Histopathologically Confirmed Pulmonary Mucormycosis as a Complication of COVID-19: a Case Report from Romania and Insight into Pathology.

COVID-19 has proven to be an independent risk factor for secondary infectious complications. Amongst them, mucormycosis has recently been noticed more frequently than in the past. Caused by molds belonging to the Mucorales order, this is a rare, but potentially fatal infection unless adequately treated. Ear, nose and throat involvement is prevalent with often expansion to the orbit, sinuses or brain. Pulmonary, cutaneous and gastrointestinal infections are also recognized. Classical risk factors for progression to angioinvasive disease include poorly controlled diabetes mellitus, defects in phagocytic function (prolonged neutropenia, glucocorticoid treatment), immunosuppressive therapy associated with transplantation, malignancy, elevated levels of free iron as well as iron chelators (deferoxamine). In addition, immune dysregulation rendered by COVID-19 itself may contribute or solely lead to invasive mold disease. The largest experience comes from India, which has dealt with a challenging epidemic of COVID-19-associated mucormycosis (CAM). To our knowledge, no previous studies have reported CAM in Romania. We therefore present a case of severe COVID-19 pneumonia initially complicated by bacterial superinfection and secondary sepsis at admission in an unvaccinated 61-year-old male who presented in our clinic with respiratory failure and digestive symptoms. Although improvement occurred rapidly following antiviral, empiric large spectrum Intraantibiotics and pathogenic medication, unfavorable clinical course ensued later on. Biological and imaging investigations were consistent with pulmonary superinfection in the form of multiple different-sized upper right field opacities, which eventually evolved to form cavities. Differential diagnosis was thoroughly performed. Since unable to sterilize the lung by means of medication alone, the patient underwent major thoracic surgery with removal of the entire right lung. Microscopic study of the damaged tissue was able to determine the presence of broad, aseptate hyphae which morphologically belong to Mucorales. A diagnosis of pulmonary mucormycosis was established and proper antifungal treatment was initiated, with full recovery of the patient.

G protein-coupled receptor interactions and modification of signalling involving the ghrelin receptor, GHSR1a.

The growth hormone secretagogue receptor 1a (GHSR1a) is intriguing because of its potential as a therapeutic target and its diverse molecular interactions. Initial studies of the receptor focused on the potential therapeutic ability for growth hormone (GH) release to reduce wasting in aging individuals, as well as food intake regulation for treatment of cachexia. Known roles of GHSR1a now extend to regulation of neurogenesis, learning and memory, gastrointestinal motility, glucose/lipid metabolism, the cardiovascular system, neuronal protection, motivational salience, and hedonic feeding. Ghrelin, the endogenous agonist of GHSR1a, is primarily located in the stomach and is absent from the central nervous system (CNS), including the spinal cord. However, ghrelin in the circulation does have access to a small number of CNS sites, including the arcuate nucleus, which is important in feeding control. At some sites, such as at somatotrophs, GHSR1a has high constitutive activity. Typically, ghrelin-dependent and constitutive GHSR1a activation occurs via Gαq/11 pathways. In vitro and in vivo data suggest that GHSR1a heterodimerises with multiple G protein-coupled receptors (GPCRs), including dopamine D1 and D2, serotonin 2C, orexin, oxytocin and melanocortin 3 receptors (MCR3), as well as the MCR3 accessory protein, MRAP2, providing possible mechanisms for its many physiological effects. In all cases, the receptor interaction changes downstream signalling and the responses to receptor agonists. This review discusses the signalling mechanisms of GHSR1a alone and in combination with other GPCRs, and explores the physiological consequences of GHSR1a coupling with other GPCRs.

New Waste-Based Composite Material for Construction Applications.

The global demand for fiber-based products is continuously increasing. The increased consumption and fast fashion current in the global clothing market generate a significant quantity of pre-and post-production waste that ends up in landfills and incinerators. The present study aims to obtain a new waste-based composite material panel for construction applications with improved mechanical properties that can replace traditional wood-based oriented strand boards (OSB). The new composite material is formed by using textile wastes as a reinforcement structure and a combination of bi-oriented polypropylene films (BOPP) waste, polypropylene non-woven materials (TNT) waste and virgin polypropylene fibers (PP) as a matrix. The mechanical properties of waste-based composite materials are modeled using the Taguchi method based on orthogonal arrays to maximize the composite characteristics' mechanical properties. Experimental data validated the theoretical results obtained.

Determination of Physicomechanical Characteristics of the Cement Mortar with Added Fiberglass Waste Treated with Hydrogen Plasma.

