Cryptococcal Pneumonia in a Patient on Tyrosine Kinase Inhibitor Therapy: How Common Is It?

Cryptococcal pneumonia is identified as a fungal infection of the lungs, with Cryptococcus neoformans and Cryptococcus gattii as the most common culprits. Cryptococcus neoformans primarily affects immunocompromised individuals while Cryptococcus gattii infections occur mostly in immunocompetent hosts. We present a 76-year-old male on ibrutinib due to a history of chronic lymphocytic leukemia who had multiple hospitalizations for pneumonia and was later diagnosed with cryptococcal pneumonia through positive bronchoalveolar lavage fungal culture and lymph node biopsy.

Protumorigenic role of the atypical cadherin FAT1 by the suppression of PDCD10 via RelA/miR221-3p/222-3p axis in glioblastoma.

The atypical cadherin FAT1 function either as a pro or antitumorigenic in tumors of different tissue origins. Our group previously demonstrated the protumorigenic nature of FAT1 signaling in glioblastoma (GBM). In this study, we investigated how FAT1 influences the expression of clustered oncomiRs (miR-221-3p/miR-222-3p) and their downstream effects in GBM. Through several experiments involving the measurement of specific gene/microRNA expression, gene knockdowns, protein and cellular assays, we have demonstrated a novel oncogenic signaling pathway mediated by FAT1 in glioma. These results have been verified using antimiRs and miR-mimic assays. Initially, in glioma-derived cell lines (U87MG and LN229), we observed FAT1 as a novel up-regulator of the transcription factor NFκB-RelA. RelA then promotes the expression of the clustered-oncomiRs, miR-221-3p/miR-222-3p, which in turn suppresses the expression of the tumor suppressor gene (TSG), PDCD10 (Programmed cell death protein10). The suppression of PDCD10, and other known TSG targets (PTEN/PUMA), by miR-221-3p/miR-222-3p, leads to increased clonogenicity, migration, and invasion of glioma cells. Consistent with our in-vitro findings, we observed a positive expression correlation of FAT1 and miR-221-3p, and an inverse correlation of FAT1 and the miR-targets (PDCD10/PTEN/PUMA), in GBM tissue-samples. These findings were also supported by publicly available GBM databases (The Cancer Genome Atlas [TCGA] and The Repository of Molecular Brain Neoplasia Data [Rembrandt]). Patients with tumors displaying high levels of FAT1 and miR-221-3p expression (50% and 65% respectively) experienced shorter overall survival. Similar results were observed in the TCGA-GBM database. Thus, our findings show a novel FAT1/RelA/miR-221/miR-222 oncogenic-effector pathway that downregulates the TSG, PDCD10, in GBM, which could be targeted therapeutically in a specific manner.

Deep Learning Classification of Tuberculosis Chest X-rays.

Background Tuberculosis (TB) is an infectious disease caused by the bacterium Mycobacterium tuberculosis. It primarily affects the lungs but can also affect other organs, such as the kidneys, bones, and brain. TB is transmitted through the air when an infected individual coughs, sneezes, or speaks, releasing tiny droplets containing the bacteria. Despite significant efforts to combat TB, challenges such as drug resistance, co-infection with human immunodeficiency virus (HIV), and limited resources in high-burden settings continue to pose obstacles to its eradication. TB remains a significant global health challenge, necessitating accurate and timely detection for effective management.  Methods This study aimed to develop a TB detection model using chest X-ray images obtained from Kaggle.com, utilizing Google's Collaboration Platform. Over 1196 chest X-ray images, comprising both TB-positive and normal cases, were employed for model development. The model was trained to recognize patterns within the TB chest X-rays to efficiently recognize TB within patients in order to be treated in time. Results The model achieved an average precision of 0.934, with precision and recall values of 94.1% each, indicating its high accuracy in classifying TB-positive and normal cases. Sensitivity and specificity values were calculated as 96.85% and 91.49%, respectively. The F1 score was also calculated to be 0.941. The overall accuracy of the model was found to be 94%.  Conclusion These results highlight the potential of machine learning algorithms for TB detection using chest X-ray images. Further validation studies and research efforts are needed to assess the model's generalizability and integration into clinical practice, ultimately facilitating early detection and improved management of TB.

