Mapping systemic lupus erythematosus heterogeneity at the single-cell level.

Patients with systemic lupus erythematosus (SLE) display a complex blood transcriptome whose cellular origin is poorly resolved. Using single-cell RNA sequencing, we profiled ~276,000 peripheral blood mononuclear cells from 33 children with SLE with different degrees of disease activity and 11 matched controls. Increased expression of interferon-stimulated genes (ISGs) distinguished cells from children with SLE from healthy control cells. The high ISG expression signature (ISGhi) derived from a small number of transcriptionally defined subpopulations within major cell types, including monocytes, CD4+ and CD8+ T cells, natural killer cells, conventional and plasmacytoid dendritic cells, B cells and especially plasma cells. Expansion of unique subpopulations enriched in ISGs and/or in monogenic lupus-associated genes classified patients with the highest disease activity. Profiling of ~82,000 single peripheral blood mononuclear cells from adults with SLE confirmed the expansion of similar subpopulations in patients with the highest disease activity. This study lays the groundwork for resolving the origin of the SLE transcriptional signatures and the disease heterogeneity towards precision medicine applications.

Circularly Polarized Luminescence and Circular Dichroism of Bichromophoric Difluoroboron-ß-diketonates: Inversion and Enhanced Chirality Based on Spatial Arrangements and Self-Assembly.

We synthesized two bichromophoric difluoroboron-ß-diketonates (DFB) connected in para and meta positions by using cyclohexane diamine as a chiral bridge (para and meta (R/S)-CyDFB). TD-DFT calculations revealed that the variation in connectivity of the DFB units leads to different spatial arrangements and a chirality inversion of the bichromophoric DFB. Higher gabs values were obtained in (R/S)-CyDFB connected in para as compared to meta position. Aggregation of para (R/S)-CyDFB in mixture of solvents increase the glum values as compared to its monomeric form. Ultrasonication and heating induced the formation of highly ordered nano-helical wires of para (R/S)-CyDFB that increased the glum values to 0.015. On the other hand, meta (R/S)-CyDFB failed to form highly ordered self-assembled wires due to hindered H-binding sites. These observations indicate that the chiroptical properties of DFB bi-chromophore system can be modulated with self-assembly and spatial arrangement of the chromophores.

Detection of Salmonella enterica subsp. enterica via Quenching of Unincorporated Amplification Signal Reporters in Loop-Mediated Isothermal Amplification.

Salmonella enterica is a major cause of diarrheal diseases in developing countries where timely surveillance and proper clinical management are inadequate. In this study, a rapid and cheap method of detecting S. enterica DNA was developed by employing the Quenching of Unincorporated Amplification Signal Reporters in Loop-Mediated Isothermal Amplification (QUASR LAMP) platform. QUASR LAMP provides a closed-tube, target-specific endpoint detection of pathogens, wherein results can be analyzed visually through an LED transilluminator and verified through agarose gel electrophoresis. Based on the chromosomal SopD gene, primers and probes were first designed, then screened. The assay was subsequently optimized so that the presence of S. enterica is determined by incubating the extracted DNA at 65°C for only 60 minutes. The assay was repeatable and can be performed by simply using a thermal cycler or even a dry bath incubator. S. enterica positives appear bright yellow green when viewed through a yellow filter excited with blue LED. The developed assay had an in silico and in vitro specificity towards Salmonella enterica subsp. enterica serovars with a limit of detection of 104 copies per microliter. The Salmonella QUASR LAMP assay has the potential for food and environmental applications. Chiefly, as an alternative to traditional microbiology and PCR, this QUASR LAMP assay can be used for point-of-care salmonellosis testing of clinical specimens in low-resource settings.

Behcet's-like syndrome following pembrolizumab: An immune-related adverse event associated with programmed death receptor-1 inhibitor therapy.

