Using the lactate-to-albumin ratio to predict mortality in patients with sepsis or septic shock: a systematic review and meta-analysis.

OBJECTIVE: This study aimed to investigate whether the lactate-to-albumin ratio (LAR) can predict mortality in patients with sepsis or septic shock. PATIENTS AND METHODS: A systematic search of the PubMed, EMBASE, Web of Science, and Google Scholar databases was conducted on December 16, 2021, for relevant articles that provided the predictive performance of LAR for mortality in patients with sepsis or septic shock. RESULTS: Eight studies encompassing a total of 4,723 patients were included in this paper. The pooled sensitivity, specificity, and diagnostic odds ratio of the LAR for predicting mortality were 0.71 (95% confidence interval [CI]: 0.54-0.84), 0.68 (95% CI: 0.58-0.76) and 5.23 (95% CI: 2.62-10.45), respectively. The area under the summary receiver operating characteristic curve was 0.74 (95% CI: 0.70-0.78). CONCLUSIONS: The current evidence suggests that LAR is moderately predictive of mortality among patients with sepsis or septic shock and may be beneficial to identify high-risk patients.

Neutrophil-to-lymphocyte ratio for the diagnosis of pediatric acute appendicitis: a systematic review and meta-analysis.

OBJECTIVE: Acute appendicitis (AA) is one of the most common surgical emergencies and causes of acute abdominal pain in the pediatric population. However, it can be difficult to diagnose in children. We aimed to provide updated evidence on the diagnostic utility of the neutrophil-to-lymphocyte ratio (NLR) for AA, along with other conventional biomarkers, in pediatric patients. MATERIALS AND METHODS: We searched the PubMed, Embase, Cochrane Library, and Web of Science databases for eligible articles published up to May 16, 2021. RESULTS: We included 19 studies comprising a total of 5,974 pediatric cases. The overall sensitivity and specificity of the NLR were 0.82 (95% confidence interval [CI]: 0.79-0.85) and 0.76 (95% CI: 0.69-0.81), respectively. The overall diagnostic odds ratio was 14.34 (95% CI: 9.05-22.73). The area under the summary receiver operating characteristic curve was 0.86 (95% CI: 0.83-0.89). The pooled sensitivity and specificity of other biomarkers were as follows: 0.79 (95% CI: 0.71-0.86) and 0.66 (95% CI: 0.54-0.77) for the white blood cell count, 0.73 (95% CI: 0.69-0.77) and 0.68 (95% CI: 0.55-0.79) for the C-reactive protein level, 0.75 (95% CI: 0.65-0.82) and 0.78 (95% CI: 0.72-0.83) for the absolute neutrophil count, and 0.83 (95% CI: 0.79-0.87) and 0.68 (95% CI: 0.53-0.80) for the neutrophil percentage, respectively. CONCLUSIONS: The NLR has moderate predictive power for AA and can be used as a simple, auxiliary tool for diagnosis. NLR can also help clinicians decide whether to perform imaging testing when the clinical symptoms or physical examination findings are vague.

Airborne particles in indoor environment of homes, schools, offices and aged care facilities: The main routes of exposure.

It has been shown that the exposure to airborne particulate matter is one of the most significant environmental risks people face. Since indoor environment is where people spend the majority of time, in order to protect against this risk, the origin of the particles needs to be understood: do they come from indoor, outdoor sources or both? Further, this question needs to be answered separately for each of the PM mass/number size fractions, as they originate from different sources. Numerous studies have been conducted for specific indoor environments or under specific setting. Here our aim was to go beyond the specifics of individual studies, and to explore, based on pooled data from the literature, whether there are generalizable trends in routes of exposure at homes, schools and day cares, offices and aged care facilities. To do this, we quantified the overall 24h and occupancy weighted means of PM10, PM2.5 and PN - particle number concentration. Based on this, we developed a summary of the indoor versus outdoor origin of indoor particles and compared the means to the WHO guidelines (for PM10 and PM2.5) and to the typical levels reported for urban environments (PN). We showed that the main origins of particle metrics differ from one type of indoor environment to another. For homes, outdoor air is the main origin of PM10 and PM2.5 but PN originate from indoor sources; for schools and day cares, outdoor air is the source of PN while PM10 and PM2.5 have indoor sources; and for offices, outdoor air is the source of all three particle size fractions. While each individual building is different, leading to differences in exposure and ideally necessitating its own assessment (which is very rarely done), our findings point to the existence of generalizable trends for the main types of indoor environments where people spend time, and therefore to the type of prevention measures which need to be considered in general for these environments.