Solving the environmental problems and the economic aspects of the construction sector represent a global priority. The considerable quantities of raw materials and the energy consumed by this sector make it one of the most polluting economic activities. Fiberglass in various forms is widely used in the construction sector. In the manufacturing process and during the usage of fiberglass products, a significant amount of indestructible waste results, negatively impacting the environment. An innovative solution for utilizing this type of waste is the treatment with hydrogen plasma. This process results in two products: the first in the gaseous state used to obtain synthetic fuel and the second in solid-state, named slag. The composition of solid waste contains chemical compounds that can increase their strength if used as additives in mortars or concretes. This study presents the laboratory tests on mortars, in which a part of the cement amount was replaced with the solid component resulting from the plasma treatment of glass fiber waste. The results showed that replacing a part of the cement with these materials is a solution that minimizes the ecological footprint of the buildings.

Air Distribution in a Fully-Closed Higher Plant Growth Chamber Impacts Crop Performance of Hydroponically-Grown Lettuce.

The MELiSSA Pilot Plant (MPP) is testing in terrestrial conditions regenerative life support technologies for human exploration in Space. One of its components is a controlled Higher Plant Chamber (HPC) accommodating hydroponic plant cultures. It consists of a 9 m3 single closed growth chamber providing adequate environmental conditions for growing plants, enabling the production of food, water and oxygen for the crew. A critical aspect for a reliable HPC performance is to achieve homogeneous air distribution. The initial experiment carried out in the MPP with lettuce as salad crop, showed uneven plant growth throughout the HPC, which was attributed to inadequate air distribution due to non-homogeneous air velocity profile along the inlet-vents. After a detailed computational fluid dynamics (CFD) analysis, the heating, ventilation, and air conditioning subsystem of the HPC was upgraded and a new experiment was carried out in optimized air flow conditions. Nine-day seedlings of lettuce cultivar "Grand Rapids" were transplanted into the HPC and harvested at the end of the growing cycle, where shoot fresh weight, dry biomass, and shoot mineral composition were analyzed. During the experiment, the environmental control system performed remarkably well based on the biometric measurements as well as the mineral composition leading to a vast homogeneous growth. Overall, the results demonstrated the beneficial effect of an adequate air distribution system in HPCs and the effectiveness of CFD-analysis to design properly the gas distribution. The obtained results are of high relevance for life support systems in space involving plants growth.

Conformational Transitions and the Activation of Heterotrimeric G Proteins by G Protein-Coupled Receptors.

G protein-coupled receptors (GPCRs) are particularly attractive targets for therapeutic pharmaceuticals. This is because they are involved in almost all facets of physiology, in many pathophysiological processes, they are tractable due to their cell surface location, and can exhibit highly textured pharmacology. While the development of new drugs does not require the molecular details of the mechanism of activity for a particular target, there has been increasing interest in the GPCR field in these details. In part, this has come with the recognition that differential activity at a particular target might be a way in which to leverage drug activity, either through manipulation of efficacy or through differential coupling (signaling bias). To this end, the past few years have seen a number of publications that have specifically attempted to address one or more aspects of the molecular reaction pathway, leading to activation of heterotrimeric G proteins by GPCRs.

"Identification of Non-Tuberculous Mycobacterium by LPA (CM/AS) assay, HPLC and biochemical test: which is feasible for RNTCP?"

INTRODUCTION: Non-tuberculous mycobacteria (NTM) causing clinical disease have become increasingly common and more diverse. The development of fast, inexpensive, and reliable tests to identify nontuberculous mycobacteria is need of the hour especially under the Revised National TB Control Programme (RNTCP). The Aim of the study was to check the Diagnostic efficacy of the GenoType Mycobacterium CM/AS assay compared with HPLC and Biochemical Test for Identification of Non-Tuberculous Mycobacteria under the Revised Tuberculosis Control Programme. METHODS AND RESULT: It is a cross-sectional study, the suspected NTM culture isolates from the RNTCP accredited laboratories were sent to NRL for speciation and Identification. The culture positive isolates were subjected to Biochemical Identification Test, HPLC and LPA CM/AS. The LPA had 98.23% sensitivity, 50% specificity, 99.56% positive predictive value (PPV) and 20% Negative predictive value (NPV) when compared to HPLC considering Biochemical test as Gold reference standard. The comparison of HPLC and LPA for identification of each species using Mc Nemers Chi square test shown no significant difference between these tests. CONCLUSION: Considering Cost, Time and ease of performing the techniques, we recommend first do the basic biochemical test to rule out MTBC from NTM. Then do HPLC and further if results are unclear do LPA CM/AS kit for species confirmation. SIGNIFICANCE AND IMPACT OF STUDY: NTM are emerging as important causative agents of pulmonary and extra pulmonary disease, the ability to recognize disease caused by NTM and subsequently treat such disease has become increasingly important. The identification of NTM up to its species level using HPLC and LPA CM/AS should gain importance in all TB reference Laboratories.

