each other in patients with diabetes mellitus and having age more than 45 years. Conclusion: The HbA1c is insignificantly correlated with the complexity of coronary artery disease in diabetic patients having age more than 45 years. While only two factors, hypertension and increased triglycerides are significantly different among the HbA1c groups.

Oral squamous cell carcinoma (OSCC) is the most prevalent subtype of head and neck cancers that has a disproportionately high incidence and mortality in developing countries in South-Central Asia, including Pakistan. The standard treatment for OSCC generally consists of surgery combined with radiotherapy or chemotherapy; however, the overall survival rates have not improved, and only ~50% of patients survive the disease. Therefore, identifying and validating new drug targets can provide new therapeutic options for improved treatment and better prognosis for OSCC patients. The present study aimed to develop an organoid model from patient-isolated primary tumour cells and determine their drug sensitivity profile coupled with mutational analysis. New combinations of targeted drugs with synergistic activity in oral cancer cells were identified through high-throughput screening followed by validation of their activity in oral cancer cells in 2D monolayer and 3D organoid cultures. Results indicate a synergistic activity of proteasome (PR-171) and survivin (YM155) inhibitors in all tested patient-derived tumour cells and four oral cancer cell lines. Moreover, chitosan-based nanocapsules harboring these drugs showed therapeutic efficacy comparable to free drug combinations, indicating a potential for efficient and targeted delivery. Next-generation sequencing data identified missense mutations in eleven cancer-related genes that encode BRAF, EGFR, KRAS, NRAS, HRAS, MEK1, PIK3CA, PTEN, NOTCH1, TP53 and TERT. The ongoing research aims to correlate the drug sensitivity data with the mutational profile of patient-derived tumour cells to find novel targets. In conclusion, we have developed 3D organoids from patient-derived primary tumours and utilized them to validate the synergistic combination between screen-identified proteasome and survivin inhibitors. Our preclinical study provides a rationale for further evaluation of the novel drug combination in vivo and in clinical settings. Citation Format: Muhammad Furqan, Iffat Aleem, Muhammad Tahseen, Raza Hussain, Asif Loya, Muhammad Tariq Mahmood, Philippe Gautier, Kevin Myant, Saira Saleem, Amir Faisal. Proteasome and survivin inhibitors synergize to inhibit the growth of oral cancer organoids. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4940.

Background Forkhead box P3 (FOXP3) is a specific marker for immunosuppressive regulatory T (T-reg) cells. T-regs and an immunosuppressive enzyme, indoleamine 2,3-dioxygenase (IDO), are associated with advanced disease in cancer. Aim To evaluate the co-expression of FOXP3 and IDO in triple negative breast cancer (TNBC) with respect to hormone-positive breast cancer patients from Pakistan. Methods Immunohistochemistry was performed to analyze the expression of FOXP3, IDO, estrogen receptor, progesterone receptor, and human epidermal growth factor receptor on tissues of breast cancer patients (n = 100): Hormone-positive breast cancer (n = 51) and TNBC (n = 49). A total of 100 patients were characterized as FOXP3 negative vs positive and further categorized based on low, medium, and high IDO expression score. Univariate and multivariate logistic regression models were used. Results Out of 100 breast tumors, 25% expressed FOXP3 positive T-regs. A significant co-expression of FOXP3 and IDO was observed among patients with TNBC (P = 0.01) compared to those with hormone-positive breast cancer. Two variables were identified as significant independent risk factors for FOXP3 positive: IDO expression high (adjusted odds ratio (AOR) 5.90; 95% confidence interval (CI): 1.22-28.64; P = 0.03) and TNBC (AOR 2.80; 95%CI: 0.96-7.95; P = 0.05). Conclusion Our data showed that FOXP3 positive cells might be associated with high expression of IDO in TNBC patients. FOXP3 and IDO co-expression may also suggest its involvement in disease, and evaluation of FOXP3 and IDO expression in TNBC patients may offer a new therapeutic option.