Pneumoperitoneum in a Patient With COVID-19 and Acute Respiratory Distress Syndrome Without Pneumothorax.

We present a case report of pneumoperitoneum, pneumomediastinum, and subcutaneous emphysema in a patient with COVID-19 pneumonia-causing acute respiratory distress syndrome (ARDS) without any pneumothorax occurring. Pneumothorax, pneumomediastinum, and subcutaneous emphysema are known complications of barotrauma due to positive pressure from mechanical ventilation which is necessary for patients suffering from a severe case of COVID-19. In our literature search, we could not find any reported case of pneumoperitoneum without pneumothorax occurring. Our case is an important addition to the literature presenting a rare complication of mechanical ventilation in patients with ARDS.

Incidence of Barotrauma in COVID-19 Patients Requiring Mechanical Ventilation: A Retrospective Study in a Community Hospital.

Background and aim Acute respiratory distress syndrome (ARDS) is a severe complication of COVID-19 and traditional ventilation strategies using ARDSNet protocol, including low tidal volumes, appear to cause barotrauma in COVID-19 patients at a higher rate than non-COVID-19 ARDS patients. The purpose of our study was to determine if COVID-19 patients with ARDS undergoing mechanical ventilation at St. Joseph's Medical Center (SJMC) developed barotrauma at a higher rate than non-COVID-19 ARDS patients. Methods and materials This study was a retrospective chart review of all patients admitted to critical care units at SJMC with COVID-19 infection and requiring mechanical ventilation from March 1, 2020 to September 30, 2020. The sample included adult patients (aged 18 and above) with the International Classification of Diseases (ICD) 10 code for COVID-19 (U07.1) and patients who were placed on mechanical ventilation for longer than 24 hours, from March 1, 2020 to September 30, 2020. Barotrauma was confirmed via radiographic imaging including chest X-ray, CT, or CT angiography (CTA).  Results One hundred and forty COVID-19 patients underwent mechanical ventilation for longer than 24 hours from March 1, 2020 to September 30, 2020 at our facility. Twenty-six COVID-19 patients (18.6%) met our inclusion criteria (development of barotrauma during hospital admission) of which 25 patients (17.9%) underwent mechanical (invasive and/or non-invasive) ventilation prior to the development of barotrauma. Around 80% of the patients were on non-invasive mechanical ventilation prior to intubation and invasive mechanical ventilation. The categorical breakdown of barotrauma was as follows: pneumothorax 65.4%, subcutaneous emphysema 61.5%, pneumomediastinum 34.6%, and pneumoperitoneum 7.7%. None of the included patients had any previous history of documented barotrauma. Prior to the time of barotrauma, 17 patients were on volume control, seven were on pressure control, and one was not on mechanical ventilation. Of the 17 patients on volume control, only one patient was above the ARDSNet guideline of 6-8 mL/kg ideal body weight (IBW). In comparison to ARDS patients at SJMC in 2019, only two out of 28 patients (7.14%) developed barotrauma during mechanical ventilation.  Conclusions Patients with COVID-19 who underwent mechanical ventilation developed barotrauma at a higher rate than traditional non-COVID-19 patients with ARDS.

Upregulation of Atypical Cadherin FAT1 Promotes an Immunosuppressive Tumor Microenvironment via TGF-ß.

