Identification of a novel ER-NFĸB-driven stem-like cell population associated with relapse of ER+ breast tumors.

BACKGROUND: Up to 40% of patients with estrogen receptor-positive (ER+) breast cancer experience relapse. This can be attributed to breast cancer stem cells (BCSCs), which are known to be involved in therapy resistance, relapse, and metastasis. Therefore, there is an urgent need to identify genes/pathways that drive stem-like cell properties in ER+ breast tumors. METHODS: Using single-cell RNA sequencing and various bioinformatics approaches, we identified a unique stem-like population and established its clinical relevance. With follow-up studies, we validated our bioinformatics findings and confirmed the role of ER and NFĸB in the promotion of stem-like properties in breast cancer cell lines and patient-derived models. RESULTS: We identified a novel quiescent stem-like cell population that is driven by ER and NFĸB in multiple ER+ breast cancer models. Moreover, we found that a gene signature derived from this stem-like population is expressed in primary ER+ breast tumors, endocrine therapy-resistant and metastatic cell populations and predictive of poor patient outcome. CONCLUSIONS: These findings indicate a novel role for ER and NFĸB crosstalk in BCSCs biology and understanding the mechanism by which these pathways promote stem properties can be exploited to improve outcomes for ER+ breast cancer patients at risk of relapse.

Geriatrics fellowship during COVID-19: expanding impact, using innovation and maintaining balance.

The COVID-19 pandemic provided an opportunity for geriatricians, especially geriatrics fellows, to demonstrate leadership in a crisis that has significantly affected the 65 and older demographic. Given their expertise in care delivery to complex, multimorbid patients, as well as their ability to navigate different healthcare settings, geriatrics fellows became a valuable resource during the pandemic, particularly at one large, urban academic health system. Their training in patient-centered, value-based care helped determine the best course of action for patients not only in the hospital, but also in the community. Utilizing innovative strategies such as a newly developed Palliative Care Hotline (PATCH-24 line), telehealth, and community paramedicine, fellows delivered services to complex patients in community settings. In addition to providing direct patient care, geriatrics fellows also taught their skills to frontline physicians of other specialties. Strong support from the fellowship program's leadership, as well as an ongoing focus on clinician wellbeing and resilience, have been central factors in the success of geriatrics fellows during the COVID-19 crisis.

A Study of the Outcome of Critically Ill Elderly Patients in a Tertiary Care Hospital Using SOFA Score.

BACKGROUND: With a rapidly rising geriatric population, the magnitude of elderly patients requiring intensive care is a major cause of concern. Data on critically ill geriatric patients is scarce, especially in developing countries. AIM AND OBJECTIVE: The aim of the study is to identify the etiology, clinical profile, and outcome in elderly patients admitted to the intensive care unit (ICU) and to predict their survival using the sequential organ failure assessment (SOFA) score. MATERIALS AND METHODS: A prospective observational study was performed over a period of 18 months with analysis of 100 patients admitted to the ICU, above the age of 60 years, with multi-organ dysfunction. The outcome of discharge or death was studied using the SOFA score on admission, on day 2, and the delta SOFA score. RESULTS: In this study of 100 patients, 88% of patients were in the 60-70 years age-group. The number of male and female patients was equal. Seventy percent of patients had comorbidities, of which hypertension was most common. The two most common etiologies were acute febrile illness and pneumonia. The use of mechanical ventilation, inotropic support, and serum creatinine has a significant association with the outcome. The SOFA score at admission did not have a significant association, but the score at 48 hours and delta SOFA score co-related with the outcome of the patients. Sixty-four patients got discharged; thus, there was a survival rate of 64%. CONCLUSION: The SOFA score at 48 hours is the most sensitive predictor of outcome, followed closely by the delta SOFA score, as compared to the SOFA score on admission, for critically ill elderly patients. There is a significant association of use of mechanical ventilation, inotropic support, and serum creatinine with the outcome. HOW TO CITE THIS ARTICLE: Chopra S, Pednekar S, Karnik ND, Londhe C, Pandey D. A Study of the Outcome of Critically Ill Elderly Patients in a Tertiary Care Hospital Using SOFA Score. Indian J Crit Care Med 2021;25(6):655-659.

Potential of combination therapy of endolysin MR-10 and minocycline in treating MRSA induced systemic and localized burn wound infections in mice.

