Prevalence of sleep disorders in patients with advanced cancer: a cross-sectional study.

Background: Patients with advanced cancer are more susceptible to develop sleep disorders like insomnia, restlessness, hypersomnolence, and sleep apnea due to a series of stressful events and side effects of chemotherapeutic agents. Poor sleep quality is associated with bad cancer outcomes and substandard quality of life. The authors assessed the prevalence of sleep disorders among advanced cancer patients in a tertiary center in Nepal. Methods: Patients with stage three and four solid malignancies were enrolled from February 2023 to July 2023 to assess their sleep status. The data were collected using the Pittsburgh Sleep Quality Index (PSQI) questionnaire, analyzed using the Statistical Package for the Social Sciences (SPSS) version 27, and subgroup exploration was done to assess the relationship of poor sleep quality with gender, marital status, malignancy type, and treatment received. An ethical clearance was obtained from the Institutional Review Committee (IRC). Results: The authors evaluated data from 357 patients in the study. Of them, 58.3% were female and 41.7% were male. The mean age of the patients was 51.1 years. Among total cancer patients, 56% had significant sleep disorders. A significant association was observed between the quality of sleep and gender, type of malignancy, and treatment methods (p value <0.05). A majority of the patients demonstrated increased sleep latency, struggling to fall asleep swiftly. Conclusions: More than half of the patients had poor sleep, which has an adverse impact on the prognosis of the disease and quality of life of cancer patients. Therefore, this aspect of cancer management requires special consideration for better quality of life and appropriate end-of-life care.

Differential TLR-ERK1/2 activity promotes viral ssRNA and dsRNA mimic-induced dysregulated immunity in macrophages.

RNA virus induced excessive inflammation and impaired antiviral interferon (IFN-I) responses are associated with severe disease. This innate immune response, also referred to as 'dysregulated immunity,' is caused by viral single-stranded RNA (ssRNA) and double-stranded-RNA (dsRNA) mediated exuberant inflammation and viral protein-induced IFN antagonism. However, key host factors and the underlying mechanism driving viral RNA-mediated dysregulated immunity are poorly defined. Here, using viral ssRNA and dsRNA mimics, which activate toll-like receptor 7 (TLR7) and TLR3, respectively, we evaluated the role of viral RNAs in causing dysregulated immunity. We show that murine bone marrow-derived macrophages (BMDMs) stimulated with TLR3 and TLR7 agonists induce differential inflammatory and antiviral cytokine response. TLR7 activation triggered a robust inflammatory cytokine/chemokine induction compared to TLR3 activation, whereas TLR3 stimulation induced significantly increased IFN/IFN stimulated gene (ISG) response relative to TLR7 activation. To define the mechanistic basis for dysregulated immunity, we examined cell-surface and endosomal TLR levels and downstream mitogen-activated protein kinase (MAPK) and nuclear factor kappa B (NF-kB) activation. We identified a significantly higher cell-surface and endosomal TLR7 expression compared to TLR3, which further correlated with early and robust MAPK (pERK1/2 and p-P38) and NF-kB activation in TLR7-stimulated macrophages. Furthermore, blocking EKR1/2, p38, and NF-kB activity reduced TLR3/7-induced inflammatory cytokine/chemokine levels, whereas only ERK1/2 inhibition enhanced viral RNA-mimic-induced IFN/ISG responses. Collectively, our results illustrate that high cell surface and endosomal TLR7 expression and robust ERK1/2 activation drive viral ssRNA mimic-induced excessive inflammatory and reduced IFN/ISG responses, and blocking ERK1/2 activity would mitigate viral-RNA/TLR-induced dysregulated immunity.

Breast Cancer among Cancer Patients Visiting the Department of Internal Medicine of a Tertiary Centre.

