Genetic Characterization of Extended-Spectrum Beta-Lactamase (ESBL) and Metallo-Beta-Lactamase (MBL) Producing Klebsiella pneumoniae from Diabetic Foot Ulcer (DFU).

Background: Antibiotic resistance in common pathogenic bacteria is linked with the genetic makeup. The genetic basis of antibiotic resistance may vary in different species or pathophysiological conditions. Objectives: We studied the antibiotic resistance in Klebsiella pneumonia isolates from DFU in the western Indian population. We also studied the presence of ESBL and MBL mechanisms of antibiotic resistance along with the prevalence of the genes involved in ESBL (TEM ESBL , SHV ESBL , and CTX-M ESBL ) and MBL (NDM-1 bla , KPC bla , OXA-48 bla , and VIM bla ) production. Results: A total of 161 K. pneumoniae isolates were analyzed; among which 50.93% were positive for ESBL and 45.96% were positive for MBL production. Most of the isolates were resistant to antibiotics used in the present study and partially resistant to Imipenem and Amikacin. There was no relation between the antibiotic resistance of the isolates and the production of ESBL or MBL mechanism of antibiotic resistance. Further, TEM ESBL was the most prevalent gene in K. pneumoniae isolates followed by CTX-M ESBL , NDM-1 bla , SHV ESBL , and KPC bla . VIM bla was the least prevalent gene found in K. pneumoniae isolates. There was no difference in the prevalence of the genes with respect to the presence or absence of ESBL and MBL mechanism of resistance. Further, there was no relation between the prevalence of the genes and antibiotic resistance in K. pneumoniae isolates. Conclusion: These results along with the literature review suggest that the prevalence of the genes involved in antibiotic resistance mechanisms are widespread in India and their distribution varies in different studies.

CYP2D6, CYP2E1 Gene Polymorphisms and Gastrointestinal Cancer Risk in Rural Maharashtra: A Hospital Based Case-Control Study.

BACKGROUND: Cytochrome P450 (CYP) is a family phase I metabolizing enzymes important in xenobiotics metabolism. Genetic polymorphisms of CYPs have been comprehensively studied for their association with a range of diseases including cancer risk. In this study we assessed single nucleotide polymorphism (SNP) CYP2D6 and CYP2E1 genes and their role in gastrointestinal (GI) cancer susceptibility in the rural population of Maharashtra. METHODS: Genotyping of CYP2D6*4, CYP2E1*5B, CYP2E1*6, CYP2E1*7B genes among 200 GI cancer cases and equal number of controls was studied by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The Odds ratio (OR) with 95% confidence interval and p-value were evaluated to get the level of association of polymorphisms with risk of GI cancer, where p ≤0.005 was considered as statistically significant. RESULTS: After the analysis of CYP2D6 and CYP2E1 gene polymorphisms, we noticed that CYP2D6*4 (rs3892097) with heterozygous genotype (G/C) showed negative association with GI cancer risk (OR=0.43, 95% CI: 0.25-0.74; p=0.002) and CYP2E1*6 (rs6413432) variant genotype showed positive association (OR=2.85, 95% CI: 1.40-5.81; p=0.003) showed positive association with GI cancer risk in studied population. CONCLUSION: The findings obtained from this study concluded that the polymorphic CYP2D6 was negatively associated; however CYP2E1*6 polymorphism was significantly associated with GI cancer risk in studied population.

Single Nucleotide Polymorphisms in APE1, hOGG1, RAD51 Genes and their Association with Radiotherapy Induced Toxicity among Head and Neck Cancer Patients.

BACKGROUND: Radiotherapy (RT) is a crucial treatment for head and neck cancer however, it causes adverse reactions to the normal tissue and organs adjacent to target tumor. The present study was carried out to investigate possible association of single nucleotide polymorphism in DNA repair genes with toxicity effects of radiotherapy on normal tissue. METHODS: Three hundred and fifty head and neck cancer patients receiving radiotherapy treatment were enrolled in this study. The adverse after effects of radiotherapy on the normal tissue in the form of skin reactions were recorded. Single nucleotide polymorphisms of APE1 (rs1130409), hOGG1 (rs1052133) and Rad51 (rs1801320, rs1801321) genes were studied by polymerase chain reaction-Restriction fragment length polymorphism (PCR-RFLP) and direct DNA sequencing methods and their association with development of severe radio-toxicity effects was evaluated logistic regression analysis. RESULTS: The 172G/T polymorphism of Rad51 was 2.85 times higher and significantly associated with skin reactions (OR=2.85, 95% CI: 1.50-5.41; p=0.001) and severe oral mucositis (OR=4.96, 95% CI: 2.40-10.25; p<0.0001). These results suggested that the polymorphic nature of Rad51 is responsible for risk of radiotherapy adverse effects in HNC patients. The variant 326Cys and heterozygous 326Ser/Cys genotype of hOGG1 was significantly associated with high tumor grade (OR=3.16 95% CI: 1.66-5.99; p=0.0004, and OR=3.97 95% CI: 2.15-7.34; p=<0.0001 respectively). The homozygous variant 172TT genotype of Rad51 showed positive association with poor response of both tumor and nodes towards radiotherapy treatment (p=0.007 and p=0.022). CONCLUSIONS: Interpretation of our results revealed significant association of rs1801321 SNP of Rad51 with development of adverse toxicity reactions in normal tissue of head and neck cancer patients treated with radiotherapy.