Inhibition of DNA2 nuclease as a therapeutic strategy targeting replication stress in cancer cells.

Replication stress is a characteristic feature of cancer cells, which is resulted from sustained proliferative signaling induced by activation of oncogenes or loss of tumor suppressors. In cancer cells, oncogene-induced replication stress manifests as replication-associated lesions, predominantly double-strand DNA breaks (DSBs). An essential mechanism utilized by cells to repair replication-associated DSBs is homologous recombination (HR). In order to overcome replication stress and survive, cancer cells often require enhanced HR repair capacity. Therefore, the key link between HR repair and cellular tolerance to replication-associated DSBs provides us with a mechanistic rationale for exploiting synthetic lethality between HR repair inhibition and replication stress. DNA2 nuclease is an evolutionarily conserved essential enzyme in replication and HR repair. Here we demonstrate that DNA2 is overexpressed in pancreatic cancers, one of the deadliest and more aggressive forms of human cancers, where mutations in the KRAS are present in 90-95% of cases. In addition, depletion of DNA2 significantly reduces pancreatic cancer cell survival and xenograft tumor growth, suggesting the therapeutic potential of DNA2 inhibition. Finally, we develop a robust high-throughput biochemistry assay to screen for inhibitors of the DNA2 nuclease activity. The top inhibitors were shown to be efficacious against both yeast Dna2 and human DNA2. Treatment of cancer cells with DNA2 inhibitors recapitulates phenotypes observed upon DNA2 depletion, including decreased DNA double strand break end resection and attenuation of HR repair. Similar to genetic ablation of DNA2, chemical inhibition of DNA2 selectively attenuates the growth of various cancer cells with oncogene-induced replication stress. Taken together, our findings open a new avenue to develop a new class of anticancer drugs by targeting druggable nuclease DNA2. We propose DNA2 inhibition as new strategy in cancer therapy by targeting replication stress, a molecular property of cancer cells that is acquired as a result of oncogene activation instead of targeting currently undruggable oncoprotein itself such as KRAS.

Increased propionate concentration in Lactobacillus mucosae-fermented wet brewers grains and during in vitro rumen fermentation.

AIMS: This study was conducted to isolate and identify propionate-producing bacteria that can be used as an inoculum in improving wet brewers grains and rumen fermentation via increasing propionate concentration. METHODS AND RESULTS: A strain of Lactobacillus that exhibits high levels of propionate production was identified and characterized as Lactobacillus mucosae 521129 by 16S rRNA gene sequencing and phylogenetic analyses. Wet brewers grains were fermented through L. mucosae inoculation and resulted in an increase in propionate concentration. Fermented wet brewers grains were used in in vitro rumen fermentation and revealed that L. mucosae-fermented wet brewers grains produced more gas and had higher accumulations propionate and total volatile fatty acid (VFA) than the control. The fewest methanogen DNA copies were detected in L. mucosae-fermented wet brewers grains. CONCLUSION: Identified L. mucosae improved the fermentation of wet brewers grains and the in vitro rumen fermentation via increasing propionate and total VFA concentrations. SIGNIFICANCE AND IMPACT OF THE STUDY: The presented research provided the identification of L. mucosae 521129 as a propionate producer and was metabolically profiled. Furthermore, data present the putative application of this organism in improving the fermentation of wet brewers grains and in vitro rumen fermentation.

