Faculty of Medical Siriraj Hospital

Noninvasive molecular tests of urine cells have been developed to monitor the activity of kidney diseases. We evaluate whether measurement of urinary messenger RNA (mRNA) levels of chemokine and growth factor genes could distinguish between diffuse proliferative lupus nephritis (class IV LN) and others and whether it is able to predict the response to therapy. Prebiopsy urine samples were collected

Human skin keratinocytes HaCat attacked by Staphylococcus aureus α-toxin showed a transient drop of cellular ATP levels whereas in toxin-perforated bovine mammary epithelial cells (BMEC), the ATP levels dropped more slowly. Morphologically, during the ATP level depletion, HaCat cell developed a spacious intracellular vacuole together with the transient influx of trypan blue. WST-1 signal, which tested the function of mitochondrial enzyme in viable cells, also decreased concomitantly. On the other hand, BMEC excluded trypan blue and vacuolation was not observed throughout the experiment. We conclude that mammary epithelial cells resist the toxin better than keratinocytes. This is the first report showing that α-toxin enhances transient membrane permeability to large molecules, temporary vacuole formation and the transient defect of mitochondrial enzyme in viable cells without cell lysis.Les cellules HaCat dérivées de kératinocytes humains infectées par Staphylococcus aureus α-toxin ont montré une chute temporaire du taux cellulaire d’ATP, alors que dans les cellules épithéliales mammaires bovines (CEMB) infectées, le taux cellulaire d’ATP a chuté plus lentement. Morphologiquement, durant la chute du taux d’ATP, les cellules HaCat ont développé une spacieuse vacuole intracellulaire ainsi qu’un afflux temporaire du bleu trypan. Le signal WST-1, qui évaluait l’activité de l’enzyme mitochondriale dans les cellules viables, a aussi dimunué simultanément. En revanche, au fil de l’expérience, aucune exclusion du bleu trypan ni une formation de vacuole n’ont été remarquées dans les CEMB. Nous concluons que les cellules épithéliales mammaires résistent mieux à la toxine que les kératinocytes. Cette étude est le premier rapport qui montre que l’α-toxin permet la perméabilité membranaire temporaire aux grandes molécules, une formation temporaire de vacuole et un défaut passager de l’enzyme mitochondriale dans les cellules viables sans lyse cellulaire.

Objective-Functionally interactive proteases of the plasminogen/plasmin and the matrix metalloproteinase (MMP) system degrade and reorganize the extracellular matrix of the vessel wall in atherosclerosis. Here we investigated whether such proteases are able to confer atherogenic properties onto low density lipoprotein by nonoxidative modification. Methods and Results-Similar to the recently described enzymatically-modified low-density lipoprotein (E-LDL), native LDL exposed to plasmin or matrix MMP-2 or MMP-9 and cholesterylester-hydrolase (CEH) showed extensive deesterification, with ratios of free cholesterol to total cholesterol rising to 0.8 compared with 0.2 in native LDL. When the ratio exceeded 0.6, both plasmin/CEH-LDL and MMP/CEH-LDL fused into larger particles. In parallel, they gained C-reactive protein-dependent complement-activating capacity. E-LDL produced with any protease/CEH combination was efficiently taken up by human macrophages, whereby marked induction of MMP-2 expression by E-LDL was observed. These in vitro findings had their in vivo correlates: urokinase-type plasminogen activator, MMP-2, and MMP-9 were detectable in both early and advanced human atherosclerotic lesions in colocalization with E-LDL.

Background-Treatment of low-density lipoprotein (LDL) with a protease and cholesterolesterase transforms the lipoprotein to an entity that resembles lipoprotein particles in atherosclerotic lesions, which have a high content of free cholesterol, reflecting extensive de-esterification in the intima. Because de-esterification would occur beneath the endothelium, we examined the effects of enzymatically modified LDL (E-LDL) on cultured endothelial cells. Methods and Results-Incubation of endothelial cells with E-LDL provoked selective accumulation of interleukin (IL)-8 mRNA and production of the cytokine. Chemical analyses and depletion experiments indicated that the effect was caused by the presence of free fatty acids in the altered lipoprotein. Reconstitution studies demonstrated that the oleic and linoleic acids associated with E-LDL are particularly effective IL-8 inducers. The effects of E-LDL on endothelial cells could be abrogated with albumin. Conclusion-IL-8 is required for rolling monocytes to adhere firmly to the endothelium; thus, the findings reveal a link between subendothelial entrapment of LDL, cleavage of cholesterol esters, and monocyte recruitment into the lesion.