Expression of caveolin 1 in oral squamous cell carcinoma.

Context: Caveolin-1 is a surface protein that is a major structural component of caveolae, which are vesicles of the plasma membrane integral to a variety of signal transduction molecules and transport functions. Caveolin-1 is a biomarker undergoing research & studies have shown an increased expression of Cav-1 in the stepwise carcinogenesis from the normal oral mucosa, hyperplastic mucosa, dysplastic mucosa, precancerous lesions to Oral Squamous Cell Carcinoma. In the present study Correlation between Caveolin-1 expression and grade of tumor was established statistically. Aims: To study immunohistochemical expression of Caveolin-1 in Oral Squamous Cell Carcinoma. Settings and Design: Cross sectional study carried out in a tertiary care hospital. Materials and Methods: A total of 90 cases of histopathologically diagnosed oral squamous cell carcinoma was evaluated. Grading of the cases into well, moderate and poorly differentiated carcinomas was done as per WHO guidelines . Margin and lymph node status were evaluated. Anti- Caveolin-1 antibody (E249)- Caveolae marker ab32577 was used in the dilution of 1:100. Results were expressed taking reference of the methodology used by Hung et al 2003. Statistical Analysis Used: Statistical Package for the Social Sciences (SPSS 25.0). Results: Correlation of tumor grade and lymph node metastasis was statistically significant p=0.0006. There was a significant statistical correlation between tumor grade and immunohistochemical expression of Caveolin-1, p- value=0.00. Correlation between Lymph node metastasis and Caveolin-1 was statistically significant, p-value=0.008. Conclusions: Caveolin-1 expression correlates with aggressive tumor behavior and poor prognostic outcome.

Phenotypic and Genotypic Characterization of AmpC Beta-Lactamase in Clinical Isolates of Pseudomonas aeruginosa Findings From a Tertiary Care Hospital.

Background and aim Pseudomonas aeruginosa is an opportunistic pathogen responsible for various healthcare-related infections, which are difficult to treat due to intrinsic and acquired resistance. This study aimed to investigate AmpC ß-lactamase production using phenotypic and genotypic methods in Pseudomonas aeruginosa strains isolated from a tertiary care hospital in Karad, Maharashtra, India. Material and methods Over one year, a descriptive cross-sectional study was conducted at the Department of Microbiology, Krishna Institute Medical Sciences, Krishna Vishwa Vidyapeeth, Karad. Phenotypic detection of AmpC beta-lactamase was performed using the Cefoxitin-Cloxacillin Double-Disc Synergy Test method, and genotypic detection was conducted using conventional polymerase chain reaction (PCR) targeting the bla Pseudomonas-derived cephalosporinases (PDC) and bla cephamycinase (CMY) genes. Results Out of 205 clinical isolates of Pseudomonas aeruginosa, 110 (53.66%) showed AmpC production phenotypically, while 86 (41.95%) were positive genotypically. The blaPDC gene was detected in 36.10% of isolates, and the blaCMY gene in 10.73% of isolates. Conclusions The study findings indicate that AmpC-ß-lactamase stands out as the primary resistance mechanism in strains of Pseudomonas aeruginosa isolated from the hospital. PCR study concluded that blaPDC (36.10 %) was the leading gene responsible for AmpC synthesis among study isolates. Early detection of AmpC beta-lactamase production by employing phenotypic and genotypic methods is crucial for detecting antibiotic resistance. This dual approach enables healthcare professionals to decide on the most effective antibiotics and mitigate the development of resistance.

Genetic Polymorphisms in Glutathione S-Transferase (GST) Gene and Their Correlation with Toxicity of Chemotherapy in Breast Cancer Patients.

BACKGROUND: Glutathione S-Transferase (GST) is a family of phase II metabolizing enzymes contribute to detoxification and elimination of variety of endogenous as well as exogenous xenobiotics including chemotherapeutic agents. The comprehensive knowledge on the impact of genetic polymorphisms in GST) enzyme coding gene will help to understand the clinical outcomes in breast cancer patients treated with either Adriamycin or paclitaxel or combination of both. In this study we attempted to assess the genetic polymorphisms in GSTM1, GSTT1, GSTP1 and their association with Adriamycin and Paclitaxel induced toxicity reactions in breast cancer patients. METHODS: Two hundred BC patients receiving Adriamycin and Paclitaxel chemotherapy were enrolled in this study and chemotherapy induced hematological and non-hematological toxicity reactions were noted. The polymorphisms in GSTM1, GSTP1 and GSTT1 gene were studied by PCR and RFLP analysis. RESULTS: After the univariate analysis of the genetic polymorphisms of GSTM1, GSTP1 and GSTT1 showed that GSTT1 null genotype showed significant association with neutropenia (OR=2.84, 95% CI: 1.06-7.56; p=0.036) in breast cancer patients treated with Adriamycin and GSTT1 null genotype in patients with >1 CINV toxicity confirmed significant correlation (OR=3.75, 95% CI: 1.46-9.59; p=0.005). The genetic polymorphisms of GSTP1 (exon 5) A/G heterozygous genotype was significant in grade >1 toxicity reactions of mucositis (OR=3.22, 95% CI: 1.06-9.71; p=0.037) in breast cancer patients administered with Paclitaxel chemotherapy. CONCLUSION: The findings obtained from this study proposed significant involvement of GSTT1-null genotype in hematological neutropenia toxicity in response to Adriamycin and GSTM1-null genotype showed negative association with non-hematological toxicity (bodyache) in response to Paclitaxel.