Trends in the incidence rate of device-associated infections in intensive care units after the establishment of the Korean Nosocomial Infections Surveillance System.

BACKGROUND: The effectiveness of continuous nationwide surveillance on healthcare-associated infections should be investigated in each country. AIM: To assess the rate of device-associated infections (DAIs) in intensive care units (ICUs) since the establishment of the Korean Nosocomial Infections Surveillance System (KONIS). METHODS: Nationwide data were obtained on the incidence rate of DAI in ICUs reported to KONIS by all participating hospitals. The three major DAIs were studied: ventilator-associated pneumonia (VAP), central line-associated bloodstream infection (CABSI), and catheter-associated urinary tract infection (CAUTI). The pooled and year-wise incidence rates (cases per 1000 device-days) of these DAIs were determined for the period 2006 and 2012. In addition, data from institutions that had participated in KONIS for at least three consecutive years were analysed separately. FINDINGS: The number of ICUs participating in KONIS gradually increased from 76 in 2006 to 162 in 2012. Between 2006 and 2012, the incidence rate per 1000 device-days for VAP decreased significantly from 3.48 to 1.64 (F = 11, P < 0.01), for CAUTI the rate decreased non-significantly from 1.85 to 1.26 (F = 2.02, P = 0.07), and for CABSI the rate also decreased non-significantly from 3.4 to 2.57 (F = 1.73, P = 0.12). In the 132 ICUs that had participated in KONIS for at least three consecutive years, the VAP rate significantly decreased from the first year to third year (F = 20.57, P < 0.01), but the rates of CAUTI (F = 1.06, P = 0.35) and CABSI (F = 1.39, P = 0.25) did not change significantly. CONCLUSION: The decreased incidence rate of VAP in ICUs in Korea might be associated with the continuous prospective surveillance provided by KONIS.

Nanoparticle formation in a chemical storage room as a new incidental nanoaerosol source at a nanomaterial workplace.

Chemical storage rooms located near engineered nanomaterials (ENMs) workplaces can be a significant source of unintentional nanoaerosol generation. A new incidental nanoparticle source was identified and characterized in a chemical storage room located at an ENMs workplace. Stationary and mobile measurements using on-line instruments and chemical analysis of volatile organic compounds (VOCs) were carried out to identify the source. The number of nanoaerosols emitted from the chemical storage room was found to be several orders of magnitude higher than that existing in the ENMs workplace. VOC analysis showed that the accumulated precursors and oxygenated VOCs in the chemical storage room could be attributed to incidental particle formation via gas-to-particle conversion. We stress the importance of identification of the incidental nanoaerosols to allow characterization of the nanoaerosols at ENMs workplaces, and to estimate additional nanoaerosols exposure, which was previously unknown. Hazardous chemical substances in the workplace have been regulated in many countries; however, most of the regulations are focused on gas-phase or liquid-phase substances. The present study emphasizes the importance of secondary pollutants in particulate form that can be generated from the gas or liquid phase of hazardous chemical substances.

Indoor aerosols: from personal exposure to risk assessment.

Motivated by growing considerations of the scale, severity, and risks associated with human exposure to indoor particulate matter, this work reviewed existing literature to: (i) identify state-of-the-art experimental techniques used for personal exposure assessment; (ii) compare exposure levels reported for domestic/school settings in different countries (excluding exposure to environmental tobacco smoke and particulate matter from biomass cooking in developing countries); (iii) assess the contribution of outdoor background vs indoor sources to personal exposure; and (iv) examine scientific understanding of the risks posed by personal exposure to indoor aerosols. Limited studies assessing integrated daily residential exposure to just one particle size fraction, ultrafine particles, show that the contribution of indoor sources ranged from 19% to 76%. This indicates a strong dependence on resident activities, source events and site specificity, and highlights the importance of indoor sources for total personal exposure. Further, it was assessed that 10-30% of the total burden of disease from particulate matter exposure was due to indoor-generated particles, signifying that indoor environments are likely to be a dominant environmental factor affecting human health. However, due to challenges associated with conducting epidemiological assessments, the role of indoor-generated particles has not been fully acknowledged, and improved exposure/risk assessment methods are still needed, together with a serious focus on exposure control.