Background: Tumors use several immunosuppressive mechanisms to evade immune destruction. Cyclooxygenase-2 (COX-2) expression may be a driver of immunosuppression in breast cancer, but the mechanisms involved remain elusive. COX-2 expression induces the expression of indoleamine 2,3 dioxygenase (IDO) in tumor cells. IDO is an immunosuppressive enzyme which is involved in tumor immune escape mechanisms in breast cancer. Our aim was to evaluate the association between COX-2 and IDO expression to find evidence of immunosuppression in Pakistani breast cancer patients. Methods: Immunohistochemical analysis was performed to evaluate the expression of COX-2, IDO, estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2) on formalin-fixed paraffin-embedded breast cancer tissues of 100 patients. Univariable and multivariable logistic regression model was used to identify the independent risk factors of COX-2. Results: A total of 100 patients were included with a mean age and standard deviation of 48.28 ± 11.83. A significant association was observed among COX-2, IDO, ER, PR and tumor grade. In multivariable analysis, three variables were identified as significant independent risk factors for high COX-2: IDO expression high; [adjusted odds ratio (AOR) 6.51; 95% confidence interval (CI) (2.00-21.20), p=0.001], ER; [AOR 5.62; 95% CI (1.80-17.84), p=0.002] and age [AOR 1.04; 95% CI (1.00-1.10), p=0.05] respectively. Conclusion: Our data showed that high IDO expression is associated with high COX-2 expression in Pakistani breast cancer patients. The co-expression of both enzymes may suggest their role in disease pathogenesis. Hence the concurrent targeting of COX-2 and IDO may be a promising therapy for breast cancer.

Kashif Asghar,1 Asif Loya,2 Iftikhar Ali Rana,2 Muhammad Tahseen,2 Muhammad Ishaq,2 Asim Farooq,3 Muhammad Abu Bakar,4 Iqra Masood5 1Department of Basic Sciences, Shaukat Khanum Memorial Cancer Hospital and Research Centre (SKMCH & RC), Lahore, Pakistan; 2Department of Pathology, Shaukat Khanum Memorial Cancer Hospital and Research Centre (SKMCH & RC), Lahore, Pakistan; 3Department of Biomedical Engineering & Sciences, School of Mechanical & Manufacturing Engineering, National University of Sciences & Technology (NUST), Islamabad, Pakistan; 4Department of Cancer Registry and Clinical Data Management, Shaukat Khanum Memorial Cancer Hospital and Research Centre (SKMCH & RC), Lahore, Pakistan; 5Department of Clinical Research, Shaukat Khanum Memorial Cancer Hospital and Research Centre (SKMCH & RC), Lahore, Pakistan Background: Immune dysfunction in breast cancer patients is well established. Indoleamine 2,3-dioxygenase (IDO) is an immunosuppressive enzyme that is linked with progression of cancer. IDO is overexpressed in triple-negative breast cancer (TNBC) cases. Materials and methods: We conducted the first study to analyze IDO expression and overall survival in breast cancer cases in Pakistan. Expression of IDO, estrogen receptor (ER), progesterone receptor (PR), and human EGF receptor 2 (HER2) was evaluated by immunohistochemistry. Formalin-fixed paraffin-embedded breast cancer tissues of 100 (TNBC, n=49 and non-TNBC, n=51) patientswere obtained from Shaukat Khanum Memorial Cancer Hospital and Research Centre. IDO expression was analyzed in association with clinicopathological features and overall survival. A total of 100 patients were classified based on the ordinal IDO score variables as low, medium, and high. In addition, overall mean age and SD of patients was 48.28±11.82. Results: Immunohistochemical analysis showed that high IDO was observed in the TNBC patients (65.3%) compared to that in the non-TNBC patients (33.3%). Multivariable analyses showed that TNBC was an independent risk factor for high IDO expression. Overall survival was also significantly associated with IDO score. Conclusion: Our study showed that IDO protein expression is higher in TNBC patients (P

The critical state of circulatory collapse and hypoperfusion that results in end-organ damage has been called shock. Carcinoid crisis is a rare cause of shock, which is difficult to identify in the presence of infection. It occurs due to the release of vasoactive amines into the systemic circulation following an inciting event. In the presence of a neuroendocrine tumor, carcinoid crisis should be suspected in case of resistant shock. Herein, we report a rare case of carcinoid crisis, in which a quick response to the somatostatin analog therapy, octreotide, was helpful to confirm the diagnosis.