Interactive Association of XRCC1, XRCC2, XRCC3, and TP53 Gene Polymorphisms With Gastrointestinal Cancer Risk: Insights From a Hospital-Based Case-Control Study.

BACKGROUND AND AIM: Gastrointestinal (GI) cancer presents a significant worldwide health burden, influenced by a combination of genetic and environmental factors. This study endeavors to explore the combined effects of the XRCC1, XRCC2, XRCC3, and TP53 genes that contribute to the heightened risk of GI cancer, shedding light on their combined influence on cancer susceptibility. MATERIALS AND METHODS: A total of 200 histologically confirmed cases of GI cancer and an equal number of controls were selected to examine genetic polymorphisms within the XRCC1, XRCC2, XRCC3, and TP53 genes using the polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP). Odds ratios (OR) with 95% confidence intervals (CI) were calculated to assess the association of these polymorphisms with GI cancer susceptibility, with statistical significance (p ≤ 0.05). RESULTS: Logistic regression analysis confirmed strong evidence of synergistic interactions among specific variant genotypes. Notably, combinations such as heterozygous Arg/Ser+Ser/Ser genotype of TP53 Arg249Ser polymorphism with Arg/Trp+Trp/Trp genotype of XRCC1 Arg194Trp polymorphism (OR=2.64; 95% CI: 1.35-5.18; p=0.004), Arg/Gln+Gln/Gln genotype of XRCC1 at codon 399 (OR=5.04; 95% CI: 2.81-9.05; p=0.0001), Arg/His and His/His genotypes of XRCC2 Arg188His (OR=2.16; 95% CI: 1.06-4.39; p<0.032), and Thr/Met+Met/Met genotype of XRCC3 Thr242Met (OR=3.48; 95% CI: 1.79-6.77; p=0.0002) showed significant associations with GI cancer risk in the study population. CONCLUSIONS: The findings indicate a notable association between the combined effect of heterozygous variant genotypes of TP53 and variant genotypes of XRCC1, XRCC2, and XRCC3 on GI cancer risk. However, further research with a larger sample size and broad single nucleotide polymorphism (SNP) spectra is necessary to understand the interaction between genetic variations and environmental factors influencing GI cancer susceptibility.

The Effect of CYP2C19*2 (rs4244285) and CYP17 (rs743572) SNPs on Adriamycin and Paclitaxel based Chemotherapy Outcomes in Breast Cancer Patients.

BACKGROUND: Cytochrome P450 (CYP) are phase I metabolizing enzymes involved in detoxification of chemotherapeutic agents. Among the CYP gene family, including CYP1A1, CYP1B1, CYP2C, CYP2D, CYP2E and CYP17, their significance in cancer susceptibility is well established. However, there remains limited understanding regarding the polymorphisms of CYP2C19*2 and CYP17 and their potential correlation with chemotherapy-induced toxicity reactions in breast cancer (BC) patients. In this study we intended to identify the association of CYP2C19*2 and CYP17 gene polymorphisms on drug response as well as toxicity reactions in BC patients undergoing adriamycin/paclitaxel based chemotherapy within Indian population. METHODS: Two hundred BC patients receiving adriamycin and paclitaxel chemotherapy were enrolled in this study and chemotherapy induced hematological and non-hematological toxicity reactions were noted. The polymorphisms of CYP2C19*2 (681G>A) and CYP17 (34T>C) isoforms of cytochrome p 450 gene was studied by PCR and RFLP analysis. RESULTS: The univariate logistic regression analysis revealed significant associations between CYP2C19*2 (681 G>A) polymorphisms with hematological toxicities i.e., anemia (OR=9.77, 95% CI: 2.84-33.52; p=0.0003), neutropenia (OR=5.72, 95% CI: 1.75-18.68; p=0.003), febrile neutropenia (OR=4.29, 95% CI: 1.32-13.87; p=0.014) and thrombocytopenia (OR=5.86, 95% CI: 1.15-29.72); p=0.032) in BC patients. Additionally BC patients treated with adriamycin exhibited significant association between CYP2C19*2 polymorphism with chemotherapy induced nausea and vomiting (CINV) (OR=99.73, 95% CI: 5.70-174.64); p=0.001), fatigue (OR=83.29, 95% CI: 4.77-145.69); p=0.002), bodyache (OR=4.44, 95% CI: 1.24-15.91); p=0.021) and peripheral neuropathy (OR=12.00, 95% CI: 1.80-79.89); p=0.010. Furthermore, the regression analysis indicated an association between CYP17 with body ache (OR=2.77, 95% CI: 1.21-6.34; p=0.015) and peripheral neuropathy (OR=3.90, 95% CI: 1.59-9.53; p=0.002) in BC patients treated with paclitaxel chemotherapy. CONCLUSION: The findings obtained from this study illustrated significant association of CYP2C9*2 (681G>A) polymorphism with adreamicin based chemotherapy induced toxicities and CYP17 (34T>C) polymorphism with paclitaxel induced bodyache and peripheral neuropathy in BC patients.

