Role of Nuclear Morphometry in the Cytologic Evaluation of Benign and Malignant Breast Lesions.

Breast cancer is the most common diagnosed cancer in female over the world. It is the most important cause of 'cancer death' among female. For diagnosis of breast masses the Fine needle aspiration cytology (FNAC) is applied as the primary tool. Though it is an easy, rapid and comparatively minimally invasive and inexpensive procedure for quick diagnosis, it is largely a subjective tool. Computer-assisted image morphometry provides a new influential method for high-precision measurement of nuclear features and can help to differentiate between benign and malignant breast aspirates. Therefore, the aim of study was to evaluate computer assisted semi-automatic nuclear morphometry on cytology of breast lesions to differentiate benign and malignant lesions of breast. Total 70 cases were included in this cross sectional study, performed in the department of Pathology, Mymensingh Medical College, Mymensingh, Bangladesh from March 2019 to February 2021. Morphometric analysis was done on images captured from FNAC slides of the selected cases. Image J Morphometric Software was used for image analysis. Statistical analysis was done by using SPSS 26.0 version. On analysis of morphometry, four nuclear size parameters, two nuclear shape parameters and one nuclear chromasia parameter were measured. The cut-off values with sensitivity and specificity between benign and malignant category for mean nuclear area were 61.54µm², 91.0%, 97.0%, for mean maximum feret diameter 10.89µm, 94.0%, 95.0%, for mean minimum feret diameter 7.71µm, 94.0%, 97.0% and for mean nuclear perimeter 33.32µm, 94.0%, 95.0% respectively. So, in this observation, morphometry is adjunctive tools which may overcome the limitations of inter observer agreement, improve diagnostic accuracy and avoid unnecessary repeat Fine Needle Aspiration (FNA) in breast aspirates.

Vascular Endothelial Growth Factor Expression in Colorectal Adenocarcinoma of Resected Samples and its Association with Histological Grade and Pathological Stage.

Colorectal carcinoma (CRC) is one of the major public health problems. Vascular endothelial growth factor (VEGF) is an effective angiogenic factor and plays a pivotal role in the development, progression and metastasis of CRC. It also could further help in selecting patients of high risk of disease progression for adjuvant therapy. Therefore, this study was undertaken to reveal the importance of angiogenic factor (VEGF) as an adjunctive tool with histologic parameters of CRC. Total 45 cases were included in this cross-sectional observational study, performed in the department of Pathology, Mymensingh Medical College, Mymensingh and Immunohistochemistry was done in the department of Pathology, Bangabandhu Sheikh Mujib Medical University, Dhaka, Bangladesh from March 2019 to February 2021. All the cases were evaluated for VEGF immunohistochemical expression. Majority of cases were low grade adenocarcinoma (86.9%) and most common stage was stage III (48.9%). Among 45 cases collectively 13 (4-negative + 9-weak positive) cases were low for VEGF expression and 32 (22-moderate positive + 10-strong positive) cases were high for VEGF expression. Statistically significant association was found with VEGF expression and increasing tumor stage as well as with lymph node metastasis (p<0.05). VEGF had positive significant correlation with stage (r=0.322; p=0.031) of tumor. However, no correlation with VEGF expression and grade (r=0.219; p=0.149) and other clinicopathological parameters of tumor was seen.

Identification of Genes Encoding Aminoglycoside Modifying Enzymes among Clinical Isolates of Proteus species at a Tertiary Care Hospital in Dhaka, Bangladesh.