Recurrence of tuberculosis among newly diagnosed sputum positive pulmonary tuberculosis patients treated under the Revised National Tuberculosis Control Programme, India: A multi-centric prospective study.

INTRODUCTION: There is lack of information on the proportion of new smear-positive pulmonary tuberculosis (PTB) patients treated with a 6-month thrice-weekly regimen under Revised National Tuberculosis Control Programme (RNTCP) who develop recurrent TB after successful treatment outcome. OBJECTIVE: To estimate TB recurrence among newly diagnosed PTB patients who have successfully completed treatment and to document endogenous reactivation or re-infection. Risk factors for unfavourable outcomes to treatment and TB recurrence were determined. METHODOLOGY: Adult (aged ≥ 18 yrs) new smear positive PTB patients initiated on treatment under RNTCP were enrolled from sites in Tamil Nadu, Karnataka, Delhi, Maharashtra, Madhya Pradesh and Kerala. Those declared "treatment success" at the end of treatment were followed up with 2 sputum examinations each at 3, 6 and 12 months after treatment completion. MIRU-VNTR genotyping was done to identify endogenous re-activation or exogenous re-infection at TB recurrence. TB recurrence was expressed as rate per 100 person-years (with 95% confidence interval [95%CI]). Regression models were used to identify the risk factors for unfavourable response to treatment and TB recurrence. RESULTS: Of the1577 new smear positive PTB patients enrolled, 1565 were analysed. The overall cure rate was 77% (1207/1565) and treatment success was 77% (1210 /1565). The cure rate varied from 65% to 86%. There were 158 of 1210 patients who had TB recurrence after treatment success. The pooled TB recurrence estimate was 10.9% [95%CI: 0.2-21.6] and TB recurrence rate per 100 person-years was 12.7 [95% CI: 0.4-25]. TB recurrence per 100 person-years varied from 5.4 to 30.5. Endogenous reactivation was observed in 56 (93%) of 60 patients for whom genotyping was done. Male gender was associated with TB recurrence. CONCLUSION: A substantial proportion of new smear positive PTB patients successfully treated with 6 -month thrice-weekly regimen have TB recurrence under program settings.

Ligand-Dependent Modulation of G Protein Conformation Alters Drug Efficacy.

G protein-coupled receptor (GPCR) signaling, mediated by hetero-trimeric G proteins, can be differentially controlled by agonists. At a molecular level, this is thought to occur principally via stabilization of distinct receptor conformations by individual ligands. These distinct conformations control subsequent recruitment of transducer and effector proteins. Here, we report that ligand efficacy at the calcitonin GPCR (CTR) is also correlated with ligand-dependent alterations to G protein conformation. We observe ligand-dependent differences in the sensitivity of the G protein ternary complex to disruption by GTP, due to conformational differences in the receptor-bound G protein hetero-trimer. This results in divergent agonist-dependent receptor-residency times for the hetero-trimeric G protein and different accumulation rates for downstream second messengers. This study demonstrates that factors influencing efficacy extend beyond receptor conformation(s) and expands understanding of the molecular basis for how G proteins control/influence efficacy. This has important implications for the mechanisms that underlie ligand-mediated biased agonism. VIDEO ABSTRACT.

Perforated Gastric Gangrene without Pneumothorax in an Adult Bochdalek Hernia due to Volvulus.

Bochdalek hernia is the most frequent congenital diaphragmatic hernia which occurs due to a defect in the posterior attachment of the diaphragm when there is a failure of closure of the pleuroperitoneal membrane in utero. It rarely presents for the first time in adults. We report one such case of a 23-year-old male patient who presented with an acute abdomen. Chest X-ray showed air under diaphragm and he was taken up for an emergency laparotomy. Intraoperatively an organoaxial volvulus of the stomach was found in a bochdaleks hernia with a focal gangrene of the stomach fundus with perforation and peritonitis. However, there was no breach of pleural cavity. A sleeve resection of the gangrenous portion of the stomach was performed and the diaphragmatic defect was repaired. Patient made an uneventful postoperative recovery. Gastric gangrene with perforation as a manifestation of the adult bochdalek hernia is indeed rare. A concomitant pneumothorax occurs along with this condition which requires an intercostal drainage tube prior to the laparotomy. We report this case for its unique presentation without pneumothorax.