FAT atypical cadherin 1 (FAT1) promotes glioblastoma (GBM) by promoting protumorigenic inflammatory cytokine expression in tumor cells. However, tumors also have an immunosuppressive microenvironment maintained by mediators such as transforming growth factor (TGF)-ß cytokines. Here, we have studied the role of FAT1 in tumor immune suppression. Our preliminary TIMER2.0 analysis of The Cancer Genome Atlas (TCGA) database revealed an inverse correlation of FAT1 expression with infiltration of tumor-inhibiting immune cells (such as monocytes and T cells) and a positive correlation with tumor-promoting immune cells [such as myeloid-derived suppressor cells (MDSCs)] in various cancers. We have analyzed the role of FAT1 in modulating the expression of TGF-ß1/2 in resected human gliomas, primary glioma cultures, and other cancer cell lines (U87MG, HepG2, Panc-1, and HeLa). Positive correlations of gene expression of FAT1 and TGF-ß1/2 were observed in various cancers in TCGA, Glioma Longitudinal Analysis Consortium (GLASS), and Chinese Glioma Genome Atlas (CGGA) databases. Positive expression correlations of FAT1 were also found with TGF-ß1/2 and Serpine1 (downstream target) in fresh-frozen GBM samples using q-PCR. siRNA-mediated FAT1 knockdown in cancer cell lines and in primary cultures led to decreased TGF-ß1/2 expression/secretion as assessed by q-PCR, Western blotting, and ELISA. There was increased chemotaxis (transmigration) of THP-1 monocytes toward siFAT1-transfected tumor cell supernatant as a consequence of decreased TGF-ß1/2 secretion. Reduced TGF-ß1 expression was also observed in THP-1 cultured in conditioned media from FAT1-depleted glioma cells, thus contributing to immune suppression. In U87MG cells, decreased TGF-ß1 upon FAT1 knockdown was mediated by miR-663a, a known modulator. FAT1 expression was also observed to correlate positively with the expression of surrogate markers of MDSCs [programmed death ligand-1 (PD-L1), PD-L2, and interleukin (IL)-10] in glioma tumors, suggesting a potential role of FAT1 in MDSC-mediated immunosuppression. Hence, our findings elaborate contributions of FAT1 to immune evasion, where FAT1 enables an immunosuppressive microenvironment in GBM and other cancers via TGF-ß1/2.

Impact of C1q fixing donor-specific antibodies on renal transplant outcome.

Not all anti-HLA donor-specific antibodies (HLA-DSAs) are detrimental to renal allograft. In this context, the C1q complement activating ability of antibodies appears to be an important parameter to distinguish clinically inert versus detrimental DSAs. We evaluated sera of 206 consecutive primary live donor renal transplant recipients before transplant and at post-operative day 7, 30, 90, 180 and at the time of graft dysfunction for quantifying HLA-DSAs using single antigen bead assay on a Luminex platform. Patients positive for these antibodies with an MFI >500 were further screened for C1q fixing nature of DSA. Fourteen of the 18 antibody-positive patients had C1q fixing DSA with MFI value >5000. Only 4 antibody-positive patients did not have C1q fixing DSA. The MFI values of DSA detected by C1q assay were generally higher at least by 25% than those detected by the conventional IgG-SAB assay. Twelve of the 14 patients (85.71%) with C1q+ DSA developed antibody-mediated rejection during the mean follow-up period of 21.43 ± 8.03 months as compared to none of the four C1q-negative DSA (85.71% vs 0%; P = .001). These results suggest deleterious effect of C1q+ DSA vis-à-vis C1q-negative DSA on renal allograft.

Randomised controlled trials for COVID-19: evaluation of optimal randomisation methodologies-need for data validation of the completed trials and to improve ongoing and future randomised trial designs.

During the emerging COVID-19 (coronavirus disease 2019) pandemic, initially there were no proven treatment options. With the release of randomised controlled trial (RCT) results, we are beginning to see possible treatment options for COVID-19. The RECOVERY trial showed an absolute risk reduction in mortality by 2.8% with dexamethasone, and the ACTT-1 trial showed that treatment with remdesivir reduced the time to recovery by 4 days. Treatment with hydroxychloroquine (HCQ) and lopinavir/ritonavir did not show any mortality benefit in either the RECOVERY or World Health Organization (WHO) Solidarity trials. The National Institutes of Health (NIH) and Brazilian HCQ trials did not show any benefit for HCQ based on the seven-point ordinal scale outcomes. The randomisation methodologies utilised in these controlled trials and the quality of published data were reviewed to examine their adaptability to treat patients. We found that the randomisation methodologies of these trials were suboptimal for matching the studied groups based on disease severity among critically-ill hospitalised COVID-19 patients with high mortality rates. The published literature is very limited regarding the disease severity metrics among the compared groups and failed to show that the data are without fatal sampling errors and sampling biases. We also found that there is a definite need for the validation of data in these trials along with additional important disease severity metrics to ensure that the trials' conclusions are accurate. We also propose proper randomisation methodologies for the design of RCTs for COVID-19 as well as guidance for the publication of COVID-19 trial results.