INTRODUCTION: The landscape for the treatment of metastatic melanoma has been revolutionized with the introduction immune checkpoint inhibitors. Immune checkpoint inhibitors have now become the standard of care for the treatment of cancers. These immune agents including programmed death receptor-1 inhibitors, programmed death-ligand 1 inhibitors and cytotoxic T-lymphocyte antigen-4 inhibitors have shown promising results but have been associated with numerous immune-related complications. Pembrolizumab, a programmed death receptor-1 inhibitor, has been associated with a number of immune-related adverse events affecting multiple organ systems including integument, ocular, endocrine, cardiovascular, pulmonary, renal, gastrointestinal, and musculoskeletal system. CASE REPORT: We present a case of an 88-year-old Caucasian male with metastatic melanoma of the face with metastasis to the right fifth cranial nerve and into the right cavernous sinus. He underwent resection of the melanoma and was placed on pembrolizumab at 2 mg/kg every three weeks. Interestingly, 24 months on pembrolizumab therapy, he developed corneal erosions, oral and genital ulcerations. MANAGEMENT AND OUTCOME: Patient completed his 24 months of pembrolizumab and was started on prednisone and colchicine with improvement in his symptoms. At his follow-up eight months, he had recurrence of an oral ulcer. DISCUSSION: Here we present a rare case of an elderly male on pembrolizumab who suffered from corneal erosions, oral and genital ulcers, a syndrome similar to Behcet's disease. Given that pembrolizumab and other immune checkpoint inhibitors are being utilized in the treatment of cancers, physicians should be aware of the wide range immune-related adverse events including the possible Behcet's-like syndrome presentation.

Characterizing fitness and immune escape of SARS-CoV-2 EG.5 sublineage using elderly serum and nasal organoid.

SARS-CoV-2 Omicron variant has evolved into sublineages. Here, we compared the neutralization susceptibility and viral fitness of EG.5.1 and XBB.1.9.1. Serum neutralization antibody titer against EG.5.1 was 1.71-fold lower than that for XBB.1.9.1. However, there was no significant difference in virus replication between EG.5.1 and XBB.1.9.1 in human nasal organoids and TMPRSS2/ACE2 over-expressing A549 cells. No significant difference was observed in competitive fitness and cytokine/chemokine response between EG.5.1 and XBB.1.9.1. Both EG.5.1 and XBB.1.9.1 replicated more robustly in the nasal organoid from a younger adult than that from an older adult. Our findings suggest that enhanced immune escape contributes to the dominance of EG.5.1 over earlier sublineages. The combination of population serum susceptibility testing and viral fitness evaluation with nasal organoids may hold promise in risk assessment of upcoming variants. Utilization of serum specimens and nasal organoid derived from older adults provides a targeted risk assessment for this vulnerable population.

Hematologic adverse events reported after COVID-19 vaccination in the Philippines: A national database study.

Vaccination is the most important strategy in preventing COVID-19. Vaccine efficacy and safety have been established in clinical trials but real-world data are useful to determine occurrence of adverse events in a population with heterogeneous characteristics. Knowledge on the hematologic events associated with different COVID-19 vaccines would be beneficial for patients as well as hematologists who oversee the care of these patients. This study aimed to determine the rates and outcomes of hematologic adverse events after COVID-19 vaccination in the Philippines. In this self-controlled case series, there were 268 individuals reported to have hematologic adverse events. Most received Comirnaty at 29.85%. Majority (62.31%) reported hematologic adverse events following the first dose of the vaccine. The overall event rate was 0.0182 per 10,000 vaccine doses; and lymphadenopathy was the most common hematologic adverse effect with a rate of 0.011 per 10,000 vaccine doses, followed by anemia at 0.0034 per 10,000 vaccine doses and thrombocytopenia at 0.0017 per 10,000 vaccine doses. Autoimmune cytopenias were also reported with an event rate of 0.0007 per 10,000 vaccine doses for ITP. One-hundred thirty two (49.25%) were fully recovered and 23.88% were recovering from hematologic adverse events as of the time of writing. The study showed a low rate of hematologic adverse events post COVID-19 vaccination with the seven different vaccine brands administered in the Philippines.

A six-month crossover chemoprevention clinical trial of tea in smokers and non-smokers: methodological issues in a feasibility study.