Influence of (C1236T and C3435T) Polymorphisms of ABCB1 Gene on Chemotherapy Treatment Outcome and Toxicity in Breast Cancer Patients.

BACKGROUND: ATP Binding Cassette Transporters (ABCB1) gene plays an important role in transport of different metabolites and anticancer drugs across the cell membrane. There is lack of knowledge on ABCB1 gene polymorphism and its correlation with Adriamycin or paclitaxel based chemotherapy induced toxicity in breast cancer patients. Therefore in this study, we explored the correlation of ABCB1 polymorphisms gene on response and toxicity in adriamycin and paclitaxel based chemotherapy in breast cancer patients from Indian population. METHODS: Two hundred BC patients receiving Adriamycin and paclitaxel chemotherapy were enrolled in this study and chemotherapy induced hematological and non-hematological toxicity reactions were noted. The polymorphisms in ABCB1 gene (C1236T, C3435T) were studied by PCR and RFLP analysis. RESULTS: The univariate logistic regression analysis showed statistically significant negative association with protective effects of ABCB1 (C3435T) polymorphism with heterozygous genotype (OR=0.34, 95% CI: 0.13-0.89; p=0.027), homozygous variant genotype (OR=0.31, 95% CI: 0.10-0.99; p=0.049) and combined C/T+T/T genotypes (OR=0.33, 95% CI: 0.13-0.79; p=0.013) in relation with severe toxicity of chemotherapy induced nausea and vomiting in breast cancer patients treated with Adriamycin chemotherapy. The 3435 C>T polymorphism of ABCB1 gene with heterozygous C/T genotype showed significantly negative association (OR=0.37, 95% CI: 0.14-0.96; p=0.042) with peripheral neuropathy in patients treated primarily with paclitaxel thereafter Adriamycin. CONCLUSION: The findings obtained from this study revealed significant association of ABCB1 3435 C>T polymorphisms with non-hematological toxicity in response to adriamycin and paclitaxel based chemotherapy.

Single-nucleotide polymorphism in CYP1A1, CYP1B1, CYP2B6, CYP2C8, and CYP2C9 genes and their association with gastrointestinal cancer: A hospital-based case-control study.

BACKGROUND: Cytochrome P450 (CYP) comprises a group of phase-I metabolizing enzymes that are important in xenobiotics metabolism. Genetic polymorphism of CYPs has been comprehensively studied for their association with a range of diseases. In this study, we assessed single-nucleotide polymorphism (SNP) of CYP1A, CYP1B, CYP2B, and CYP2C and their role in gastrointestinal (GI) cancer susceptibility in the rural population of Maharashtra. MATERIALS AND METHODS: In this hospital-based case-control study, the association of polymorphism of CYP genes was studied by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The study subjects included 200 clinically confirmed GI cancer patients and equal number of healthy controls. Odds ratio (OR) with 95% confidence interval (CI) and P value were evaluated to find out the level of association, where P ≤ 0.005 was considered statistically significant. RESULTS: After the analysis of CYP1A1*2A (rs4646903), CYP1B1*3 (rs1059836), CYP2B6*5 (rs3211371), CYP2C8*2 (rs11572103), CYP2C9*2 (rs1799853), and CYP2C9*3 (rs1057910), we noticed that variant (T) allele of CYP2B6*5 possessed significantly elevated risk (OR = 4.43; 95% CI: 2.20-8.90; P < 0.0001) of GI cancer in studied population. The genotypic distribution of G/C heterozygote allele of CYP1B1*3 (OR = 0.19, 95% CI = 0.12-0.32; P < 0.0001) and homozygous variant C/C allele (OR = 0.24, 95% CI = 0.13-0.45; P < 0.0001) showed a negative association with the development of GI cancer. CONCLUSION: The findings from this study supported that polymorphism of CYP2B6*5gene may be involved in the development of GI cancer. However, other SNPs of CYP1A, CYP1B, and CYP2C genes did not signify the risk for GI cancer in the studied population of rural Maharashtra.

Superoxide Dismutase (rs2070424, rs4880, rs2536512) and Catalase (rs794316, rs1001179) SNPs and their Association with Breast Cancer Risk: Findings from a Hospital Based Case-Control Study.