Carbonate apatite/hydroxyapatite (CO3Ap/HAP) additive was obtained by calcination of wasted chicken bones at 900°C. Intermolecular attraction exists between CO3Ap/HAP additive and blended polysulfone (PSF) polymer. Electron dispersive X-ray (EDX) and FTIR analysis were carried out to check the elemental composition and bonding chemistry of prepared additive. The instantaneous demixing process generated consistent finger-like networks in CO3Ap/HAP/PSF-based composite membranes while sponge-like structure was shown by PSF as revealed by SEM images. The increase in weight % of additive loading is also confirmed by EDX analysis. Furthermore, the interaction mechanism of CO3Ap/HAP additive with polysulfone medium was analyzed by FTIR exploration. The water absorption experiment defined a 93% expansion in hydrophilic performance. Change in porosity occurs with additive loading and pure water permeation flux improved up to 11 times. Approximately, antifouling results revealed that 87% of water flux was recovered after treating with a protein solution, whereas a 30% improvement in antifouling capability in case of bovine serum albumin solution occurred. In vitro cytotoxicity, and clotting times study was carried out to evaluate virulent behavior and anticoagulation activity of formulated membranes.

The aim of the study is to identify cornulin (CRNN) protein expression associated with advancement of tongue squamous cell carcinoma (TSCC). A comparison of addictive (containing potential carcinogens) versus non-addiction causative agents was expected to allow detection of differences in CRNN expression associated with TSCC. Bespoke tissue microarrays (TMAs) were prepared and immunohistochemistry (IHC) performed to determine the changes in CRNN expression in epithelial cells of node-negative (pN-), node-positive (pN+) TSCC and non-cancer patients’ oral tissues. TMAs were validated by performing IHC on whole diagnostic tissues. Chi-square test or Fisher’s-exact tests were used to establish significant expression differences. Analogous analyses were performed for biomarkers previously associated with TSCC, namely collagen I alpha 2 (COL1A2) and decorin (DCN) to compare the significance of CRNN. Keratinisation and its level (low, extensive) were studied in relation to CRNN so that the extent of squamous differentiation could better be assessed. IHC immunoreactive score (IRS) clustered the patients based on weak/moderate (Low (IRS ≤ +3)) or strong (High (IRS ≥ +4)) expression groups. A low expression was observed in a larger number of patients in control proteins COL1A2 (77.3%), DCN (87.5%) and target protein CRNN (52.3%), respectively. Low CRNN expression was observed in TSCC where nodes were involved (pN+: mean 1.4 ± 2.1) (p = 0.248). Keratinisation (%) was low (0% ≤ 50%) in 42.2% and extensive (1% ≥ 50.0%) in 57.8% patients. In conclusion, our study suggested that Low CRNN expression was associated with grade and lymph node metastasis in TSCC. CRNN expression is independent of addiction, however potentially carcinogenic addictive substances might be aiding in the disease progression.