Effect of dietary supplementation of Lactobacillus-fermented Artemisia princeps on growth performance, meat lipid peroxidation, and intestinal microflora in Hy-line Brown male chickens.

The objective of this experiment was to investigate the effect of dietary supplementation of Lactobacillus-fermented Artemisia princeps (LFA) on growth performance, meat lipid peroxidation, and intestinal microflora in Hy-line Brown male chickens. A total of six hundred twenty-four 1-d-old Hy-Line Brown male chicks were randomly allotted to 3 dietary treatments with 4 replicated pens consisting of 52 chicks. The control diet was formulated to be adequate in energy and nutrients. Two additional diets were prepared by adding 2.5 or 5.0 g/kg of LFA to the control diet. The experimental diets were fed on an ad libitum basis to the birds during 7 wk. Body weight gain and feed intake were recorded at 2 and 7 wk. At the end of the experiment, 2 birds from each treatment were killed by cervical dislocation and the samples for ileal content, breast, and thigh meat were collected for the determination of meat lipid peroxidation and microbial population. Results indicated that increasing inclusion level of LFA in diets improved BW gain (linear and quadratic, P < 0.05) and tended to improve feed efficiency (linear and quadratic, P < 0.10) of birds during 0 to 7 wk. Feeding the diets containing increasing amounts of LFA to birds reduced (quadratic, P < 0.05) thiobarbituric acid-reactive substance (TBARS) values in breast and thigh meat during 15 d of storage. The concentrations of Lactobacillus spp. in the ileal content of birds increased (linear and quadratic, P < 0.05), but those of Salmonella spp. tended to be decreased (quadratic, P < 0.10) as inclusion level of LFA in diets increased. These results suggest that dietary LFA may be used as a functional ingredient to improve growth performance, meat lipid stability, and intestinal health of birds.

Expression phenotype changes of EBV-transformed lymphoblastoid cell lines during long-term subculture and its clinical significance.

OBJECTIVES: The EBV-transformed lymphoblastoid cell line (LCL) is a useful resource for population-based human genetic and pharmacogenetic studies. The principal objective here was to assess expression phenotype changes during long-term subculture of LCLs, and its clinical significance. MATERIALS AND METHODS: We searched for genes that were differentially expressed in 17 LCLs at late (p161) passage compared to early passage (p4) using microarray assay, then validated them by real-time RT-PCR analysis. In addition, we estimated correlations between expression phenotypes of 20 LCL strains at early passage and 23 quantitative clinical traits from blood donors of particular LCL strains. RESULTS: Transcript sequences of 16 genes including nuclear factor-kappaB (NF-kappaB) pathway-related genes (such as PTPN13, HERC5 and miR-146a) and carcinogenesis-related genes (such as XAF1, TCL1A, PTPN13, CD38 and miR-146a) were differentially expressed (>2-fold change) in at least 15 of the 17 LCL strains. In particular, TC2N, FCRL5, CD180, CD38 and miR-146a were downregulated in all 17 of the evaluated LCL strains. In addition, we identified clinical trait-associated expression phenotypes in LCLs. CONCLUSION: Our results showed that LCLs acquired expression phenotype changes involving expression of NF-kappaB pathway- and carcinogenesis-related genes during long-term subculture. These differentially expressed genes can be considered to be a gene signature of LCL immortalization or EBV-induced carcinogenesis. Clinical trait-associated expression phenotypes should prove useful in the discovery of new candidate genes for particular traits.