Proteus is considered as one of the major opportunistic pathogens liable for nosocomial infections and acquired several resistances to a wide range of antimicrobials such as aminoglycosides. The most common mechanism of aminoglycoside resistance is the inactivation of drugs by modifying enzymes. So, this cross-sectional study was undertaken to investigate the occurrence of aminoglycoside resistance and identify aminoglycoside modifying enzyme (AME) genes among clinical isolates of aminoglycoside resistant Proteus spp. A total of 40 Proteusmirabilis and Proteus vulgaris were isolated in the Department of Microbiology of Dhaka Medical College, Dhaka, Bangladesh from July 2018 to June 2019 of 500 wound swab & pus, urine and blood samples. Disk diffusion test was performed by modified Kirby Bauer method. Minimum inhibitory concentration (MIC) of amikacin was determined by agar dilution method. PCR was used to detect aac(3)-Ia, aac(6')-Ib, ant(4')-IIa, ant(2'')-Ia a and aph(3'')-Ib AMEs genes among aminoglycoside resistant Proteus spp. Sequencing of aac(6')-Ib gene was performed to identify aac(6')-Ib-cr variant. Thirty-two (80%) aminoglycoside resistant isolates were detected during disk-diffusion technique. The marked increase in MIC was observed between 256 - ≥2048µg/ml to amikacin. The most prevalent AME-genes were aac(6')-Ib (37.5%), ant(2'')-Iaa (21.86) followed by ant(4')-IIa(12.5%), aph(3'')-Ib (12.5%) andaac(3)-Ia (9.38%). The most frequent combination was aac(6')-Ib + aac(3)-Ia+ant(2'')-Iaa and aac(6')-Ib + ant(4')-IIa + aph(3'')-Ib(2 strains) followed by aac(6')-Ib + aac(3)-Ia(1 strain). Sequencing of aac(6')-Ib gene in this study did not harbor aac(6')-Ib-cr variant gene. The results of this study provide insight into the presence of high AME-genes among Proteus spp. in Bangladesh.

Prevalence of qnr and aac(6')-Ib-cr Genes in Clinical Isolates of Proteus spp. at a Tertiary Care Hospital in Dhaka, Bangladesh.

Plasmid mediated quinolone resistance (PMQR) has been revealed to play not only a significant role in quinolone resistance but also this drug resistance can spread from one bacterium to another. There is limited data regarding the prevalence of PMQR are available from Bangladesh. So, the aim of this study was to detect the prevalence of qnr and aac(6')-Ib-cr genes among clinical isolates of ciprofloxacin resistant Proteus spp. This cross-sectional study was carried out in the Department of Microbiology of Dhaka Medical College, Dhaka, Bangladesh from July 2018 to June 2019. Fourty (40) Proteus spp. was isolated from 300 culture positive samples. Proteus mirabilis and Proteus vulgaris were identified by culture and biochemical test. Antibiotic susceptibility was performed by disc-diffusion technique. Quinolone resistance genes (qnrA, qnrB, qnrC, qnrD, qnrS and aac(6')-1b-cr) among ciprofloxacin resistant Proteus spp. were detected by PCR. Thirty (75%) ciprofloxacin resistant isolates were detected during disk-diffusion technique. Among them, quinolone resistance genes were found positive 11(36.67%) for aac(6')-Ib-cr, 6(20%) for qnrA, 5(16.67%) for qnrD, 4(13.33%) for qnrS and 3(10%) for qnrB genes. Co-existance of qnrA + aac(6')-Ib-cr and qnrD + qnrS were found in 3(10%) wound swab & pus and urine samples respectively followed by qnrA + qnrB in 2(6.67%) wound swab and pus and qnrA+qnrS in 1(3.33%) urine sample. The results of this study showed presence of high (66.67%) percentage of PMQR genes as well as high (30%) rate of co-carriage of the two genes among Proteus spp. isolates. The incidence of PMQR genes was found to be high which could be due to the increased prescription of fluoroquinolones. Thus, there is a need for rational usage of fluoroquinolones.

Socio-demographic and Clinico-epidemiological Study of Scrub Typhus in A Tertiary Care Hospital of Mymensingh, Bangladesh.

Scrub typhus is one of the leading causes of undifferentiated treatable febrile illness in Asia pacific region. It is grossly under diagnosed in many tropical countries of South Asia including Bangladesh, due to wide range of non-specific clinical presentations, low index of suspicion among clinicians, limited awareness and lack of accurate diagnostic facilities. This cross sectional observational study was conducted at department of Microbiology, Mymensingh Medical College, Mymensingh, Bangladesh from March 2019 to February 2020 enrolling 113 diagnosed cases of scrub typhus by Immunochromatographic test (ICT) and / or Nested PCR to characterize the socio-demographic and clinico-epidemiological features of scrub typhus in Mymensingh area. Majority of the scrub typhus cases came from rural areas (63.83%) and there was a slight female predominance (52.21%). The young (32.74%) and the young-adult age group (28.31%) were mostly affected. Most of the scrub typhus cases were housewives (30.98%), followed by farmers (23.89%) and students (21.23%). All the enrolled cases presented with fever. Other findings were myalgia (76.10%), headache (56.63%), cough (30.97%), vomiting (12.38%) and Respiratory distress (9.73%). Typical eschar of scrub typhus was present only in 9(7.96%) cases and 4(3.53%) patients had rashes on their skin. Few cases (3.53%) had jaundice and 15.96% cases were anaemic. Oliguria (7.96%) and neck rigidity (1.76%) were also documented. Most of the Nested PCR positive scrub typhus cases were documented during late rainy season and beginning of winter months. Findings of the study may offer increased awareness about high burden of scrub typhus as well as heightened suspicion among clinicians for early diagnosis, timely treatment and prevention of complications.