Clinical relevance of major histocompatibility complex class I chain-related molecule A (MICA) antibodies in live donor renal transplantation - Indian Experience.

Antibody-mediated rejections (AMR) in the absence of circulating anti-HLA-DSA have highlighted the role of non-HLA antibodies, particularly those directed against endothelial cells. Of these, MICA (major histocompatibility complex class I chain-related molecule A) antibodies are the most notable and important because of their potential in promoting graft rejections. Limited studies have focused on the impact of MICA donor-specific antibodies (DSA) on graft outcome as compared to those that are not donor-specific (NDSA). We evaluated pre- and post-transplant sera at POD 7, 30, 90, 180 and the time of biopsy from 206 consecutive primary live donor renal transplant recipients for anti-MICA and anti-HLA antibodies using single antigen bead assay on a Luminex platform. Recipients who developed MICA antibodies and their donors were phenotyped for MICA alleles. For the purpose of antibody analysis, patients were categorized into three major groups: biopsy-proven AMR, acute cellular rejection (ACR) and those with no rejection episodes (NRE). During the mean follow-up period of 17.37 ± 6.88 months, 16 of the 206 recipients developed AMR, while ACR was observed in only 13 cases. A quarter (25%) of the AMR cases had anti-MICA antibodies as compared to 7.7% of those experiencing ACR and 6.2% of the NRE group. Allelic typing revealed that all MICA Ab +ve AMR cases were due to the presence of donor-specific antibodies. MICA-DSA even in the absence of HLA-DSA was significantly associated with AMR but not with ACR when compared with the NRE group (P = <.01).

Soluble Major Histocompatibility Complex Class I related Chain A (sMICA) levels influence graft outcome following Renal Transplantation.

BACKGROUND: Since soluble isoforms of MICA play an important role in modulating the immune response, we evaluated a possible correlation between their levels and development of acute rejection following renal transplantation. METHODS: Serum samples collected at pre- and different time points post-transplant from 137 live related donor renal transplant recipients were evaluated retrospectively for sMICA levels and for the presence of MICA antibodies. Samples from 30 healthy volunteers were also tested as controls. RESULTS: Significantly higher levels of sMICA were observed in the pretransplant sera of allograft recipients as compared to healthy controls. Patients with acute cellular rejection experienced a significant fall in their levels at the time of diagnosis as compared to their pretransplant values and posttransplant follow up time points (p = .01, .003, .005 and .04 respectively at pre vs biopsy (Bx), POD7 vs Bx, POD 30 vs Bx, POD 90 vs Bx). However, no such difference was noted in patients undergoing antibody mediated rejection. Further the study did not reveal any correlation on the presence/absence of MICA antibodies with either an increase or decrease in sMICA levels. CONCLUSIONS: Estimating circulating levels of soluble MICA could provide useful information of prognostic importance in assessing graft outcome following renal transplantation.

Comparative analysis of Luminex-based donor-specific antibody mean fluorescence intensity values with complement-dependent cytotoxicity & flow crossmatch results in live donor renal transplantation.