Transmissible infection of human 293T cells with porcine endogenous retroviruses subgroup a from NIH-miniature pig.

In pig-to-human xenotransplantation, zoonotic infections have been an important barrier. The risk of zoonosis has been emphasized in xenotransplantation after finding that porcine endogenous retroviruses (PERVs) can infect human cells in vitro. Until now, transmissions of PERVs from PK15 cells have been studied in vitro and in vivo, but transmission of PERVs originating from miniature pigs have not been extensively reported. Peripheral blood mononuclear cells from miniature swine showed PERV transmission to human cells. In contrast, specific pathogen-free (SPF) pig islet cells showed no PERV transmission when co-incubated with 293T cells. To evaluate the risk of zoonosis with our experimental mini pigs, we tested the infectivity of PERVs from NIH-miniature pig primary ear cells for human 293T cells. As a result, all subgroups of infectious PERV virion (PERV-A, -B, and -C) were detected in the primary cell culture media. Unlike PERV-C, PERV-A and -B infected human 293T cells. Interestingly, only proviral PERV-A replicated in 293T cells to produce virions after infection. Our results suggested that a prevention study of PERV xenotransmission from experimental miniature pigs should concentrate on PERV-A control.

Improvement of pre- and post-processing environments of the dynamic two-dimensional reservoir model CE-QUAL-W2 based on GIS.

An Environmental Information System (EIS) coupled with a Geographic Information System (GIS) and water quality models is developed to improve the pre- and post-data processing function of CE-QUAL-W2. Since the accuracy of the geometric data in terms of a diverse water body has a great effect on the water quality variables such as the velocity, kinetic reactions, the horizontal and vertical momentum, to prepare the bathymetry information has been considered a difficult issue for modellers who intend to use the model. For identifying Cross Section and Profile Information (CSPI), which precisely contains hydraulic features and geographical configuration of a waterway, the automated CSPI extraction program has been developed using Avenue Language of the PC Arc/view package. The program consists of three major steps: (1) getting the digital depth map of a waterway using GIS techniques; (2) creating a CSPI data set of segments in each branch using the program for CE-QUAL-W2 bathymetry input; (3) selecting the optimal set of bathymetry input by which the calculated water volume meets the observed volume of the water body. Through those approaches, it is clear that the model simulation results in terms of water quality as well as reservoir hydraulics rely upon the accuracy of bathymetry information.

Carcinoma erysipeloides associated with anaplastic thyroid carcinoma.

Cutaneous metastases from carcinoma of the thyroid gland are rare and carcinoma erysipeloides is even rarer. We present the clinical, histological, and immunohistochemical features of inflammatory erysipeloid metastases arising from an anaplastic carcinoma of the thyroid gland. In this case the anaplastic carcinoma probably transformed from a pre-existing, long-standing papillary carcinoma of the thyroid gland. Although visible inflammation is a hallmark of many benign skin disorders, it is not commonly present in cutaneous malignant metastases. As a result, the significance of a marked inflammatory changes in association with metastatic skin disease may not be recognized. Dermatologists need to be aware of the potential for inflammatory manifestations in cutaneous metastases from a thyroid carcinoma.

Activation of p21(WAF1/Cip1) transcription through Sp1 sites by histone deacetylase inhibitor apicidin: involvement of protein kinase C.