Background The aim of this pilot study was to identify proteins associated with advancement of colon cancer (CC). Methods A quantitative proteomics approach was used to determine the global changes in the proteome of primary colon cancer from patients with non-cancer normal colon (NC), non-adenomatous colon polyp (NAP), non-metastatic tumor (CC NM) and metastatic tumor (CC M) tissues, to identify up- and down-regulated proteins. Total protein was extracted from each biopsy, trypsin-digested, iTRAQ-labeled and the resulting peptides separated using strong cation exchange (SCX) and reverse-phase (RP) chromatography on-line to electrospray ionization mass spectrometry (ESI-MS). Results Database searching of the MS/MS data resulted in the identification of 2777 proteins which were clustered into groups associated with disease progression. Proteins which were changed in all disease stages including benign, and hence indicative of the earliest molecular perturbations, were strongly associated with spliceosomal activity, cell cycle division, and stromal and cytoskeleton disruption reflecting increased proliferation and expansion into the surrounding healthy tissue. Those proteins changed in cancer stages but not in benign, were linked to inflammation/immune response, loss of cell adhesion, mitochondrial function and autophagy, demonstrating early evidence of cells within the nutrient-poor solid mass either undergoing cell death or adjusting for survival. Caveolin-1, which decreased and Matrix metalloproteinase-9, which increased through the three disease stages compared to normal tissue, was selected to validate the proteomics results, but significant patient-to-patient variation obfuscated interpretation so corroborated the contradictory observations made by others. Conclusion Nevertheless, the study has provided significant insights into CC stage progression for further investigation.

A rapid response system (RRS) is a program designed to respond in a timely, organized, and comprehensive manner to a patient’s urgent unmet medical need within a healthcare facility. The goals of the rapid response team (RRT) are to restore homeostasis, prevent further physiologic deterioration, and establish an optimal environment of care. RRTs are now in widespread use in the US because of the Joint Commission’s national patient safety goals, which required that healthcare organizations improve recognition and response to changes in a patient’s condition. Recent meta-analyses have now concluded that RRT is effective in reducing the incidence of cardiac arrests within hospitals. There is still controversy, however, on the impact of RRT on ultimate clinical outcomes, including mortality. In this chapter, we review the history and evolution of RRTs, rationale for its existence, its impact on patient outcomes, and current controversies.

Aims: A sub-population of cells named cancer stem cells (CSCs) that initiate and promote tumour growth have been demonstrated to exist in several malignancies including colon carcinoma. The objective of our pilot study was to isolate CD133+CD26+CD44+ CSCs from patient colon tumours, culture spheres or organoids and observe their proliferation in primary cultures. Parallel cultures of non-cancer controls from colon normal lining and nonadenomatous polyps were set up. Methods: Magnetic activated cell sorting was used to isolate CD133+CD26+CD44+ cell populations followed by primary cell culturing under stem cell culture conditions. Number, cells/organoid and daughter generations of organoids were calculated using phase contrast microscope. Trypan blue exclusion method was used to test the viability of the cells. Results: Both colon tumour and colon non-adenomatous polyp formed floating organoids in suspension; however non-adenomatous polyp cultures did not show self-renewal properties for more than 1 passage. Normal colon singlecell suspension did not create organoids. Metastatic colon tumours rapidly produce cancer cell organoids in less than 24 hours in larger numbers compared to non-metastatic colon tumours (1-3 weeks). Metastatic colon tumour organoids have the ability for proliferation for upto five daughter generations in primary culture compared to three generations for those grown from non-metastatic tumours. Conclusions: This in vitro CSC organoid model will help study colon cancer biology, in particular providing a valuable source of primary cell-derived tissue for studying personalized molecular profiling using ‘omics strategies to direct therapeutic intervention.

Background Women harboring BRCA1/2 germline mutations have high lifetime risk of developing breast/ovarian cancer. The recommendation to pursue BRCA1/2 testing is based on patient’s family history of breast/ovarian cancer, age of disease-onset and/or pathologic parameters of breast tumors. Here, we investigated if diagnosis of triple-negative breast cancer (TNBC) independently increases risk of carrying a BRCA1/2 mutation in Pakistan. Methods Five hundred and twenty-three breast cancer patients including 237 diagnosed ≤ 30 years of age and 286 with a family history of breast/ovarian cancer were screened for BRCA1/2 small-range mutations and large genomic rearrangements. Immunohistochemical analyses were performed at one center. Univariate and multiple logistic regression models were used to investigate possible differences in prevalence of BRCA1/2 mutations according to patient and tumor characteristics. Results Thirty-seven percent of patients presented with TNBC. The prevalence of BRCA1 mutations was higher in patients with TNBC than non-TNBC (37 % vs. 10 %, P