Outbreaks of H5N1 avian influenza show strong seasonality. It is not clear where the source of virus originates from in each new outbreak season. This study sought to understand the nature of viral resurgence in recent outbreak seasons in Thailand, where the epidemic is relatively well controlled. In such a situation, indigenous viruses surviving the inter-outbreak season would have to pass through a bottleneck. In order to look for evidence of the bottleneck effect, viral genome sequences from recent outbreaks in the country were analysed. H5N1 avian influenza viruses were isolated from six outbreaks in the rainy season and winter of 2007 through to early 2008. Most of the outbreaks were in the Yom-Nan River basin in the southern part of the northern region of the country. Sequences of these viral isolates were identified as clade 1, genotype Z, similar to viruses from previous years in the central region of the country. The sequences clustered into two groups, one of which was closely related to viruses isolated from the same area in July 2006. These analyses indicated that there was a strong bottleneck effect on the virus population and that only a few lineages remained in the area. In addition, evidence of reassortment among these viruses was found. These indicated re-emergence of viruses from a small pool of indigenous sources that had been silently perpetuated over the dry summer months. Therefore, an approach to eradicate H5N1 avian influenza from the area by eliminating these local reservoirs may be feasible and should be seriously considered.

Oomycetes are unique eukaryotic microorganisms that share a mycelial morphology with fungi. Many oomycetes are pathogenic to plants, and a more limited number are pathogenic to animals. Pythium insidiosum is the only oomycete that is capable of infecting both humans and animals, and causes a life-threatening infectious disease, called “pythiosis”. In the majority of pythiosis patients life-long handicaps result from the inevitable radical excision of infected organs, and many die from advanced infection. Better understanding P. insidiosum pathogenesis at molecular levels could lead to new forms of treatment. Genetic and genomic information is lacking for P. insidiosum, so we have undertaken an expressed sequence tag (EST) study, and report on the first dataset of 486 ESTs, assembled into 217 unigenes. Of these, 144 had significant sequence similarity with known genes, including 47 with ribosomal protein homology. Potential virulence factors included genes involved in antioxidation, thermal adaptation, immunomodulation, and iron and sterol binding. Effectors resembling pathogenicity factors of plant-pathogenic oomycetes were also discovered, such as, a CBEL-like protein (possible involvement in host cell adhesion and hemagglutination), a putative RXLR effector (possibly involved in host cell modulation) and elicitin-like (ELL) proteins. Phylogenetic analysis mapped P. insidiosum ELLs to several novel clades of oomycete elicitins (ELIs), and homology modeling predicted that P. insidiosum ELLs should bind sterols. Most of the P. insidiosum ESTs showed homology to sequences in the genome or EST databases of other oomycetes, but one putative gene, with unknown function, was found to be unique to P. insidiosum. The EST dataset reported here represents the first steps in identifying genes of P. insidiosum and beginning transcriptome analysis. This genetic information will facilitate understanding of pathogenic mechanisms of this devastating pathogen.► Pythium insidiosum is the only oomycete that causes a fatal infectious disease in humans. ► Genetic and genomic information is lacking for P. insidiosum. ► We report on the first dataset of ∼500 P. insidiosum ESTs. ► Several potential virulence factors, including effectors, were discovered. ► ESTs will facilitate understanding of pathogenic mechanisms of P. insidiosum.

The recombinant envelope protein (gp120) of the human immunodeficiency virus type 1 (HIV-1) CRF01_AE env gene isolated from the corresponding blood (rgp120-F36PC) and genital fluid (rgp120-F36VC) specimens obtained from HIV infected individuals was successfully produced in both prokaryote and eukaryote cells. The yields of HIV-1 recombinant envelope proteins rgp120-F36PC and rgp120-F36VC produced in E. coli and in mammalian cells were 1.0 and 1.2, and 0.3 and 0.5 mg/ml, respectively. Antibody responses in mice immunized with rgp120-F36VC protein were not significantly higher than those with rgp120-F36PC protein. The level of antibody response in mice immunized with V3 deleted recombinant gp120 proteins from rgp120-F36VC and rgp120-F36PC was not significantly different from wild type rgp120 proteins. βstrands at the tip of the V3 loop of the HIV-1 envelope protein were predicted for the wild type genital fluid isolate but not for the wild type blood isolate. The replication capacity of both F36PC and F36VC was quite efficient. The infectivity assay of the epithelial cell line for pNL4-3/gp120F36VC was better than for pNL4-3/gp120F36PC. The extra β-strands in the V3 loop may be involved in cell tropism.

