Unveiling the Antibiotic Susceptibility and Antimicrobial Potential of Bacteria from Human Breast Milk of Pakistani Women: An Exploratory Study.

Background: Human life quality and expectancy have increased dramatically over the past 5 decades because of improvements in nutrition and antibiotic's usage fighting against infectious diseases. Yet, it was soon revealed that the microbes adapted to develop resistance to any of the drugs that were used. Recently, there is great concern that commensal bacteria from food and the gastrointestinal tract of humans and animals could act as a reservoir for antibiotic resistance genes. Methodology. This study was intended for evaluating the phenotypic antibiotic resistance/sensitivity profiles of probiotic bacteria from human breast milk and evaluating the inhibitory effect of the probiotic bacteria against both Gram-negative and Gram-positive bacteria. Results: The results point out that some of the isolated bacteria were resistant to diverse antibiotics including gentamycin, imipenem, trimethoprim sulfamethoxazole, and nalidixic acid. Susceptibility profile to certain antibiotics like vancomycin, tetracycline, ofloxacin, chloramphenicol, streptomycin, rifampicin, and bacitracin was also observed. The antimicrobial qualities of cell-free supernatants of some probiotic bacteria inhibited the growth of indicator bacteria. Also, antimicrobial properties of the probiotic bacteria from the present study attributed to the production of organic acid, bacterial adhesion to hydrocarbons (BATH), salt aggregation, coaggregation with pathogens, and bacteriocin production. Some isolated bacteria from human milk displayed higher hydrophobicity in addition to intrinsic probiotic properties like Gram-positive classification, catalase-negative activity, resistance to gastric juice (pH 2), and bile salt (0.3%) concentration. Conclusion: This study has added to the data of the antibiotic and antimicrobial activity of some probiotic bacteria from some samples of Pakistani women breast milk. Probiotic bacteria are usually considered to decrease gastrointestinal tract diseases by adhering to the gut epithelial and reducing population of pathogens and in the case of Streptococcus lactarius MB622 and Streptococcus salivarius MB620 in terms of hydrophobicity and exclusion of indicator pathogenic strains.

Isolation and Characterization of Lactobacillus crispatus, Lactococcus lactis, and Carnobacterium divergens as Potential Probiotic Bacteria from Fermented Black and Green Olives (Olea europaea): An Exploratory Study.

Background: Table olives are becoming well recognized as a source of probiotic bacteria that might be used to create a health-promoting fermented food product by traditional procedures based on the activities of indigenous microbial consortia present in local environments. Methodology. In the present study, the characterization of probiotic bacteria isolated from mince, chunks, and brine of fermented green and black olives (Olea europaea) was done based on morphological, biochemical, and physiological characteristics. Results: Bacterial isolates demonstrated excellent survival abilities at 25, 37, and 45°C and at a variable range of pH. However, the optimum temperature is 37 and the optimum pH is 7 for all three isolates. An antimicrobial susceptibility pattern was found among these isolates through the disc diffusion method. Most of the isolates were susceptible to streptomycin, imipenem, and chloramphenicol, whereas, amoxicillin showed resistance to these isolates, and variable results were recorded for the rest of the antibiotics tested. The growth of the isolates was optimum with the supplementation of 3% NaCl and 0.3% bile salt. The isolated bacteria were able to ferment skimmed milk into yogurt, hence making it capable of producing organic acid. Conclusion: Isolates of Lactobacillus crispatus MB417, Lactococcus lactis MB418 from black olives, and Carnobacterium divergens MB421 from green olives were characterized as potential candidates for use as starter cultures to induce fermentation of other probiotic food products.

Streptococcus lactarius MB622 and Streptococcus salivarius MB620 isolated from human milk reduce chemokine IL-8 production in response to TNF-α in Caco-2 cell line, an exploratory study.