Background: Epidermal squamous cell carcinoma (SCC) is a common malignancy in Pakistan. We hypothesize that it is characterized by higher frequency of p53 genetic mutations and increased AgNOR values compared with squamous cell papilloma (SCP) and basal cell carcinoma (BCC). Experimental design: To test our hypothesis, 140 skin biopsies (including 20 normal skin, 20 SCP, 20 BCC and 80 SCC samples of various grades) were examined for p53 mutations using immunohistochemistry (IHC) and polymerase chain reaction (PCR). AgNOR staining was used for histological determination of AgNOR index. Results: Both markers were undetectable in normal skin and were low in SCP. They were upregulated in BCC and SCC. PCR experiments revealed p53 mutations in 70% and 96.25% of BCC and SCC, respectively. Higher AgNOR values were seen in SCC than in BCC (mean AgNOR count = 5.81 ± 31 and 8.36 ± 19; percentage of AgNOR was 43.5% and 53% in BCC and SCC, respectively). Finally, p53 IHC score was found to be related to the AgNOR index in the histological grading of BCC and SCC (r = +0.983, p

Nosocomial infections caused by Acinetobacter species (Spp.) is an emerging threat in health care setups especially intensive care units (ICU). The objective of this observational study was to determine the pattern of Acinetobacter infections and its association with length 6f stay in patients admitted to our medical ICU from January to August 2011.All patients above 16 years of age with stay of more than 48 hours were checked for any development of new infections not present or. incubating at the time of admission. Nosocomial infections were documented in the light of clinical findings and lab results. Data was analysed using statistical software SPSS 15.0.A total of 146 patients had a stay of at least 48 hours; frequency of nosocomial infection was 30.8% out of which 57.8% were Acinetobacter infections. Respiratory system was most commonly involved. Acinetobacter Spp showed high resistance (96.2%) to penicillins, cephalosporins and even extended spectrum antibiotics including carbepenems, quinolones and piperacillin plus tazobactam. Extended drug resistance was seen in 92.3% isolates; while we found high susceptibility to tigecycline (88.5%) and polymyxins (100%). Acinetobacter Spp. infected patients had mean length of stay (LOS) of 12.92 days when compared to patients with other hosocomial infections and no infection with mean LOS of 7.05 days (p=0.05) and 4.86 days (p=0.00) respectively.Acinetobacter Spp infections increase with longer duration of stay in ICU. Emergence of multi-drug and extended-drug resistant Acinetobacter Spp is alarming and overwhelming at this rate for already stretched out health system with its economic and health implications.

Recently a sub-population of cells with stem cell characteristics, reported to be associated with initiation, growth, spread and recurrence, has been identified in several solid tumors including oral tongue squamous cell carcinoma (OTSCC). The aim of our pilot study was to isolate CD44+ cancer stem cells from primary cultures of OTSCC and neck node Level I (node-I) biopsies, grow cell spheres and observe their characteristics in primary cultures. Parallel cultures of hyperplastic lesions of tongue (non-cancer) were set up as a control. Immunohistochemistry was used to detect CD44/CD24 expression and magnetic activated cell sorting to isolate CD44+ cell populations followed by primary cell culturing. Both OTSCC and node-I biopsies produced floating spheres in suspension, however those grown in hyperplastic and node-I primary cultures did not exhibit self-renewal properties. Lymph node metastatic OTSCC, express higher CD44/CD24 levels, produce cancer cell spheres in larger number and rapidly (24 hours) compared to node negative OTSCC (1 week) and non-cancer specimens (3 weeks). In addition, metastatic OTSCC have the capacity for proliferation for up to three generations in primary culture. This in vitro system will be used to study cancer stem cell behavior, therapeutic drug screening and optimization of radiation dose for elimination of resistant cancer cells. Electronic supplementary material The online version of this article (doi:10.1186/s12935-014-0143-3) contains supplementary material, which is available to authorized users.