Simian Immunodeficiency Virus Infection Mediated Changes in Jejunum and Peripheral SARS-CoV-2 Receptor ACE2 and Associated Proteins or Genes in Rhesus Macaques

Angiotensin converting enzyme-2 (ACE2) and associated proteins play a pivotal role in various physiological and pathological events, such as immune activation, inflammation, gut barrier maintenance, intestinal stem cell proliferation, and apoptosis. Although many of these clinical events are quite significant in SIV/HIV infection, expression profiling of these proteins has not been well reported. Considering the different pathological consequences in the gut after HIV infection, we hypothesized that the expression of ACE2 and associated proteins of the Renin-angiotensin system (RAS) could be compromised after SIV/HIV infection. We quantified the gene expression of ACE2 as well as AGTR1/2, ADAM17, and TMPRSS2, and compared between SIV infected and uninfected rhesus macaques (Macaca mulatta; hereafter abbreviated RMs). The gene expression analysis revealed significant downregulation of ACE2 and upregulation of AGTR2 and inflammatory cytokine IL-6 in the gut of infected RMs. Protein expression profiling also revealed significant upregulation of AGTR2 after infection. The expression of ACE2 in protein level was also decreased, but not significantly, after infection. To understand the entirety of the process in newly regenerated epithelial cells, a global transcriptomic study of enteroids raised from intestinal stem cells was performed. Interestingly, most of the genes associated with the RAS, such as DPP4, MME, ANPEP, ACE2, ENPEP, were found to be downregulated in SIV infection. HNFA1 was found to be a key regulator of ACE2 and related protein expression. Jejunum CD4+ T cell depletion and increased IL-6 mRNA, MCP-1 and AGTR2 expression may signal inflammation, monocyte/macrophage accumulation and epithelial apoptosis in accelerating SIV pathogenesis. Overall, the findings in the study suggested a possible impact of SIV/HIV infection on expression of ACE2 and RAS-associated proteins resulting in the loss of gut homeostasis. In the context of the current COVID-19 pandemic, the outcome of SARS-CoV-2 and HIV co-infection remains uncertain and needs further investigation as the significance profile of ACE2, a viral entry receptor for SARS-CoV-2, and its expression in mRNA and protein varied in the current study. There is a concern of aggravated SARS-CoV-2 outcomes due to possible serious pathological events in the gut resulting from compromised expression of RAS- associated proteins in SIV/HIV infection

An evaluation of patient satisfaction in outpatient departments of selected eye care hospitals in the Hyderabad City, Telangana State

The degree of agreement between a patient's expectations of optimal treatment and their evaluation of the actual care they got has been defined as the level of patient satisfaction. Medical service quality and cost-effectiveness, doctor conduct and attitude, the cost of services, hospital infrastructural facilities, physical comfort and psychological care, as well as empathy for clients' testimonies all go towards a patient's satisfaction with their healthcare facility experience. To determine how satisfied patients are with the out-patient department (OPD) services at five different eye care facilities in Hyderabad, Telangana, and the variables influencing patient satisfaction. To conduct the study, the out-patient department of LV Prasad Eye Institute: Banjara Hills, Vasavi Eye Care & Diabetic Center: Chandanagar, Aravind Eye Hospital & Institute: Mehdipatnam, Sri Madhavi Eye Care Hospital: Habsiguda, Dr. Agarwal Eye Hospital: Panjagutta in Hyderabad city, Telangana state was visited by researchers from 01.04.2021 to 30.06.2021. To choose patients for the out-patient section of the hospital, a systematic random technique was adopted

Antibiotic Susceptibility of Bartonella Grown in Different Culture Conditions

Bartonellosis is caused by a Gram-negative intracellular bacterium with a zoonotic transmission. The disease, caused by any of several genospecies of Bartonella can range from a benign, self-limited condition to a highly morbid and life-threatening illness. The current standard of care antibiotics are generally effective in acute infection; these include azithromycin or erythromycin, doxycycline, gentamicin, rifampin, and ciprofloxacin. However, treatment of chronic infection remains problematic. We tested six different antibiotics for their ability to stop the growth of Bartonella sp. in the standard insect media and in an enrichment media. All antibiotics (ceftriaxone, doxycycline, gentamycin, azithromycin, ampicillin, and azlocillin) had minimum inhibitory concentrations (MICs) below 0.5 µg/mL in the BAPGM enrichment media but were ineffective at inhibiting growth when the standard insect media was used. Azlocillin was the most potent, with a MIC of 0.01 µg/mL. When Bartonella was tested under intracellular growth conditions, none of the antibiotics were efficacious singly. However, growth inhibition was observed when azlocillin and azithromycin were combined. These studies illustrate the impact of growth medium and intracellular environment on antibiotic susceptibility testing and indicate that azlocillin combined with azithromycin may be an effective drug combination for the treatment of Bartonellosis

Empirical Investigation on Measurement of Game Immersion using Real World Dissociation Factor