BACKGROUND: Chemoprevention crossover trials of tea can be more efficient than parallel designs but the attrition and compliance rates with such trials are unknown. METHODS: Attrition (dropouts) and compliance with treatment were assessed in a 25-week randomized, placebo controlled, crossover, feasibility clinical trial of four tea treatments to investigate the effect of tea on oral cancer biomarkers. Each treatment lasted 4 weeks with 2 weeks of washout in between. Participants were 32 smokers and 33 non-smokers without any evidence of premalignant oral lesions. The interventions consisted of packets of green tea, black tea, caffeinated water, or placebo. Participants were assigned to each treatment for four weeks, and were instructed to drink five packets per day while on the treatment. Dropout from the trial and compliance (consumption of ≥85% of the prescribed treatment packets) are the main outcome measures reported. RESULTS: There was a high rate of dropout (51%) from the study, and the rates were significantly higher among smokers (64%) than non-smokers (36%). Among participants who completed the study the rate of compliance was 72%. The highest rates of dropouts occurred between the first and second treatment visits in both smokers (38% dropout) and non-smokers (18% dropout). Throughout the study smokers were more likely to dropout than non-smokers. Black tea treatment was associated with the highest rates of dropout among smokers (37%), but was associated with the lowest rate of dropout among non-smokers (4%). CONCLUSIONS: In a study conducted to test the feasibility of a four-treatment crossover tea trial, a high rate of dropout among smokers and non-smokers was observed. Multi-arm crossover tea trials might pose a higher burden on participants and research is needed to improve adherence and treatment compliance in such trials. TRIAL REGISTRATION NUMBER: ISRCTN70410203.

Evaluation of loop-mediated isothermal amplification assay and enzyme-linked immunosorbent assay in detecting Schistosoma japonicum in Siargao Island, Surigao del Norte, the Philippines.

OBJECTIVES: This study aimed to describe the prevalence and seroprevalence of schistosomiasis in Siargao Island, Surigao del Norte and to compare the performance of enzyme-linked immunosorbent assay antibody test (ELISA Ab) and loop-mediated isothermal amplification assay (LAMP) for diagnosis of schistosomiasis. METHODS: The study was conducted in selected barangays (villages) in five municipalities in Siargao Island, Surigao del Norte and included school-age children (SAC) who submitted stool and blood samples. Stool samples were examined using the Kato-Katz technique. Blood samples were collected through venipuncture. The stool samples and the blood samples collected were tested using LAMP assay and polymerase chain reaction (PCR). The blood samples were examined using ELISA Ab. Diagnostic performance of LAMP assay using stool specimen was evaluated using Kato-Katz technique and PCR assay as the composite reference standard, while PCR assay was used as the reference standard to evaluate LAMP assay and ELISA Ab using blood specimens. RESULTS: A total of 417 stool samples from SAC were examined. The prevalence of schistosomiasis and moderate-heavy intensity (MHI) schistosomiasis were 3.8% and 1.4%, respectively. Schistosomiasis and soil-transmitted helminthiases (STH) coinfection prevalence were 2.6%. A total of 425 blood samples were examined using ELISA Ab. Seroprevalence was 61.6%. The municipality of San Isidro had the highest seroprevalence at 84.8%, while Burgos had the lowest seroprevalence at 48.5%.LAMP assay had higher sensitivity and positive predictive value but lower specificity when using stool than when using blood samples. Its negative predictive value was similar regardless of the specimen used. ELISA Ab has higher sensitivity and negative predictive value than LAMP assay although it has lower specificity and positive predictive value. This may be due to ELISA Ab measuring Schistosoma exposure and is thus unable to distinguish past from active infection. CONCLUSIONS: Schistosomiasis remains a public health concern in Siargao Island, Surigao del Norte. The locally developed LAMP assay offers a simpler diagnostic test for schistosomiasis compared with PCR, while minimizing the risk of misdiagnosis compared with Kato-Katz technique. It could serve as a point of care diagnostics for schistosomiasis. ELISA Ab is more useful in surveillance particularly in low-endemicity areas where determination of exposure is more important than differentiating past from active infection. ELISA Ab may be helpful in the clinical setting when coupled with the expertise of a physician who is familiar with schistosomiasis.

A CTGF-YAP Regulatory Pathway Is Essential for Angiogenesis and Barriergenesis in the Retina.

Connective tissue growth factor (CTGF) or cellular communication network 2 (CCN2) is a matricellular protein essential for normal embryonic development and tissue repair. CTGF exhibits cell- and context-dependent activities, but CTGF function in vascular development and barrier function is unknown. We show that endothelial cells (ECs) are one of the major cellular sources of CTGF in the developing and adult retinal vasculature. Mice lacking CTGF expression either globally or specifically in ECs exhibit impaired vascular cell growth and morphogenesis and blood barrier breakdown. The global molecular signature of CTGF includes cytoskeletal and extracellular matrix protein, growth factor, and transcriptional co-regulator genes such as yes-associated protein (YAP). YAP, itself a transcriptional activator of CTGF, mediates several CTGF-controlled angiogenic and barriergenic transcriptional programs. Re-expression of YAP rescues, at least partially, angiogenesis and barriergenesis in CTGF mutant mouse retinas. Thus, the CTGF-YAP regulatory loop is integral to retinal vascular development and barrier function.