Cost analysis of different diagnostic algorithms for pulmonary tuberculosis varying in placement of Xpert MTB/RIF.

BACKGROUND: We undertook cost analysis for diagnosis of pulmonary tuberculosis (PTB) using present algorithm under Revised National Tuberculosis Control programme and using Xpert MTB/RIF (Xpert) as frontline test or in conjunction with smear microscopy and/or chest radiography. METHODS: Costs were estimated for different strategies: (A) present algorithm involving sputum smear examination followed by antibiotic trial in smear negative patients, repeat smear examination (RE) if symptoms continue and chest radiography if RE negative; (B) direct Xpert; (C) smear microscopy followed by Xpert in smear negative patients; (D) radiography followed by Xpert in those having abnormal pulmonary shadows; and (E) smear examination followed by radiography among smear negative patients and Xpert in presence of abnormal pulmonary shadow. RESULTS: Cost to program was estimated lowest with Strategy A and highest with Strategy B. Compared to the latter, program cost reduces by 7%, 4.5%, and 17.4% by strategies C, D, and E, respectively. Cost to the group of individuals with presumptive PTB and their attendants is significantly higher for Strategy A compared to other four strategies. Among the latter, the patients' cost was minimum with Strategy B and maximum with Strategy C. Program cost per case diagnosed was lowest by Strategy A and highest by Strategy B. Patient cost per case diagnosed was highest by Strategy A and lowest by Strategy B. Using Xpert, Strategy E had the lowest program as well as overall cost per case diagnosed. CONCLUSION: Strategy E may be chosen for diagnosis of PTB. When resources would no longer be a constraint, direct Xpert would reduce costs incurred by the patients.

A simple method to generate stable cell lines for the analysis of transient protein-protein interactions.

Transient protein-protein interactions form the basis of signal transduction pathways in addition to many other biological processes. One tool for studying these interactions is bioluminescence resonance energy transfer (BRET). This technique has been widely applied to study signaling pathways, in particular those initiated by G protein-coupled receptors (GPCRs). These assays are routinely performed using transient transfection, a technique that has limitations in terms of assay cost and variability, overexpression of interacting proteins, vector uptake limited to cycling cells, and non-homogenous expression across cells within the assay. To address these issues, we developed bicistronic vectors for use with Life Technology's Gateway and flpIN systems. These vectors provide a means to generate isogenic cell lines for comparison of interacting proteins. They have the advantage of stable, single copy, isogenic, homogeneous expression with low inter-assay variation. We demonstrate their utility by assessing ligand-induced interactions between GPCRs and arrestin proteins.

Patient-specific simulations of stenting procedures in coronary bifurcations: two clinical cases.

Computational simulations of stenting procedures in idealized geometries can only provide general guidelines and their use in the patient-specific planning of percutaneous treatments is inadequate. Conversely, image-based patient-specific tools that are able to realistically simulate different interventional options might facilitate clinical decision-making and provide useful insights on the treatment for each individual patient. The aim of this work is the implementation of a patient-specific model that uses image-based reconstructions of coronary bifurcations and is able to replicate real stenting procedures following clinical indications. Two clinical cases are investigated focusing the attention on the open problems of coronary bifurcations and their main treatment, the provisional side branch approach. Image-based reconstructions are created combining the information from conventional coronary angiography and computed tomography angiography while structural finite element models are implemented to replicate the real procedure performed in the patients. First, numerical results show the biomechanical influence of stents deployment in the coronary bifurcations during and after the procedures. In particular, the straightening of the arterial wall and the influence of two overlapping stents on stress fields are investigated here. Results show that a sensible decrease of the vessel tortuosity occurs after stent implantation and that overlapping devices result in an increased stress state of both the artery and the stents. Lastly, the comparison between numerical and image-based post-stenting configurations proved the reliability of such models while replicating stent deployment in coronary arteries.

Beyond mere markers: functions for CD34 family of sialomucins in hematopoiesis.

CD34, podocalyxin, and endoglycan are members of a family of single-pass transmembrane proteins that show distinct expression on early hematopoietic precursors and vascular-associated tissue. In spite of the fact that the expression of CD34 on these early progenitors has been known for over 20 yr and used clinically in hematopoietic stem cell transplantation for more than 15 yr, little is known about its exact role or function. More recently, CD34 expression has been shown to distinguish activated early progenitors from quiescent cells. With the subsequent identification of podocalyxin and endoglycan as related family members also expressed on early progenitor cells, attention is slowly shifting toward understanding how these molecules might contribute to progenitor function and behavior. In this review we examine the existing evidence and propose testable models to reveal the importance of these molecules for stem and progenitor cell function.