Context: Games involve people to a large extent where they relate themselves with the game characters; this is commonly known as game immersion. Generally, some players play games for enjoyment, some for stress relaxation and so on.Game immersion is usually used to describe the degree of involvement with a game. When people play games, they don’t necessarily realize that they have been dissociated with the surrounding world. Real world dissociation (RWD) can be defined as the situation where a player is less aware of the surroundings outside the game than about what is happening in the game itself. The RWD factor has been expected to measure the losing track of time, lack of awareness of surroundings and mental transportation. Objectives: In this thesis, we measure and compare the difference in game immersion between experienced and inexperienced players using RWD factor. In addition, the study involves exploring the significance of game immersion and various approaches used to measure it. Methods: In this study literature review has been carried out to explore the meaning of game immersion and further user studies in the form of an experiment has been conducted to measure game immersion between experienced and inexperienced gamers. The game immersion has been measured using the real world dissociation (RWD) factor. After the experiment has been conducted, a statistical technique has been carried out to measure the difference in game immersion among the two groups. Results:The empirical investigation on the measurement of game immersion has been done using RWD factor. The results state that the significance value is less than 0.05 and hence null hypothesis is rejected for both the games. The measurable difference has been calculated by using Cohen’s d effect size between experienced and inexperienced players. The Cohen’s d value between experienced players and inexperienced players for Dota 2 is 0.7423 and CS:GO is 0.8383. Conclusions: After analyzing the data and calculating the effect size, the overall results state that inexperienced group of players are more immersed than the experienced group of players when measured by RWD factor. Hence it can be concluded that irrespective of the game played, inexperienced players are more dissociated from the real world than the experienced players

Cargo trafficking from the trans-Golgi network towards the endosome

The trans‐Golgi network (TGN) is a major sorting, packing and delivering station of newly synthesised proteins and lipids to their final destination. These cargo molecules follow the secretory pathway, which is a vital part of cellular trafficking machinery in all eukaryotic cells. This secretory pathway is well conserved in all eukaryotes from low‐level eukaryotes, such as yeast, to higher level eukaryotes like mammals. The molecular mechanisms of protein sorting by adaptor proteins, membrane elongation and transport to the final destinations by motor proteins and the cytoskeleton, and membrane pinching‐off by scission proteins must be choreographically managed for efficient cargo delivery, and the understanding of these detailed processes is not yet completed. Functionally, defects in these mechanisms are associated with the pathology of prominent diseases such as acute myeloid leukaemia, Charcot-Marie-Tooth disease, I‐cell disease and Wiskott-Aldrich syndrome. The present review points out the recent advances in our knowledge of the molecular mechanisms involved in the transportation of the cargo from the TGN towards the endosome

Automated design of protein-binding riboswitches for sensing human biomarkers in a cell-free expression system

Abstract Cell-free genetically encoded biosensors have been developed to detect small molecules and nucleic acids, but they have yet to be reliably engineered to detect proteins. Here we develop an automated platform to convert protein-binding RNA aptamers into riboswitch sensors that operate within low-cost cell-free assays. We demonstrate the platform by engineering 35 protein-sensing riboswitches for human monomeric C-reactive protein, human interleukin-32γ, and phage MS2 coat protein. The riboswitch sensors regulate output expression levels by up to 16-fold with input protein concentrations within the human serum range. We identify two distinct mechanisms governing riboswitch-mediated regulation of translation rates and leverage computational analysis to refine the protein-binding aptamer regions, improving design accuracy. Overall, we expand the cell-free sensor toolbox and demonstrate how computational design is used to develop protein-sensing riboswitches with future applications as low-cost medical diagnostics

Direct Capture and Early Detection of Lyme Disease Spirochete in Skin with a Microneedle Patch

Borrelia burgdorferi sensu lato family of spirochetes causes Lyme disease (LD) in animals and humans. As geographic territory of ticks expands across the globe, surveillance measures are needed to measure transmission rates and provide early risk testing of suspected bites. The current standard testing of LD uses an indirect two-step serological assay that detects host immune reactivity. Early detection remains a challenge because the host antibody response develops several weeks after infection. A microneedle (MN) device was developed to sample interstitial fluid (ISF) and capture spirochetes directly from skin. After sampling, the MN patch is easily dissolved in water or TE buffer, and the presence of spirochete DNA is detected by PCR. Performance was tested by spiking porcine ear skin with inactivated Borrelia burgdorferi, which had an approximate recovery of 80% of spirochetes. With further development, this simple direct PCR method could be a transformative approach for early detection of the causative agent of Lyme disease and enable rapid treatment to patients when infection is early, and numbers of systemic spirochetes are low

A comparison of Bartonella henselae infection in immunocompetent and immunocompromised mice.

Bartonellosis refers to disease caused by the Bartonella genus of bacteria. The breadth of disease manifestations associated with Bartonella is currently expanding and includes regional lymphadenopathy, rheumatic, ocular, and neurological disorders. The dearth of knowledge regarding diagnosis, treatment and pathogenesis of this disease can be partially attributed to the lack of a reliable small animal model for the disease. For this study, Bartonella henselae, the most common species associated with human disease, was injected into Swiss Webster (SW) mice. When the outcome indicated that productive infection did not occur, SCID/Beige (immune compromised) mice were inoculated. While SW mice may potentially harbor an acute infection, less than 10 days in length, the SCID/Beige model provided a sustained infection lasting up to 30-days. These data indicate that SCID/Beige mice can provide a model to study Bartonella infection, therapeutics, and vector dynamics in the future