We previously reported that apicidin, a novel histone deacetylase inhibitor, inhibited the proliferation of tumor cells via induction of p21(WAF1/Cip1). In this study, we determined the molecular mechanisms by which apicidin induced the p21(WAF1/Cip1) gene expression in HeLa cells. Apicidin induced p21(WAF1/Cip1) mRNA independent of the de novo protein synthesis and activated the p21(WAF1/Cip1) promoter through Sp1-3 site located at -82 and -77 relative to the transcription start site. This transcriptional activation appears to be mediated by protein kinase C (PKC), because calphostin C, a PKC inhibitor, significantly attenuated the activation of p21(WAF1/Cip1) promoter via Sp1 sites, which was accompanied by a marked suppression of p21(WAF1/Cip1) mRNA and protein expression induced by apicidin. Consistent with the transcriptional activation of p21(WAF1/Cip1) promoter by apicidin, apicidin treatment led to the translocation of PKCepsilon from cytosolic to particulate fraction, which was reversed by pretreatment with calphostin C, indicating the involvement of PKC in the transcriptional activation of p21(WAF1/Cip1) via Sp1 sites by apicidin. However, the PKC-mediated transcriptional activation of p21(WAF1/Cip1) by apicidin appears to be independent of the histone hyperacetylation, because apicidin-induced histone hyperacetylation was not affected by calphostin C. Furthermore, a PKC activator, phorbol 12,13-dibutyrate, alone induced the transcriptional activation of p21(WAF1/Cip1) promoter, p21(WAF1/Cip1) mRNA, and protein expression without induction of the histone hyperacetylation, suggesting that the transcriptional activation of p21(WAF1/Cip1) by apicidin might have been mediated by a mechanism other than chromatin remodeling through the histone hyperacetylation. Taken together, these results suggest that the PKC signaling pathway plays a pivotal role in the transcriptional activation of the p21(WAF1/Cip1) gene by apicidin.

Apicidin, a histone deacetylase inhibitor, inhibits proliferation of tumor cells via induction of p21WAF1/Cip1 and gelsolin.

Apicidin [cyclo(N-O-methyl-L-tryptophanyl-L-isoleucinyl-D-pipecolinyl -L-2-amino-8-oxodecanoyl)] is a fungal metabolite shown to exhibit antiparasitic activity by the inhibition of histone deacetylase (HDAC). In this study, we evaluated apicidin as a potential antiproliferative agent. Apicidin showed a broad spectrum of antiproliferative activity against various cancer cell lines, although with differential sensitivity. The antiproliferative activity of apicidin on HeLa cells was accompanied by morphological changes, cell cycle arrest at G1 phase, and accumulation of hyperacetylated histone H4 in vivo as well as inhibition of partially purified HDAC in vitro. In addition, apicidin induced selective changes in the expression of p21WAF1/Cip1 and gelsolin, which control the cell cycle and cell morphology, respectively. Consistent with increased induction of p21WAF1/Cip1, phosphorylation of Rb protein was markedly decreased, indicating the inhibition of cyclin-dependent kinases, which became bound to p21WAF1/Cip1. The effects of apicidin on cell morphology, expression of gelsolin, and HDAC1 activity in vivo and in vitro appeared to be irreversible, because withdrawal of apicidin did not reverse those effects, whereas the induction of p21WAF1/Cip1 by apicidin was reversible. Taken together, the results suggest that induction of histone hyperacetylation by apicidin is responsible for the antiproliferative activity through selective induction of genes that play important roles in the cell cycle and cell morphology.

Hydrogen peroxide activates p70(S6k) signaling pathway.