Digital malaria information sharing in Asia.

In 2001 a project named Mekong Malaria Documentation Center (MMDC) was established in Vietnam to support the information needs of malaria professionals in the Mekong Region. In 2005, MMDC was renamed and reorganized into a new database called ACTMalaria Information Resource Center (AIRC). AIRC's goal is to gather all published and unpublished materials, including electronic files, regarding malaria from ACTMalaria member countries. Today, there are eleven member nations. More than 1,500 files have been uploaded and are available for access. Various areas of AIRC still need to be developed, including editing, approvals of files, coordination with publishers, copyright issues, and the architecture of the database. Funding also needs to be secured for the sustainability of AIRC.

Psychiatrists' perceptions and practices in treating patients' obesity.

OBJECTIVE: This study identified psychiatrists' perceptions and practices regarding advising and treating obese patients. METHODS: Questionnaires were mailed to a national random sample of 500 members of APA. A three-wave mailing was used to maximize the return rate. The questionnaire contained items on weight control based on the Stages of Change and Health Belief models, Self-Efficacy theory, and the 5As strategy. RESULTS: A total of 236 psychiatrists responded to the survey. Most did not have any formal training during medical school on treating obese patients and three-quarters evaluated their training in psychiatric residency programs on weight loss/control issues as "not adequate at all" or "not very adequate." The majority regularly assisted their obese patients with weight management. Most psychiatrists felt confident to ask, advise, assess, assist, and arrange (5As) regarding weight loss issues and believed that doing so would result in significant weight loss. The majority were more likely to advise obese patients to lose weight when comorbid conditions were present. The most common barriers to aiding obese patients were time constraints, poor patient compliance, lack of clear guidelines and practice tools, limited medical training on the issue, and fear of offending patients. CONCLUSION: Patients can no longer afford to have their psychiatrists provide cursory assistance with obesity. Weight management training should be incorporated into psychiatric residency training and continuing education programs.

Adalimumab-induced Anti-neutrophilic Cytoplasmic Antibody Vasculitis: A Rare Complication of an Increasingly Common Treatment.

Tumor necrosis factor (TNF) inhibitors are used for treatment of different autoimmune diseases. Interestingly they are also being noted to cause autoimmune side effects such as vasculitis, systemic lupus erythematosus, interstitial lung disease, etc. We describe one such case of granuloma annulare being treated with Adalimumab, who then developed pulmonary-renal syndrome form anti-neutrophilic cytoplasmic antibody (ANCA)-induced vasculitis. This was treated with steroids and immunosuppression, as well as discontinuation of the TNF inhibitor. However, he remains dependant on dialysis and immunosuppression. In this article, we review the existing literature on clinical presentation and course of TNF inhibitors-induced ANCA vasculitis. We also discuss the underlying mechanisms thought to be responsible for this.

Genotypic persistence of dengue virus in the Philippines.

The Philippines is known to have one of the world's highest prevalences of dengue infection. The disease has been endemic in the country since 1956 and the severe form was first reported during an outbreak in Manila in 1954. Among all of the countries in the world, the Philippines had the highest case fatality rate from 2008 to 2012. With the increasing rate of international travel, the country is also considered one of the primary sources of imported dengue cases in non-endemic areas in Asia, Australia, and Europe. Despite this high prevalence, there is a dearth of literature describing the circulating strains in the Philippines at the genotype level. Using data from sequence databases, this study aimed to characterize all available Philippine sequences, at the molecular level. Capsid/pre-membrane (C/prM) junction gene and envelope (E) gene sequences of dengue serotypes 1, 2, 3 and 4 from 1956 to 2016 were used for phylogenetic analysis and genotypic identification. All four serotypes co-circulate in the country over the last 50 years with conspicuous genotypic characteristics. DENV-1 exhibited an apparent persistence of a single genotype since 1974. DENV-2 showed strong evidence of genotypic shift in 1999-2002 accompanied by a genotypic persistence thereafter. DENV-3 and DENV-4 displayed a temporal domination of a single genotype, with evidence of a minor co-circulating genotypic population. The persistence and pre-domination of specific DENV genotypes warrant continuous molecular surveillance for signs of genotypic shifts that can cause local outbreak events or an increased risk for severity.