MRSA is the predominant pathogen responsible for fatal burn wound infection in patients. Antibiotic resistance and its ability to form biofilms on the surface of burn wounds limit the use of antibiotics to contain this pathogen. The results of present study have shown that single dose of combination therapy of endolysin MR-10 (50µg/s.c) and minocycline (50mg/kg/orally) resulted in 100% survival of group of mice with systemic MRSA infection. Maximum reduction in bacterial load in various organs was observed in the group that received combination therapy. In comparison to control, a significant reduction (p<0.01) of 4.82, 1.81, 1.51, 1.2 logs was observed in skin, blood, liver and spleen respectively, by 3rd day post infection. As a result of which, all organs became sterile thereby protecting mice from mortality. Histopathological analysis corroborated our findings showing no signs of inflammation and bacterial infection in the group that received combination therapy. Treatment of localized burn wound infection with combination therapy resulted in early resolution of infection followed by fast healing. The group that received combination therapy showed complete resolution of infection in less than 10days. Moreover, the skin samples obtained from animals treated with combination therapy showed no myeloperoxidase (MPO) activity on 10th day post treatment. In combination therapy group, ∼98% wound contraction was observed by 12th day followed by complete closure of wound within ∼14days. The histopathological analysis showed no signs of inflammation and infection. Collagen staining revealed early signs of re-epithelization of epidermis and signs of collagen regeneration in-group that received combination therapy. Hence, this study suggests that the combined therapy of endolysin MR-10 and minocycline is a better option in controlling burn wound infections.

Potential of sequential treatment with minocycline and S. aureus specific phage lysin in eradication of MRSA biofilms: an in vitro study.

Lysins are novel class of anti-infectives which are derived from bacteriophage. In the present study, the potential of previously characterised phage borne endolysin MR-10 in eradicating methicillin-resistant Staphylococcus aureus (MRSA) biofilm was evaluated. Scanning electron microscopic examination showed that both ica-positive and ica-negative MRSA formed equally potent mature biofilm. Different approaches were employed to eradicate the young as well as older biofilm formed by both types of MRSA strains. Our results showed a significant decrease (p < 0.01) in biofilm count on sequentially treating the MRSA biofilm with minocycline (4 µg/ml) for 3 h followed by treatment with endolysin MR-10. Since endolysin can act effectively irrespective of the metabolic status of the cells hence, they are capable of killing the rapidly growing cells (log phase cells) as well as non-dividing (stationary phase) cells. As a result they are effective in eradicating the younger and older biofilm. On staining the ica-positive MRSA biofilm with wheat germ agglutinin (WGA)-Alexa Flour 350, reduction in poly-intercellular adhesion (PIA) content was observed in comparison to control biofilm. In addition, a significant decrease (p < 0.01) in extracellular DNA (eDNA) content of ica-negative MRSA biofilm was also observed. Further, Live/Dead Baclight™ staining also showed presence of higher population of dead cells after treatment with minocycline and endolysin MR-10. Hence, our results showed that using minocycline sequentially with endolysin, MR-10 can effectively eradicate both young as well as older biofilm formed by ica-positive and ica-negative MRSA.

Antibiotic susceptibility of ica-positive and ica-negative MRSA in different phases of biofilm growth.

Methicillin-resistant Staphylococcus aureus (MRSA) has emerged as a clinically relevant pathogen because of its resistance to antibiotics and its ability to form potent biofilm. Both ica-negative as well as ica-positive MRSA strains are known to produce biofilm. In the present study, these strains were grown in biofilm mode and susceptibility of these to antibiotics was assessed. Our study suggests that antibiotic susceptibility of MRSA biofilm depends on the biochemical composition of its matrix. The biofilm matrix of ica-positive MRSA was mainly composed of poly-intercellular adhesion (PIA), whereas eDNA was a major constituent of ica-negative MRSA. The results showed that MRSA in planktonic growth was susceptible to clindamycin, vancomycin and minocycline. However, the MIC and MBC of vancomycin for the mature biofilm of ica-negative MRSA was 16 and 32 µg ml(-1), respectively. On the contrary, the MIC and MBC of vancomycin for ica-positive MRSA was >1024 µg ml(-1). The effect of vancomycin and minocycline on young and old biofilms was also determined. Vancomycin was quite effective in eradicating the young biofilm formed by ica-negative MRSA; however, it was completely ineffective on the biofilm of ica-positive MRSA. Minocycline at its highest clinical achievable concentration was found to be quite effective in eradicating the young biofilm formed by both the strains. The enzyme-linked immunosorbent assay (ELISA) results and dot blot assay suggest that the presence of ica locus influenced PIA production, which probably contributed towards the failure of vancomycin in eradicating the biofilm formed by ica-positive strain. However, none of the antibiotics used in this study was effective in eradicating the mature biofilms.