BACKGROUND & OBJECTIVES: Antibodies specific to donor human leucocyte antigen (HLA) play a critical role in graft rejection and graft loss. In recent years, techniques for their detection have evolved significantly providing an ever-increasing degree of sensitivity and specificity, from the conventional cell-based assays to the advanced solid-phase system based on the Luminex platform. Consensus is still evolving on the routine employment of all these methods, either stand alone or in combination. The objective of this study was to explore the near-accurate mean fluorescence intensity (MFI) cut-off values detected on Luminex platform predicting the strength of cell-based crossmatch results. METHODS: Serum samples from 116 primary renal transplant recipients awaiting transplantation were tested for the presence of antidonor antibodies by the complement-dependent cytotoxicity (CDC) and flow crossmatch (FCXM) methods with their corresponding donors as well as for HLA-donor-specific antibodies (DSA) detection using a sensitive single antigen bead (SAB) assay. RESULTS: None of the patients having HLA Class I DSA with MFI values <1000 showed positivity for T-cell FCXM or CDC crossmatch, while in the group having MFI values between 1000 and 3000, 54 per cent showed positivity for the FCXM but none by the CDC method. However, in the group having MFI values >3000, 95 per cent of cases were positive for FCXM. Further, those groups with MFI values between 3000 and 5000, only 36 per cent were positive for CDC crossmatch, while 90 per cent showed positivity in the group with MFI >7000. INTERPRETATION & CONCLUSIONS: A cut-off MFI value of 3000 for Luminex SAB-based assay was found to significantly correlate with the FCXM positivity while a MFI value of 7000 and above predicted a positive CDC crossmatch. MFI cut-off value obtained as a surrogate marker for CDC and FCXM tests will help in resolving the limitations of different cell-based techniques.

Clinical relevance of antibody development in renal transplantation.

The detection and characterization of anti-HLA antibodies and the clinical impact of their appearance following renal transplantation are areas of immense interest. In particular, de novo development of donor-specific antibodies (DSA) has been associated with acute and chronic antibody-mediated graft rejection (AMR). Recently, methods for antibody detection have evolved remarkably from conventional cell-based assays to advanced solid phase systems. These systems have revolutionized the art of defining clinically relevant antibodies that are directed toward a renal graft. While anti-HLA DSAs have been widely associated with poor graft survival, the role of non-HLA antibodies, particularly those directed against endothelial cells, is beginning to be realized. Appreciation of the mechanisms underlying T cell recognition of alloantigens has generated great interest in the use of synthetic peptides to prevent graft rejection. Hopefully, continued progress in unraveling the molecular mechanisms of graft rejection and posttransplant monitoring of antibodies using highly sensitive testing systems will prove beneficial to immunological risk assessment and early prediction of renal allograft failure.

Management of calcium channel antagonist overdose with hyperinsulinemia-euglycemia therapy: case series and review of the literature.

Calcium channel antagonists (CCAs) are commonly involved in drug overdoses. Standard approaches to the management of CCA overdoses, including fluid resuscitation, gut decontamination, administration of calcium, glucagon, and atropine, as well as supportive care, are often ineffective. We report on two patients who improved after addition of hyperinsulinemia-euglycemia (HIE) therapy. We conclude with a literature review on hyperinsulinemia-euglycemia therapy with an exploration of the physiology behind its potential use.

How far have we reached in tuberculosis vaccine development?

Tuberculosis, a bacterial disease prevalent since ancient times, continues to cause the most deaths globally compared with all other diseases. The causative agent Mycobacterium tuberculosis is responsible for different types of tuberculosis in humans; however, pulmonary tuberculosis is the most common and causes the most deaths. Mycobacterium tuberculosis is an intracellular pathogenic bacterium, which has developed sophisticated mechanisms to survive inside host mononuclear phagocytes and thus evade the host immune system. This is attributed primarily to an inadequate immune response toward infecting bacteria, which results in temporary growth inhibition rather than death and subsequently allows the bacteria to multiply immensely, leading to full-blown disease in an individual. This disease has become a challenge due to poor diagnosis, a low-efficiency tuberculosis vaccine (Mycobacterium bovis Bacillus Calmette-Guerin [BCG]), a long-term antibacterial chemotherapy regimen (approximately 6 months), and an emergence of multiple drug resistant strains of Mycobacterium tuberculosis especially in people with human immune deficiency virus (HIV) infection, for whom researchers worldwide must develop effective short-term chemotherapy and an effective vaccine. In this review different aspects of vaccines in tuberculosis are discussed, and these include the traditional BCG vaccine, the modern auxotrophic vaccine, the subunit or acellular vaccine; and a DNA vaccine. We discuss also the potential of mycobacterial lipids as a vaccine or as an adjuvant in the future. Since complete genome information of Mycobacterium tuberculosis H37Rv and bioinformatics tools are available, it is possible to develop new strategies for a better and effective tuberculosis vaccine, which can replace the traditional BCG vaccine.