Streptococci are a predominant genera of the human milk microbiome. Among different lactic acid bacteria (LAB) a few Streptococcal strains are also considered as probiotics. Probiotic bacteria are reported to modulate immunity when consumed in adequate amount and bacterial hydrophobicity can be considered as a preliminary experiment for the adhesive capability of probiotic bacteria to the epithelial cells. The present study aimed to investigate the probiotic, hydrophobic and immune modulation property of Streptococcus lactarius MB622 and Streptococcus salivarius MB620, isolated from human milk. S. lactarius MB622 and S. salivarius MB620 displayed higher hydrophobicity (78 % and 59 % respectively) in addition to intrinsic probiotic properties such as gram positive classification, catalase negative activity, resistance to artificially stimulated gastric juice and gastrointestinal bile salt concentration. In conclusion Streptococcus lactarius MB622 and Streptococcus salivarius MB620 isolated from human milk when administered in sufficient amount and for certain duration could be used to reduce inflammation inside the colon by reducing the production of inflammatory booster (IL-8) in diseased state.

Genetic polymorphism in ERCC5 and breast cancer risk.

ERCC5 plays crucial role in excision repair DNA damage induced by UV in NER pathway. Single neuleotide polymorphism in ERCC5 were responsible for different cancers. Therefore, current study evaluated the relationship between ERCC5 (rs1047768 T>C) polymorphism and the risk of breast cancer in Pakistani population. The rs1047768 polymorphism was screened among 175 females including one hundred breast cancer cases and age matched seventy-five healthy controls. Genotyping was performed with Tetra amplification-refractory mutation system (ARMS) PCR and products were observed through electrophoresis. Multivariate logistic regression was used to calculate odds ratio (OR) and 95% confidence interval (95% CI) investigating relationship between genotypes, clinical parameters and risk of breast cancer. Statistical analysis exhibited significant relationship between the TC genotype (OR=7.2, 95% CI=1.5-34.3) and increased breast cancer risk. Moreover, family history (OR=6.25; 95% CI= 2.61-15.00) and late menopause (OR=2.41; 95% CI=1.20-4.83) were found to be breast cancer associated risk factors. In conclusion, ERCC5 (rs1047768 T>C) polymorphism may contribute towards increased risk of breast cancer in Pakistani population.

Survival analysis of breast cancer patients with different treatments: a multicentric clinicopathological study.

OBJECTIVE: To explore and better understand clinic pathological details of breast cancer patients and analyse their survival rate among different treatment groups. METHODS: The prospective cohort, multi-centric study was conducted from September, 2014, to February, 2018, at five hospitals in Rawalpindi and Islamabad, Pakistan, and comprised histo-pathologically confirmed breast cancer cases. Patient characteristics and medical history were collected using a detailed questionnaire. All the subjects were followed up, and information regarding their current health and treatment status was collected. Data was analysed using SPSS 24. RESULTS: There were 347 subjects with a mean age of 44.3±12.2 years and body mass index of 27.9±4.0 kg/m2. Younger age, increased body mass index, consanguinity and family history were major contributing factors in breast cancer development (p<0.05). Overall, 267(77%) had invasive ductal carcinoma and Grade II tumour 234(67%) was more frequent. A total of 221(64%) cases had positive lymph nodes and 97(28%) had metastasis to different body organs. Overall survival analysis showed statistically significant role (p<0.0001) of all treatment options. CONCLUSIONS: Combination of different treatments can provide more promising health outcomes in breast cancer cases.

Factors influencing delayed presentation of breast cancer at a tertiary care hospital in Pakistan.

