Search

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 observed that murine bone marrow-derived macrophages (BMDMs), when stimulated with TLR3 and TLR7 agonists, induced 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 significantly higher cell-surface and endosomal TLR7 levels compared to TLR3, which were associated with early and robust MAPK (p-ERK1/2, p-P38, and p-JNK) and NF-kB activation in TLR7-stimulated macrophages. Furthermore, blocking EKR1/2 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 response and blocking ERK1/2 activity would likely mitigate viral-RNA/TLR-induced dysregulated immunity. [ABSTRACT FROM AUTHOR]

The inhabitants of Nepal's hilly and Himalayan regions heavily depend on herbs and medicinal plants as a primary source of livelihood. In rural hilly and mountainous areas, the collection and trade of herbs remain the predominant source of income for a significant portion of the Nepalese population. Despite the economic importance of herb collection and trade, there has been limited systematic work on pest identification for exportable herbs in Nepal. This lack of study results in restricted options for effectively controlling diseases and insect pests affecting these herbs and their export. To address this gap, a comprehensive survey of major exportable herbs--Amla (Phyllanthus emblica), Pakhanved (Bergenia ciliata), and Nagbeli (Lycopodium clavatum)--was conducted from October 2021 to July 2022 in these herbs-producing major districts (Gorkha, Kaski, Dailekh, Salyan, Dhankuta, Dhading, Baglung, and Dolakha) in Nepal. The survey aimed to assess the distribution and occurrence of common diseases and pests in Nepal. The survey findings indicated that certain pathogens and pests were widespread, while others exhibited a more localized distribution. Specifically, the survey identified a total of 7 diseases, 8 insects, and 15 weeds affecting Amla, 4 diseases, 3 insects, and 3 weeds affecting Pakhanved, and 3 diseases, 5 insects, and 19 weeds affecting Nagbeli. These results offer valuable insights into the national status of pest issues related to exportable herbs in Nepal. These findings provide a localized perspective on the specific diseases and pests affecting the surveyed regions, offering valuable insights for targeted management and control strategies. [ABSTRACT FROM AUTHOR]

Introduction: The second most prevalent cancer among women in Nepal is breast cancer. The assessment of human epidermal growth factor receptor-2 (HER2), progesterone receptor (PR), and estrogen receptor (ER) status in patients with breast cancer is currently regarded as standard practice because of its prognostic and predictive value. This study aims to find the incidence of Hormone receptor and HER2 status in patients with breast cancer. Methods: This was a descriptive observational cross-sectional study that assessed the immunohistochemistry markers of different histological types of breast cancer patients who visited a tertiary center in Nepal. Retrospective data from August 1, 2022, to July 30, 2023, were gathered from computer records of all the patients diagnosed with breast cancer who visited the Tribhuvan University Teaching Hospital (TUTH). SPSS version 26.0 was used for statistical analysis. Results: A total of 102 patients with breast cancer were included. The mean age of the patients was 50.51±12.08 years. Only one patient was male. The analysis showed that 62 (60.78%) of total patients had ER/PR positive breast cancer and 40 (39.2%) were HER2 positive with a higher number among Janajati and the age group 41-60 years. Similarly, the incidence of triple-negative breast cancer was 21 (20.59%) of all patients. Conclusion: The majority of breast cancer patients in our study were ER/PR positive, followed by HER2-positive and triple-negative cases. A higher prevalence was observed among the Janajati ethnic group and those aged 41-60 years. [ABSTRACT FROM AUTHOR]

Veterinary hematology serves as an important screening procedure to assess general health conditions, diagnosis, and treatment of disease. This study aims to interpret and establish a set of hematology reference ranges for Asian Elephants managed by private and government facilities in Nepal. Blood samples from 50 elephants around Chitwan National Park, Sauraha were collected and hematological parameters such as total erythrocyte count and total leukocyte count were determined. The results show that the majority of hematological values were in line with the values previously published by different authors. The mean erythrocyte and leukocyte counts were reported as 3.32±0.93 x 10^6 cell/μL and 10448±335.49 cells/μL respectively. No sex-associated significant difference was observed in the case of total erythrocyte count, whereas total leukocyte counts varied significantly within sexes. Our findings revealed no significant difference in hematological parameters between government and privately owned elephants. The hematological values from our study can be used as reference values for assessing the health condition of elephants in Nepal. Further research work should be conducted to evaluate the factors affecting hematological parameters. [ABSTRACT FROM AUTHOR]

All coronaviruses (CoVs) encode for a conserved macrodomain (Mac1) located in non-structural protein 3. 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 glycine-isoleucine-phenylalanine motif. While we previously demonstrated the importance of the glycine residue for CoV replication and pathogenesis, the impact of the isoleucine and phenylalanine residues remains unknown. 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, 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 correlated with attenuated replication and pathogenesis of F-A mutant MERS-CoV and SARS-CoV-2 viruses in cell culture and mice. In contrast, the I-A mutant proteins had normal enzyme activity and enhanced ADP-ribose binding. Despite only demonstrating increased ADP-ribose binding, I-A mutant MERS-CoV and SARS-CoV-2 viruses 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 ADPribose binding and hydrolysis to promote viral replication. IMPORTANCE The conserved coronavirus (CoV) macrodomain (Mac1) counters the activity of host ADP-ribosyltransferases and is critical for CoV replication and pathogenesis. As such, Mac1 is a potential therapeutic target for CoV-induced disease. However, we lack a basic knowledge of how several residues in its ADP-ribose binding pocket contribute to its biochemical and virological functions. We engineered mutations into two highly conserved residues in the ADP-ribose binding pocket of Mac1, both as recombinant proteins and viruses for Middle East respiratory syndrome coronavirus (MERS-CoV) and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Interestingly, a Mac1 isoleucine-to-alanine mutant protein had enhanced ADP-ribose binding which proved to be detrimental for virus replication, indicating that this isoleucine controls ADP-ribose binding and is beneficial for virus replication and pathogenesis. These results provide unique insight into how macrodomains control ADP-ribose binding and will be critical for the development of novel inhibitors targeting Mac1 that could be used to treat CoV-induced disease. [ABSTRACT FROM AUTHOR]

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., Competing Interests: The authors have no competing interests.

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.

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.