BACKGROUND: The antioxidant enzymes are important cellular components involved in detoxification of reactive oxygen species (ROS) and protect cells from ROS induced oxidative damage. Single nucleotide polymorphisms (SNPs) of antioxidant enzyme coding genes such as superoxide dismutase (SOD) and catalase (CAT) may alter the enzyme activity which can influence susceptibility towards carcinogenesis.  Therefore, the present study was planned to investigate possible SNPs of SOD (SOD1 (Cu,Zn-SOD), SOD2(Mn-SOD), SOD3(EC-SOD) and CAT genes and their possible association with breast cancer risk in rural Indian women. METHODS: In this case-control study, the association of SOD and CAT gene polymorphism was studied by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The study was conducted among 400 clinically breast cancer patients and 400 healthy women in a population of South-Western Maharashtra. The logistic regression analysis was carried out to calculate Odds ratio (OR) with 95% confidence interval and p-value, where p ≤0.05 was considered as statistically significant. RESULTS: The results of analysis of genotype frequency distribution showed significant association of rs4880 SNP of Mn-SOD with BC risk at homozygous variant (CC/CC) genotype (OR 2.46; 95%CI, 1.61-3.75; p<0.0001) and corresponding frequency of variant (C) allele (OR 1.53; 95%CI, 1.25-1.86; p<0.0001). In CAT gene polymorphisms the variant (T/T) was increased significantly in BC cases as compared to controls (OR 3.45; 95%CI, 2.17-5.50; p<0.0001) along with its variant (T) allele (OR 2.01; 95%CI, 1.63-2.48; p<0.0001). CONCLUSIONS: The results implied that, C/C genotype of SOD2-1183T/C polymorphism and T/T genotype of CAT-262 C/T polymorphism may be associated with an increased breast cancer risk. However, SOD1-251 A/G and SOD3-172 G/A polymorphisms did not show any significant difference in variant homozygous genotypes of patients compared to controls.

Genetic Polymorphisms of XPC, XPD, XPG Genes and their Association with Radiotherapy Induced Toxicity among Head and Neck Cancer Patients: A Hospital Based Study from Maharashtra.

BACKGROUND: The present study was planned to investigate possible association of single nucleotide polymorphisms (SNPs) of nucleotide excision repair (NER) genes such as XPC, XPD, XPG with acute radiation induced toxicities such as skin reactions and oral mucositis in normal tissue from head and neck cancer (HNC) patients receiving radiotherapy.  Methods: Two hundred and fifty HNC patients receiving radiotherapy were enrolled in this study and the acute toxicity reactions and radiation response were recorded. Association of SNPs rs2228001 of XPC, rs238406, rs13181 of XPD and rs17655 of XPG gene with normal tissue reactions in the form of dermatitis and mucositis were studied by PCR-RFLP and direct DNA sequencing. RESULTS: The results of univariate analysis of SNPs of XPC, XPD and XPG showed that XPC polymorphism at codon 939 of exon 15 (A>C) was not associated with dermatitis (OR=0.30, 95% CI: 0.06-1.39; p=0.125), or oral mucositis (OR=1.14, 95% CI: 0.41-3.20; p=0.793). The XPD codon 156 of exon 6 (C>A) and codon 751 of exon-23 A>C) polymorphism showed no association with radiosensitivity in HNC patients (OR=1.50, 95% CI: 0.60-3.71; p=0.080) for dermatitis, (OR=1.54, 95% CI: 0.66-3.61; p=0.312) for oral mucositis. The 1104 Asp variant genotype or allele of XPG (OR=1.35 95% CI: 0.50-3.64; p=0.541) showed no association with degree of radiotherapy associated dermatitis or mucositis (OR=0.80, 95% CI: 0.32-2.03; p=0.648) in HNC patients. The variant C allele of 2920 A/C genotype of XPC gene at codon 939 of exon 15, found protective with developing skin reactions with grade >1 (OR=0.60, 95% CI: 0.36-0.97; p=0.039) in HNC patients treated with radiotherapy. CONCLUSION: The results obtained in this study concluded that the SNPs rs2228001of XPC, rs238406, rs13181 SNPs of XPD and rs17655 SNP of XPG are not associated with normal tissue toxicity in HNC patients treated with radiotherapy. Radiotherapy with high radiation dose was significantly associated with oral mucositis in response to radiotherapy.

Single Nucleotide Polymorphisms in DNA Repair Genes (XRCC1, XRCC2, XRCC3) and Their Association with Radiotherapy Toxicity among Head and Neck Cancer Patients:A Study from South-Western Maharashtra.

BACKGROUND: The genetic polymorphisms in DNA repair genes and their correlation with normal tissue toxicity in response to radiation therapy has not been consistently proven in many of the studies done in head and neck cancers (HNC). This study was intended to investigate the association of most common single nucleotide polymorphisms of DNA repair genes with acute radiation induced toxicities such as skin reactions and oral mucositis in normal tissue from HNC patients receiving radiotherapy from South-Western Maharashtra. METHODS: Two hundred HNC patients receiving radiotherapy were enrolled in this study and the radiation injuries in the form of skin reactions and oral mucositis were recorded. Three single nucleotide polymorphisms (SNPs) rs1799782, rs25489) rs25487 of XRCC1 gene, rs3218536in XRCC2 gene and rs861539 SNP of XRCC3 gene were studied by PCR-RFLP and direct DNA sequencing.  Results: The univariate analysis of SNPs of XRCC1, XRCC2 and XRCC3, the obtained results verified that XRCC1 polymorphism at 194Trp of exon 6 (OR=0.69, 95% CI: 0.28-1.71; p=0.433), codon 280 at exon 9 ((OR=1.05, 95% CI: 0.42-2.63; p=0.911) and codon 399 of at exon 10(OR=1.06, 95% CI: 0.52-2.15; p=0.867) and XRCC2 polymorphism at codon 188 at exon 3 (OR=1.07, 95% CI: 0.46-2.47; p=0.866) and 241Met variant genotype of XRCC3 (OR=2.63 95% CI: 0.42-16.30; p=0.298) showed no association with degree of radiotherapy associated dermatitis or mucositis in HNC patients. CONCLUSION: The findings from this study postulated that none of rs1799782, rs25489, rs25487 SNPs of XRCC1, rs3218536 SNP of XRCC2 nor rs861539 SNP of XRCC3 were associated with increased toxicity of radiotherapy in HNC patients of south-western Maharashtra. 
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Impact of Interaction between Single Nucleotide Polymorphism of XRCC1, XRCC2, XRCC3 with Tumor Suppressor Tp53 Gene Increases Risk of Breast Cancer: A Hospital Based Case-Control Study.