Detection of Antimicrobial Resistance Pattern and ESBL Production among Clinical Isolates of Salmonella Species in Mymensingh, Bangladesh.

The occurrence of antimicrobial resistance in Salmonella enterica serovars (both typhoidal and non-typhoidal Salmonellae) is a major public health problem especially in developing countries, which have been associated with treatment failures. Therefore, the study was undertaken to determine the current antimicrobial resistance pattern and extended spectrum ß-lactamase (ESBL) production among clinical isolates of Salmonella spp. during 2019-2020 in Mymensingh, Bangladesh. In this cross sectional study, 36 Salmonella enterica isolates were obtained from blood and stool culture of suspected 200 enteric fever and 100 gastroenteritis patients attending at Mymensingh Medical College Hospital, Mymensingh, Bangladesh. Isolated Salmonella species were identified by biochemical tests and Polymerase Chain Reaction (PCR). Disk diffusion test was performed by modified Kirby Bauer method. Minimum Inhibitory Concentration (MIC) of ceftriaxone was detected by agar dilution method. Double disk synergy test was used as a screening test for ESBL production. PCR was done for detection of blaTEM, blaSHV and blaCTX-MU genes. The isolates showed 25% resistance to Ceftriaxone and 58.3% to Azithromycin. The highest sensitivity rates were 88.9% to Meropenem and 83.3% to Amikacin. Whereas 6(16.7%) isolates were Multi Drug Resistant (MDR). Eight (8) isolates were confirmed as ESBL producer by DDST. The marked increase in MIC was observed between 8->512µg/ml to ceftriaxone. blaTEM, blaSHV and blaCTX-MU genes were detected in 3, 5 and 8 isolates respectively. In conclusion, the current study observed, higher level of resistance to ceftriaxone and azithromycin. At the same times 22.2% isolates showed ESBL production, which is a cause for concern as it may lead to treatment failure. On the other hand the study also showed the re-emergence of chloramphenicol and Sulfamethoxazole-Trimethoprim sensitivity.

Quick Identification and Differentiation of Proteus spp. by PCR and RFLP Along with Their Antibiotic Resistance Pattern.

Different Proteus species are encountered in human infections and may vary with the type of infections they cause. So, the present study was conducted to detect species of Proteus by PCR and RFLP along with their antibiotic resistance pattern. This cross-sectional study was conducted in the Department of Microbiology of Dhaka Medical College, Dhaka, Bangladesh, from July 2018 to June 2019. A total of 500 wound swab and pus, urine and blood samples were tested for bacterial pathogens. Proteus spp. were identified and differentiated by biochemical test, PCR and RFLP. Antibiotic susceptibility was performed by disc-diffusion technique. Fourty Proteus spp. was isolated from 300 culture positive samples, giving 13.33% prevalence of Proteus infections. Proteus mirabilis and Proteus vulgaris were identified by culture, biochemical test, PCR and RFLP. The results were similar by both methods (biochemical tests and PCR). RFLP of 16S rRNA fragments digested with HaeIII revealed that P. mirabilis consisted of two bands at approximately 110 and 190 bp and P. vulgaris consisted of three bands at approximately 100, 180 and 220 bp. The proportion (80%) of P. mirabilis was more than P. vulgaris. Highest proportion (77.5%) of Proteus spp. was isolated from wound swab and pus followed by urine samples. A significant proportion of Proteus spp. was multidrug resistant (90%) and extensively drug resistant (37.5%). Fosfomycin was found the most sensitive drug followed by imipenem. This study provided an insight into antibiotic resistance pattern of Proteus spp. and showed high level resistance towards commonly used antimicrobial agents. PCR and RFLP may be suitable method to identify and differentiate species of Proteus and to treat them accordingly.