"It's like being in another world": demonstrating the benefits of therapeutic horseback riding for individuals with psychiatric disability.

Five adults with longstanding histories of psychiatric disabilities were recruited for a ten-week therapeutic horseback riding program. Individuals learned basic riding skills and had the opportunity to bond with a horse. In addition, the riders participated in a post-riding process group that used artistic and creative exercises to promote individual expression. By the end often weeks, the riders reported success in learning basic horsemanship and, in doing so, also reported additional psychosocial benefits, including an augmented sense of self-efficacy and self-esteem. In sum, this adjunctive therapy can facilitate the recovery process.

Continued circulation of a single genotype of dengue virus serotype 2 in the Philippines.

OBJECTIVE: To obtain descriptive information of behavioral pattern in Chinese school-aged children with cleft lip and palate. METHODS: A total of 93 cleft lip and palate patients between the age of 6-11 year-old and treated at West China Stomatology Hospital were selected. And another 100 unaffected controls, matched for age and gender, were recruited randomly from a common primary school in Chengdu. Chart review of medical records was used to obtain psychosocial checklists. Scores were compared with published norms and controls to evaluate the risk of problems, separately for three diagnostic groups. RESULTS: The patients group had lower scores of social and academic competencies, especially those with facial deformity or speech problem. No difference was found in the aspect of activity competency. All patients showed elevations in behavior problems. But the type of behavior problems varied in different genders. CONCLUSIONS: Chinese school-aged children with cleft lip and palate are at raised risk for social and academic difficulties. Specific pattern of behavior problems displays differently depending on gender of the patient.

Preliminary evaluation of in vitro cytotoxicity and in vivo antitumor activity of Xanthium strumarium in transplantable tumors in mice.

In the present study, active fractions of the methanolic extract of Xanthium strumarium (XS) showing potent cytotoxicity were determined using microculture tetrazolium (MTT) and sulforhodamine B (SRB) assays in selected cancer cell lines. The active fractions viz., chloroform soluble fraction of root (CEXSR), hexane soluble fraction of leaf (HEXSL), hexane soluble fraction of fruits (HEXSF) and chloroform soluble fraction of fruits (CEXSF) of XS were tested in transplantable animal tumor models for their antitumor potential. Dalton's ascitic lymphoma (DLA) cells were used to induce solid and liquid (ascites) tumor in mice. The tumor bearing animals were treated with active fractions at two dose levels (100 and 200 mg/kg). The antitumor activities of the active fractions in tumor bearing animals were monitored with parameters such as body weight and increase in life-span as well as biochemical and hematological modalities (in the case of liquid tumor). Tumor incidence and tumor volume were the parameters monitored in the case of the solid tumor model. The results were analyzed by one-way ANOVA followed by Tukey's post hoc test. The extracts were found to increase the life-span of tumor bearing animals and restore the altered hematological and biochemical parameters significantly.

Genomic anatomy of the Tyrp1 (brown) deletion complex.

Chromosome deletions in the mouse have proven invaluable in the dissection of gene function. The brown deletion complex comprises >28 independent genome rearrangements, which have been used to identify several functional loci on chromosome 4 required for normal embryonic and postnatal development. We have constructed a 172-bacterial artificial chromosome contig that spans this 22-megabase (Mb) interval and have produced a contiguous, finished, and manually annotated sequence from these clones. The deletion complex is strikingly gene-poor, containing only 52 protein-coding genes (of which only 39 are supported by human homologues) and has several further notable genomic features, including several segments of >1 Mb, apparently devoid of a coding sequence. We have used sequence polymorphisms to finely map the deletion breakpoints and identify strong candidate genes for the known phenotypes that map to this region, including three lethal loci (l4Rn1, l4Rn2, and l4Rn3) and the fitness mutant brown-associated fitness (baf). We have also characterized misexpression of the basonuclin homologue, Bnc2, associated with the inversion-mediated coat color mutant white-based brown (B(w)). This study provides a molecular insight into the basis of several characterized mouse mutants, which will allow further dissection of this region by targeted or chemical mutagenesis.