BACKGROUND: At present very little information is available on combined effects of DNA repair genes with tumor suppressor gene polymorphisms and their association with cancer susceptibility. No such association studies have been carried out with breast cancer or any other cancer from India. Present study was conducted to study the combined effects of SNPs of XRCC1, XRCC2, XRCC3 with Arg72Pro and Arg249Ser SNPs of TP53 gene in risk of BC in rural parts of India. METHODS: The polymorphisms of Arg194Trp, Arg280His, Arg399Gln of XRCC1, Arg188His of XRCC2 and Thr241Met of  XRCC3 with Arg72Pro and Arg249Ser of TP53 gene polymorphisms was studied by polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) method. The association among the polymorphisms with breast cancer risk was studied by Odds ratio within 95% confidence interval and SNP-SNP interaction were confirmed by logistic regression analysis. RESULTS: The results of genotype frequency distribution of XRCC1, XRCC2, XRCC3 genotypes showed positive association between XRCC1 Arg280His polymorphism and BC risk (OR=4.54; 95% CI: 3.36- 6.15; p<0.0001).  Also the heterozygous genotypes Arg188His of XRCC2 (OR=1.58; 95% CI: 1.13- 2.21; p=0.007) and Thr241Met genotype of XRCC3 (OR=2.13; 95% CI: 1.44- 3.13; p=0.0001) were associated with BC risk. The combination of heterozygous Arg280His genotype of XRCC1 along with Arg72Pro genotype of TP53 increased the risk of BC (OR=4.53; 95% CI: 2.85-7.20); p<0.0001). Similarly,  the combined effect of heterozygous Arg/His genotype of XRCC1 with heterozygous Arg/Ser genotype of TP53 at codon 249 showed significant association with increased BC risk (OR=5.08; 95% CI: 2.86-9.04); p<0.0001). CONCLUSION: The findings derived from our study concluded that the heterozygous variant Arg280His genotype of XRCC1 and Thr241Met polymorphism of XRCC3 in combination with heterozygous arginine72proline genotype and heterozygous Arg249Ser polymorphism of TP53 showed significant association with breast cancer risk in Maharashtrian women.

Green synthesis of silver nanoparticles using Morinda citrifolia Linn leaf extract and its antioxidant, antibacterial and anticancer potential.

INTRODUCTION: Nanomedicine has emerged as a revolutionary regimen for moderating communicable as well as non-communicable diseases. PURPOSE: This study demonstrated the phytosynthesis of silver nanoparticles using M. citrifolia leaf extract (MC-AgNPs) and their in vitro antioxidant, antibacterial and anticancer potential. METHODS: The Biosynthesis of MC-AgNPs was studied by spectroscopy and characterized by SEM, TEM, XRD and FTIR analysis. The antibacterial activity was checked by minimum inhibition concentration assay. The HeLa and MCF-7 cancer cell lines were used to explore the cytotoxicity and genotoxicity activity of biogenic MC-AgNPs. RESULTS: The free radical scavenging potential of MC-AgNPs was studied by in vitro DPPH and ABTS assays, which confirmed significant radical scavenging activity in a dose-dependent manner with IC50 of 17.70 ± 0.36 µg/mL for DPPH and 13.37 ± 3.15 µg/mL for ABTS radicals. The bactericidal effects of MC-AgNPs confirmed by MIC showed 0.1 mg/mL concentration of MC-AgNPs with greater sensitivity for E.coli (93.33 ± 0.89), followed by K. pneumoniae (90.99 ± 0.57), S. aureus (87.26 ± 2.80) and P. aeruginosa strains (44.68 ± 0.73). The cytotoxicity results depicted strong dose and time-dependent toxicity of biogenic MC-AgNPs against cancer cell lines fifty percent inhibitory concentration MC-AgNPs against HeLa cells were 13.56 ± 1.22 µg/mL after 24h and 5.57 ± 0.12 µg/mL after 48 h exposure, likewise 16.86 ± 0.88 µg/mL and 11.60 ± 0.97 µg/mL respectively for MCF-7 cells. CONCLUSIONS: The present study revealed the green synthesis of silver nanoparticles using M. citrifolia and their significant antioxidant, antibacterial and anticancer activities.