Influenza B virus, which causes acute respiratory infections, has increased in prevalence in recent years. Based on the nucleotide sequence of the hemagglutinin (HA) gene, influenza B virus can be divided into two lineages, Victoria and Yamagata, that co-circulate during the influenza season. However, analysis of the potential association between the clinical and virological characteristic and the lineage of influenza B viruses isolated in Thailand was lacking. To investigate influenza B virus genetically and determine its neuraminidase (NA) inhibitor susceptibility phenotype, a total of 6920 nasopharyngeal-wash samples were collected from patients with influenza-like illness between the years 2011 and 2014 and were screened for influenza B virus by real-time PCR. Of these samples, 3.1% (216/6920) were confirmed to contain influenza B viruses, and 110 of these influenza viruses were randomly selected for nucleotide sequence analysis of the HA and NA genes. Phylogenetic analysis of the HA sequences showed clustering into various clades: Yamagata clade 3 (11/110, 10%), Yamagata clade 2 (71/110, 64.5%), and Victoria clade 1 (28/110, 25.5%). The analysis of clinical characteristic demonstrated that the Victoria lineage was significantly associated with the duration of hospitalization, number of deceased cases, pneumonia, secondary bacterial infection and underlying disease. When combined with phylogenetic analysis of the NA sequences, four samples showed viruses with reassortant sequences between the Victoria and Yamagata lineages. Statistical analysis of the clinical outcomes and demographic data for the reassortant strains did not differ from those of the other strains in circulation. Oseltamivir-resistant influenza B viruses were not detected. Our findings indicated the co-circulation of the Victoria and Yamagata lineages over the past four cold seasons in Bangkok. We also demonstrated differences in the clinical symptoms between these lineages.

In this manuscript, we discuss relevant socioeconomic factors for developing and implementing sensor analytic point solutions (SNAPS) as point-of-care tools to serve impoverished communities. The distinct economic, environmental, cultural, and ethical paradigms that affect economically disadvantaged users add complexity to the process of technology development and deployment beyond the science and engineering issues. We begin by contextualizing the environmental burden of disease in select lowincome regions around the world, including environmental hazards at work, home, and the broader community environment, where SNAPS may be helpful in the prevention and mitigation of human exposure to harmful biological vectors and chemical agents. We offer examples of SNAPS designed for economically disadvantaged users, specifically for supporting decision-making in cases of tuberculosis (TB) infection and mercury exposure. We follow-up by discussing the economic challenges that are involved in the phased implementation of diagnostic tools in low-income markets and describe a micropayment-based systems-as-a-service approach (pay-a-penny-per-use-PAPPU), which may be catalytic for the adoption of low-end, low-margin, low-research, and the development SNAPS. Finally, we provide some insights into the social and ethical considerations for the assimilation of SNAPS to improve health outcomes in marginalized communities.

Antimicrobial resistance (AMR) is one of the top 10 global public health threats facing humanity, especially in low-resource settings, and requires an interdisciplinary response across academia, government, countries, and societies. If unchecked, AMR will hamper progress towards reaching the United Nations Sustainable Development Goals (SDGs), including ending poverty and hunger, promoting healthy lives and well-being, and achieving sustained economic growth. There are many global initiatives to curb the effects of AMR, but significant gaps remain. New ways of thinking and operating in the context of the SDGs are essential to making progress. In this entry, we define the next generation of the AMR research network, its composition, and strategic activities that can help mitigate the threats due to AMR at the local, regional, and global levels. This is supported by a review of recent literature and bibliometric and network analyses to examine the current and future state of AMR resea...