We investigated a possible role of reactive oxygen species (ROS) in p70(S6k) activation, which plays an important role in the progression of cells from G(0)/G(1) to S phase of the cell cycle by translational up-regulation of a family of mRNA transcripts that encode for components of the protein synthetic machinery. Treatment of mouse epidermal cell JB6 with H(2)O(2) generated extracellularly by glucose/glucose oxidase led to the activation of p70(S6k) and p90(Rsk) and to phosphorylation of p42(MAPK)/p44(MAPK). The activation of p70(S6k) and p90(Rsk) was dose-dependent and transient, maximal activities being in extracts treated for 15 and 30 min, respectively. Further characterization of ROS-induced activation of p70(S6k) using specific inhibitors for p70(S6k) signaling pathway, rapamycin, and wortmannin revealed that ROS acted upstream of the rapamycin-sensitive component FRAP/RAFT and wortmannin-sensitive component phosphatidylinositol 3-kinase, because both inhibitors caused the inhibition of ROS-induced p70(S6k) activity. In addition, Ca(2+) chelation also inhibited ROS-induced activation of p70(S6k), indicating that Ca(2+) is a mediator of p70(S6k) activation by ROS. However, down-regulation of 12-O-tetradecanoylphorbol-13-acetate (TPA)-responsive protein kinase C (PKC) by chronic pretreatment with TPA or a specific PKC inhibitor Ro-31-8220 did not block the activation of p70(S6k) by ROS, indicating that the activation of TPA-responsive PKC was not required for stimulation of p70(S6k) activity by H(2)O(2) in JB6 cells. Exposure of JB6 cells to platelet-derived growth factor or epidermal growth factor led to a rapid increase in H(2)O(2), phosphorylation, and activation of p70(S6k), which were antagonized by the pretreatment of catalase. Taken together, the results suggest that ROS act as a messenger in growth factor-induced p70(S6k) signaling pathway.

NO/cGMP pathway is involved in exocrine secretion from rat pancreatic acinar cells.

The enzyme responsible for the synthesis of nitric oxide (NO) from L-arginine in mammalian tissues is known as nitric oxide synthase (NOS) (EC.1.14.13.39). In the present study, the role of NO in the regulation of exocrine secretion was investigated in rat pancreatic acinar cells. Treatment of rat pancreatic acinar cells with cholecystokinin-octapeptide (CCK-OP) resulted in an increase in the arginine conversion to citrulline, the amount of NOx, the release of amylase, and the level of cGMP. Especially, CCK-OP-stimulated increase of arginine to citrulline transformation, the amount of NOx and cGMP level were completely counteracted by the inhibitor of NOS, NG-monomethyl-L-arginine (MMA), by contrast, that of amylase release was partially reduced. Furthermore, MMA-induced decrease of NOS activity and amylase release showed dose-dependent pattern. The data on the time course of CCK-OP-induced citrulline formation and cGMP rise indicate that NOS and guanylate cyclase were activated by treatment of CCK-OP. However, the mechanism of agonist-stimulated guanylate cyclase activation in acinar cells remains unknown. Therefore, activation of NOS is one of the early events in receptor-mediated cascade of reactions in pancreatic acinar cells and NO, not completely, but partially mediate pancreatic enzyme exocrine secretion.

Molecular analysis of rifampin-resistant Mycobacterium tuberculosis isolated from Korea by polymerase chain reaction-single strand conformation polymorphism sequence analysis.

OBJECTIVE: To assess the molecular mechanism of rifampin (RMP) resistance in clinical strains of Mycobacterium tuberculosis. DESIGN: The molecular nature of a part of the rpoB gene in 77 M. tuberculosis clinical strains isolated in Korea was analyzed using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and PCR-sequence analysis. RESULTS: Among 67 RMP-resistant isolates, 50 showed SSCP profiles different from that of an RMP-sensitive control strain, M. tuberculosis H37Rv, indicating the possible existence of a sequence alteration in this region of the rpoB gene, while 17 resistant isolates displayed SSCP profiles indistinguishable from that of the sensitive control strain. Subsequently, 17 clinical isolates whose SSCP profiles were difficult to distinguish from the control strain were subjected to sequence analysis. The analysis revealed that all 17 isolates did indeed contain mutations in the 81 bp region of the rpoB gene, which is associated with RMP resistance. CONCLUSION: The results from our study clearly indicate that the molecular mechanism of RMP resistance in M. tuberculosis isolates from Korea involves alterations in the rpoB gene. In addition, this study suggests that PCR-direct sequence analysis works more efficiently and accurately than PCR-SSCP analysis for rapid screening of RMP-resistant M. tuberculosis clinical isolates.