BACKGROUND: Breast cancer is the leading cause of death despite the continuous development of newer and more effective modalities of treatment for breast cancer. In Asia, Pakistan has the highest rate of breast cancer. Breast cancer treatment shows better prognosis when it is diagnosed at an early stage, but mortality increases significantly with delayed diagnosis and advanced stage of disease. Delay in diagnosis and nonavailability of treatment are the major factors responsible for advanced stage and low survival. AIMS: The objective of our study was to identify the factors responsible for delayed presentation of patients with breast carcinoma. METHODS AND RESULTS: A cross-sectional study using a questionnaire method was conducted at the Foundation University Medical College from January 2015 to December 2016. A total of 89 patients gave consent and were interviewed using a prestructured questionnaire during the study. Age ranged from 25 to 64 years. Majority of patients were in stage T3N1M0 (31.5%). Second most common stage was T4N0M0 (14.6%). Thirteen patients (12.4%) were in stage T3N0M0, and 10 patients (11.2%) were in T3N2M0. Delay ranged from 3 months to more than 1 year; 43.8% presented with delay of 3 to 6 months. The reasons for delay were lack of knowledge about breast cancer (41%), lack of availability of health care services (32.6%), purdah and religious reasons (6.7%), and fear of being diagnosed with cancer (10.1%). CONCLUSION: The main reasons for delay identified in our study were lack of knowledge and availability of appropriate health care facilities. In order to improve outcome of breast cancer, more focus is needed on spreading awareness and improving health care services in rural areas.

Inter-Rater Reliability Of Objective Structured Long Examination Record.

BACKGROUND: Objective Structured Long Examination Record (OSLER) scale was introduced in 1997 by Gleeson to improve the long case examination. There is no psychometric evidence to support reliability of OSLER. This study was done to analyse inter-rater reliability of OSLER. METHODS: Two groups of examiners assessed 105 students in long case examination of their final professional examination, using OSLER scale. Group 1 was composed of actual examiners while Group 2 was mock examiners. Kappa statistic and intraclass correlation coefficient (ICC) were used on SPSS 23 to calculate reliability. RESULTS: Mean score awarded by actual examiners was 55.36 (SD=11.2) whereas mean score by mock examiners was 57.74 (SD=14.1). Cronbach's alpha was 0.586, Kappa was 0.019 whereas inter-rater reliability on ICC was 0.413. CONCLUSIONS: Although OSLER is a practical modification of long case examination with good validity, the scale needs to be more structured to improve its reliability.

Role Of Tuberculin Test As A Diagnostic Tool For Tuberculosis.

BACKGROUND: Childhood Tuberculosis remains one of the major public health concerns in developing countries like Pakistan and is responsible for high rates of morbidity and mortality in children. Although tuberculin skin test is very commonly used by physicians all over the world, its interpretation always remains difficult and challenging. The objective of this study was to determine the frequency of positive tuberculin skin test in vaccinated and unvaccinated children suffering from tuberculosis. METHODS: This Cross-sectional study was conducted in the department of Paediatric Ayub Teaching Hospital, Abbottabad from 1st February 2015 to 30th April 2016. A total of 150 patients were observed in this study. Children of either gender who were aged 1-15 years admitted in ward with tuberculosis were included in the study by using nonprobability convenient sampling technique. We injected 0.1 ml (10 units) of tuberculin purified protein derivative (PPD) into the anterior surface of the forearm and induration was read at 72 hours after administration. Data was entered and analysed using SPSS version 10. RESULTS: Out of 150 children, 84 (56%) were males and 66 (44%) were females. The mean age was 7.8±3.84 years. Of these 75 (50%) were vaccinated and 75 (50%) were unvaccinated. In vaccinated Group 5.3% children had positive tuberculin skin test while in unvaccinated Group 2.7% children had positive tuberculin skin test and this difference was found statistically insignificant (pvalue= 0.40). Pulmonary TB was the diagnosis in 67 (44.7%), TBM in 65 (43.3%), abdominal TB in 7 (4.7%), disseminated TB in 4 (2.7%) and military TB in 7 (4.7%) patients. CONCLUSIONS: The positivity of tuberculin skin test in vaccinated and unvaccinated children suffering from tuberculosis was found to be insignificant in our study. We conclude that Tuberculin Skin Test should not be used as a sole diagnostic tool for diagnosing the disease in children of our region..

Teenage jejunoileal diverticulosis.

Presentation of jeujunoileal diverticulosis in young age is virtually unknown. It is associated with middle or old age. It is usually asymptomatic but may present with vague abdominal pain and episodic nausea, vomiting or diarrhoea. It can lead to complications like bleeding, perforation and obstruction. We had 3 cases of jejuno-ileal diverticulosis presenting in their teens. They had resection and anastomosis resulting in improved health.