Objective: To evaluate the efficacy and safety of poly-herbal formula Sahatsatara (SHT) in pain reduction in acute low back pain (LBP) patients. Methods: Twenty-nine patients aged 18-65 years with a history of moderate to severe acute LBP ≤3-day (score ≥4 on a 0-10 numeric rating scale [NRS]) were enrolled and randomized to receive an ibuprofen (400 mg after meals three times daily) or SHT (1,350 mg before meals three times daily) for 7 days. The non-inferiority trial margin was set at ±10 percentage points. The outcomes were measured on pain intensity on the 0-10 NRS, disability on the Thai version of the Oswestry disability index [ODI], total analgesic consumption, patient satisfaction, and safety. Results: Fourteen patients and 15 patients were randomly allocated to ibuprofen and SHT groups, respectively. The mean difference in pain intensity and disability between the two groups at day 7 adjusted according to baseline was within ±1 for pain (-0.3; 95% CI, -1.48 to 0.96) and ±10%...

The objective is to evaluate the efficacy of galantamine when a slow titration regimen is employed in Thai Alzheimer's disease (AD) patients with or without cerebrovascular disease and vascular dementia (VaD).A 6-month, multicentre, open-label, uncontrolled trial was undertaken in 75 AD patients. Eligible patients received an initial galantamine dose of 8 mg/day and escalated over 5–8 weeks to maintenance doses of 16 or 24 mg/day. Primary efficacy measures were AD Assessment Scale-cognitive subscale (ADAS-cog) and the Clinician's Interview-Based Impression of Change-Plus version (CIBIC-plus). The Behavioural Pathology in AD Rating Scale (BEHAVE AD), the AD Cooperative Study Activities of Daily Living Inventory and Pittsburgh Sleep Quality Index were the secondary efficacy variables. Analyses were based on the intent-to-treat population.Treatment with galantamine showed significant improvement in cognition on the ADAS-cog and CIBIC-plus at month 6. Galantamine showed favourable effects on activities of daily living. Behavioural symptoms and sleep quality were also significantly improved (p < 0.05). Galantamine was well tolerated. The adverse events were mild-to-moderate intensity. The most frequent adverse events commonly reported were nausea (16.4%), dizziness (9.6%) and vomiting (6.8%).The results of this study may be consistent with galantamine being an effective and safe treatment for mild-to-moderate AD patients with or without cerebrovascular disease and VaD. Flexible dose escalation of galantamine was well tolerated. The daily maintenance dose of galantamine was 16 mg/day, followed by a back up dose of 24 mg/day.

The objective of this presentation is to evaluate the economic burden of BPSD within a prospective study about the economic burden of Alzheimer's disease (AD) in seventy one communtiy dwelling AD patients (mean age=76.4, mean MMSE= 12.0). The intervxws covered information about the number of care-giving hours and expenditures (i.e. medication, paid help and daycare) invested in caring for the patient.ln order to determine the amount of time invtxted to manage BPSD, caregiverc were asked to report the number of hours spent in a typical week dealing with BPSD (managing aggression, pacing, attempts to leave the house under inappropriate circumstances, or comforting a hallucinating, depressed or extremely anxious patient). We calculated the formal and informal cost of AD in Israel. The informal cost was valued as the multiplication of the hours of care by the wage of a worker who would be hired in the absence of a caregiver. The overall annual cobt of AD in Israel was $13,225 of which $9,300 in informal cost. It was found that family members provided a total of 270 care-giving hours per month per patient, out of which 28% (75 hours) were related directly to the management of BPSD, 19% to assistance with ADL, 36% to help with IADL and 17% to supervision. Hence at an hourly wage of $3 which is the cost of an unskilled nursing aid, the Informal annual cost for BPSD management comes up to approximately $2,700 ($3 X 75 hours X 12 months). The formal cost of AD, i.e. out of pocket payment for medications, daycare and paid help services amounted to $3,925 per year. Assuming that the needs of the patient are the same regardless of who providea the care (paid help or informal caregiver), it can be hypothesized that the same 28% of the care-giving effort is devoted to BPSD. Hence, the cost of BPSD in formal care is $945 per year (28% of $3,375, which is the component of paid help in formal cost) The annual out of pocket for psychotropic medication is approximately $360. Thus, the annual formal cost of BPSD IS approximately $1305 (945+360(36% of the total formal coat). Conclusion: The total annual cost of a non-institutionalized AD patient in Israel is $13,225 ($9,300 + $3.92.5). Approximately 30% ($4,000) of this cost is invested in the direct management of BPSD. This cost doe\ not include visits to physicians, hospitahzation and part of the payment for psychotropic medication, which are related to BPSD but covered hi the \tandard compulsory ~n\urance.