Diabetic Foot Ulcer in India: Aetiological Trends and Bacterial Diversity.

Diabetes is one of the most prevalent epidemic metabolic disorders, responsible for a significant amount of physical, psychological and economic loss in human society. Diabetic foot ulcer (DFU) is one of the extreme pathophysiological consequences of diabetes. Bacterial infection is the most important cause of chronic DFU. Bacterial species or their biofilms show multidrug resistance, which complicates DFU and consequently leads to amputation of the infected part. Since the Indian population comprises diverse ethnic and cultural groups, this could influence the aetiology of diabetic foot infections and bacterial diversity. We reviewed 56 articles published from 2005 to 2022 on the microbiology of DFU and extracted the data on study location, number of patients analysed in the study, pathophysiological complications, age of the patients, sex of the patient, type of bacteria, type of infection (mono or polymicrobial), predominant bacteria (Gram-positive or Gram-negative), predominant isolates and multiple drug resistance (tested or not). We analysed data and described aetiological trends in diabetic foot infections and bacterial diversity. The study revealed that Gram-negative bacteria are predominant as compared to Gram-positive bacteria in individuals with diabetes with DFU in India. Escherichia coli, Pseudomonas aeruginosa, Klebsiella sp. and Proteus sp. were the most predominant Gram-negative bacteria, while Staphylococcus aureus and Enterococcus sp. were the major Gram-positive bacteria in DFU. We discuss bacterial infections in DFU in the context of bacterial diversity, sampling methods, demography and aetiology.

Comparative Study of Immunohistochemical Expression of Moesin and FLOT 1 in OSCC and Their Correlation with Histopathological Prognostic Factors.

AIM: To study immunohistochemical (IHC) expression patterns of Moesin and FLOT 1 in oral squamous cell carcinoma (OSCC) and to correlate it with histopathological prognostic factors. MATERIALS AND METHODS: A cross-sectional study design was conducted on histopathologically diagnosed cases of OSCC. The inclusion criteria were carcinoma of buccal mucosa, tongue, alveolar mucosa, palate, gingiva, the floor of the mouth, retromolar area, and soft palate. The exclusion criteria included cases of squamous cell carcinoma from sites other than the oral cavity, potentially malignant disorders (PMDs), and any pseudomalignancies of the head and neck. Tissue sections were subjected to IHC staining for Moesin and FLOT 1 and the results were subjected to statistical analysis. RESULTS: Moesin showed strong positivity and was significantly associated with the histopathological variables such as lymph nodes and the worst pattern of invasion, whereas FLOT 1 was not associated with any clinical, histopathological, or demographical variable in this study. CONCLUSION: Cytoplasmic detection of Moesin (35.19%) was higher than FLOT 1 (15.74%). There was no statistically significant relationship between the grade of the lesion and Moesin and FLOT 1. CLINICAL SIGNIFICANCE: New emerging prognostic biomarkers can aid to assess the rate of malignant transformation (epigenetic and molecular changes), thereby resulting in early prophylactic conciliation of the disease progression in OSCC. There is an urgent need for introducing these as an interventional therapy for effectively addressing OSCC at an early stage, thus preventing it from further proceeding to the advanced severe stage.

To evaluate an association between prognostic significance of Moesin with histopathological grading of oral squamous cell carcinoma: A systematic review.

Purpose: The aim of the present Aetiology/Risk type and Prognostic type of systematic review is to evaluate the value of Moesin as a biomarker of invasiveness in Oral Squamous Cell Carcinoma patients and to review/assess the available evidence regarding the prospective prognostic association between Moesin and histopathological grading of OSCC to enhance the quality of life and survival rate of oral cancer patients. Method: A systematic wide-range literature search was performed by authors (BS, KS, and DK) till October 2022 using both, electronic search media and manual search by hand, searching appropriate journals as per the focussed guiding question and inclusion/exclusion criteria. Major databases such as Scopus, EMBASE, Web of Science, Cochrane central register for controlled trials, PubMed & Google Scholar were conducted by two calibrated reviewers independently to gauge the association between the prognostic significance of Moesin with histopathological grading of oral squamous cell carcinoma. As this study is based on tissue samples of oral squamous cell carcinoma patients, all the selected studies were mostly, cross-sectional studies, and retrospective in nature. The studies were integrated with this review to gauge the association between the prognostic significance of Moesin with histopathological grading of oral squamous cell carcinoma (OSCC). The review included a total of 7 studies with tissue samples of 645 cases. The prime outcome was to assess the immunoexpression of Moesin among the different histopathological grades i.e well-differentiated SCC, moderately differentiated SCC, and poorly differentiated SCC and the subordinate outcome was to consider the extent of strong immunoexpression characteristics (cytoplasmic, membranous and mixed type) in different grades of OSCC as well as to correlate with morbidity, mortality, and/or 5 years or 10 years survival rate. Results: The results were analyzed and presented narratively using the Critical Appraisal Tools developed by the University Of Oxford; Risk of Bias - Cochrane Risk of Bias tool - RoB 2.0, and GRADE-pro (Grading of Recommendations, Assessment, Development, and Evaluations) which rates the features of the evidence as high, moderate, low and very low. The risk of mortality expressed in terms of Hazard ratio has been elicited as a 1.37 times higher rate of mortality in the advanced histopathological stages of the OSCC cases. As the sample size of this review was insignificant, therefore, the authors have incorporated hazard ratios of some other studies of carcinomas in diverse sites in the body to give a flavor of prognostic outcomes of Moesin. It was observed that Moesin expression in Breast cancer and UADT carcinomas have a higher mortality rate as compared to OSCC and lung carcinoma cases and this decree strengthens our conviction that Moesin expression in the cytoplasm of advanced histopathological stages of cancer can be assumed as a sign of poor prognosis in all carcinomas including OSCC patients. Conclusion: A sample of seven studies is inadequate as definite evidence for claiming that Moesin is a strong biomarker of invasiveness in OSCC cases and more clinical trials need to be conducted on the prognostic efficacy of Moesin expression in the various histopathological grades of OSCC cases.