Introduction: Breast cancer is one of the most common cancers worldwide both in terms of incidence and mortality. Its incidence has been on an increasing trend in developing nations including Nepal, however, there is very limited evidence of its demographic profile in our setting. This study aimed to find out the prevalence of breast cancer among cancer patients visiting the Department of Internal Medicine of a tertiary centre. Methods: A descriptive cross-sectional study was conducted among cancer patients visiting the Department of Internal Medicine of a tertiary care centre using retrospectively collected data from 1 August 2022 to 30 July 2023 after obtaining ethical approval from the Institutional Review Committee. Histopathologically confirmed cancer patients were included and those with incomplete and duplicated data were excluded. A convenience sampling method was used. The point estimate was calculated at a 95% Confidence Interval. Results: Among 2067 cancer patients, the prevalence of breast cancer was 102 (4.93%) (4.00-5.87, 95% Confidence Interval). The mean age was 50.51±2.08 years. The most commonly affected age group was 40 to 60 years constituting 61 (59.80%) patients. Histologically, invasive breast cancer of no special type was the most common and was found in 91 (89.22%) patients. Conclusions: The prevalence of breast cancer was similar to other studies done in similar settings. Keywords: breast cancer; cross-sectional study; demography; malignancy; prevalence.

Mutation of a highly conserved isoleucine residue in loop 2 of several 𝛽-coronavirus macrodomains indicates that enhanced ADP-ribose binding is detrimental to infection.

All coronaviruses (CoVs) encode for a conserved macrodomain (Mac1) located in nonstructural protein 3 (nsp3). Mac1 is an ADP-ribosylhydrolase that binds and hydrolyzes mono-ADP-ribose from target proteins. Previous work has shown that Mac1 is important for virus replication and pathogenesis. Within Mac1, there are several regions that are highly conserved across CoVs, including the GIF (glycine-isoleucine-phenylalanine) motif. To determine how the biochemical activities of these residues impact CoV replication, the isoleucine and the phenylalanine residues were mutated to alanine (I-A/F-A) in both recombinant Mac1 proteins and recombinant CoVs, including murine hepatitis virus (MHV), Middle East respiratory syndrome coronavirus (MERS-CoV), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The F-A mutant proteins had ADP-ribose binding and/or hydrolysis defects that led to attenuated replication and pathogenesis in cell culture and mice. In contrast, the I-A mutations had normal enzyme activity and enhanced ADP-ribose binding. Despite increased ADP-ribose binding, I-A mutant MERS-CoV and SARS-CoV-2 were highly attenuated in both cell culture and mice, indicating that this isoleucine residue acts as a gate that controls ADP-ribose binding for efficient virus replication. These results highlight the function of this highly conserved residue and provide unique insight into how macrodomains control ADP-ribose binding and hydrolysis to promote viral replication.

Toll-like receptor 7 (TLR7)-mediated antiviral response protects mice from lethal SARS-CoV-2 infection.

SARS-CoV-2-induced impaired antiviral and excessive inflammatory responses cause fatal pneumonia. However, the key pattern recognition receptors that elicit effective antiviral and lethal inflammatory responses in-vivo are not well defined. CoVs possess single-stranded RNA (ssRNA) genome that is abundantly produced during infection and stimulates both antiviral interferon (IFN) and inflammatory cytokine/ chemokine responses. Therefore, in this study, using wild-type control and TLR7 deficient BALB/c mice infected with a mouse-adapted SARS-COV-2 (MA-CoV-2), we evaluated the role of TLR7 signaling in MA-CoV-2-induced antiviral and inflammatory responses and disease outcome. We show that TLR7-deficient mice are more susceptible to MA-CoV-2 infection as compared to infected control mice. Further evaluation of MA-CoV-2 infected lungs showed significantly reduced mRNA levels of antiviral type I (IFNα/ß) and type III (IFNλ) IFNs, IFN stimulated genes (ISGs, ISG15 and CXCL10), and several pro-inflammatory cytokines/chemokines in TLR7 deficient compared to control mice. Reduced lung IFN/ISG levels and increased morbidity/mortality in TLR7 deficient mice correlated with high lung viral titer. Detailed examination of total cells from MA-CoV-2 infected lungs showed high neutrophil count in TLR7 deficient mice compared to control mice. Additionally, blocking TLR7 activity post-MA-CoV-2 infection using a specific inhibitor also enhanced disease severity. In summary, our results conclusively establish that TLR7 signaling is protective during SARS-CoV-2 infection, and despite robust inflammatory response, TLR7-mediated IFN/ISG responses likely protect the host from lethal disease. Given similar outcomes in control and TLR7 deficient humans and mice, these results show that MA-CoV-2 infected mice serve as excellent model to study COVID-19.