TP53 (rs1042522, rs28934571) and TP21 (rs1801270, rs1059234) Polymorphisms and Risk of Breast Cancer among Rural Women of Maharashtra: Findings from a Hospital Based Case- Control Study.

BACKGROUND: Various studies all around the world depicted the relationship of polymorphisms in tumor suppressor genes with risk of various cancers, but there are unambiguous conclusions on this association. A hospital based case-control study was designed to review the association of polymorphism of tumor suppressor genes p21 and p53 with breast cancer risk in women residing in rural Maharashtra. METHODS: Two single nucleotide polymorphisms (SNPs) a C>A transversion (Ser>Arg) at codon 31 of exon 2 (rs1801270), C>T transition occurring 20bp upstream from stop codon of exon 3 (rs1059234) in p21 gene and G>C (Arg>Pro) transition at codon 72 of exon 4 (rs1042522), G>T (Arg>Ser) transition at codon 249 in exon 7 (rs28934571) in p53 gene were studied. To precise the quantitative assessment, we enrolled 800 subjects sorted into 400 clinically confirmed breast cancer patients and 400 healthy women from a tertiary care hospital (Krishna Hospital and Medical Research Centre) of south-western Maharashtra. The genetic polymorphisms in p21 and p53 genes was studied by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method using blood genomic DNA isolated from breast cancer patients and controls.  The level of association of polymorphisms was assessed using Odds ratio (OR) with 95% confidence interval and p-value identified using logistic regression model. RESULTS: After the analysis of SNPs (rs1801270, rs1059234) of p21 and (rs1042522, rs28934571) in p53 gene our analysis suggested that heterozygote Ser/Arg genotype with OR=0.66; 95% CI: 0.47- 0.91; p=0.0003 and homozygote variant Arg/Arg genotype with OR=0.23; 95% CI: 0.13- 0.40; p<0.0001of rs1801270 of p21 was negatively associated with risk of breast cancer in studied population. CONCLUSION: The findings from this study supported that rs1801270 SNP of p21 was inversely associated with breast cancer risk in the studied rural women population.

Biogenic Nanoparticles: Synthesis, Characterization, and Biological Potential of Gold Nanoparticles Synthesized using Lasiosiphon eriocephalus Decne Plant Extract.

INTRODUCTION: Recent advancements in biomedicine have revolutionized nanomedicine as a therapeutic moderator in the management of both infectious and noninfectious diseases. PURPOSE: In the current study we demonstrated biosynthesis of gold nanoparticles using aqueous leaf extract of Lasiosiphon eriocephalus as a capping and reducing agent and evaluation of their antioxidant, antibacterial, and anticancer properties. METHODS: The biosynthesized LE-AuNPs were characterized by UV-Vis spectrophotometry, SEM, TEM, XRD, FTIR, DLS, and Zeta potential analysis. The antibacterial activity was checked by a minimum inhibitory concentration assay. The anticancer potential of biogenic LE-AuNPs was checked by cytotoxicity and genotoxicity assay against HeLa and HCT-15 cells. RESULTS: The characteristic surface plasmon resonance peak of the colloidal solution at 538 nm by UV-Vis spectrum confirmed the formation of LE-AuNPs in the solution. The SEM, TEM, and XRD revealed 20-60 sized hexagonal and crystalline LE-AuNPs. The LE-AuNPs displayed significant inhibition potential against DPPH and ABTS radicals in vitro. The LE-AuNPs demonstrated significant antibacterial potential. The results of cytotoxicity interpreted that biogenic gold nanoparticles exhibited strong dose and time-dependent cytotoxicity effect against selected cancer cell lines where IC50 of LE-AuNPs required to inhibit the growth of HeLa cells after 24 h and 48 h exposure were 5.65± 0.69 µg/mL and 4.37±0.23 µg/mL respectively and that of HCT- 15 cells was 6.46 ± 0.69 µg/mL and 5.27 ± 0.34 µg/mL, 24h and 48h post-exposure respectively. CONCLUSIONS: Findings from this study revealed that gold nanoparticles synthesized using L. eriocephalus, showed remarkable antioxidant, antimicrobial, and extensive cytotoxicity and genotoxicity activities.