Leptin-induced mitochondrial fusion mediates hepatic lipid accumulation.

BACKGROUND: Leptin alleviates metabolic conditions such as insulin resistance and obesity, although the precise mechanism of action is unclear. Mitochondrial fusion/fission states affect energy balance, but the association between mitochondrial fusion and lipid metabolism is also unknown. The aim of this study was to determine whether mitochondrial fusion/fission state regulates lipid accumulation and to understand the role of leptin in mitochondrial function and its mechanism of action in metabolic regulation. METHODS: Primary mouse hepatocytes were isolated from C57BL/6J mice and treated with leptin (25 ng ml(-1)) for 3 days before determinations of mitochondrial morphology and fatty acid accumulation. Hyperglycemia in C57BL/6J mice was induced by providing a 30% fructose-rich diet (FRD) for 6 months, followed by intraperitoneal injections of leptin (1 mg kg(-1) per body weight) for 6 weeks (twice per week). RESULTS: Leptin triggered mitochondrial fusion and alleviated high glucose-induced fatty acid accumulation in primary hepatocytes by promoting mitochondrial fusion-associated transcription factor peroxisome proliferative-activated receptor-α and co-activator peroxisome proliferative-activated receptor-γ co-activator (PGC)-1α. In turn, these activate the fusion protein mitofusin 1 (Mfn-1). RNA silencing of Mfn-1 or PGC-1 blocked the inhibitory effect of leptin. Leptin treatment also elevated liver Mfn-1 and PGC-1α and improved lipid profiles in FRD mice. CONCLUSIONS: Mitochondrial fusion has a critical role in alleviating hepatic fatty acid accumulation. Leptin switches mitochondrial morphology via a PGC-1α-dependent pathway to improve hyperlipidemia.

TNIP1, SLC15A4, ETS1, RasGRP3 and IKZF1 are associated with clinical features of systemic lupus erythematosus in a Chinese Han population.

Systemic lupus erythematosus (SLE) is an autoimmune disease with heterogeneous clinical manifestations influenced by genetic and environmental factors. Five novel susceptibility genes (TNIP1, SLC15A4, ETS1, RasGRP3 and IKZF1) for SLE have been identified in a recent genome-wide association study of a Chinese Han population. This study investigated their relationships with disease subphenotypes, including renal nephritis, photosensitivity, antinuclear antibody (ANA), age at diagnosis, malar rash, discoid rash, immunological disorder, oral ulcer, hematological disorder, neurological disorder, serositis, arthritis and vasculitis. Significant associations were found for the single nucleotide polymorphism rs10036748 of TNIP1 with photosensitivity (odds ratio (OR) = 0.87, p = 0.01) and vasculitis (OR = 1.18, p = 0.04); rs10847697 of SLC15A4 with discoid rash (OR = 1.18, p = 0.02); rs6590330 of ETS1 with SLE of age at diagnosis <20 years (OR = 1.24, p = 8.91 x 10(-5)); rs13385731 of RasGRP3 with malar rash (OR = 1.20, p = 0.01), discoid rash (OR = 0.78, p = 0.02) and ANA (OR = 0.72, p = 0.004); rs4917014 of IKZF1 with renal nephritis (OR = 1.13, p = 0.02) and malar rash (OR = 0.83, p = 0.00038), respectively. The study suggested that these susceptibility genes might not only play important roles in the development of SLE, but also contribute to the complex phenotypes of SLE.

Red mold rice prevents the development of obesity, dyslipidemia and hyperinsulinemia induced by high-fat diet.

OBJECTIVE: To investigate the influences of red mold rice (RMR) on obesity and related metabolic abnormalities. METHODS AND RESULTS: The 3T3-L1 cell line was used to examine the effects of RMR extracts on preadipocytes and on mature adipocytes. Both water and ethanol extracts of RMR had inhibitory effects on 3T3-L1 preadipocyte proliferation and differentiation. Water extracts of RMR enhanced the lipolysis activity in mature adipocytes, which negatively correlated with the triglyceride content within cells. RMR treatment did not affect heparin-releasable lipoprotein lipase activity in mature adipocytes. Furthermore, animal studies were carried out to explore the antiobesity effects of RMR. The control group of male Wistar rats were fed regular laboratory feed, whereas the other groups were fed the high-fat (HF) diet supplemented with lovastatin, rice or RMR (0.4 and 2%, w w(-1)). The relative caloric intakes of the control and HF groups were 3.34 and 4.85 kcal g(-1), respectively. After 6 weeks, rats treated with RMR at the 0.4 and 2% doses had lower weight gain and less fat pads mass accompanied with smaller fat cells than did the HF-diet rats. These effects probably resulted from an increase in the lipolysis activity of adipose tissue and a reduction in food/energy consumption. On the other hand, the RMR supplement significantly reduced serum total cholesterol, serum low-density lipoprotein (LDL) cholesterol, the ratio of LDL to high-density lipoprotein (HDL) cholesterol and serum insulin in the HF group. Moreover, the 2% RMR treatment significantly increased serum HDL cholesterol. CONCLUSION: This study reveals for the first time that RMR can prevent body fat accumulation and improve dyslipidemia. The antiobesity effects of RMR mainly derive from the lipolytic activity and mild antiappetite potency of RMR. In addition, extracts of RMR suppressed the proliferation and differentiation in 3T3-L1 preadipocytes, which might have contributed to the inhibition of new adipocyte formation or hyperplasia in adipose tissue.

Screening procedure from cattle feces and the prevalence of Escherichia coli O157:H7 in Taiwan dairy cattle.

A procedure has been established for screening Escherichia coli O157:H7 from bovine feces. It consists of four steps: enrichment, selective culture, phenotyping and genotyping. Modified trypticase soy broth (mTSB) containing 20 microg/mL of novobiocin was used for the enrichment step. The selective culture step was done using sorbitol MacConkey agar containing 0.05 microg/mL of cefixime and 2.5 microg/mL of potassium tellurite. The phenotyping steps include: species confirmation as E. coli, serotyping to O157 and H7, and the detection of verocytotoxin (VT) production. Genotyping was made up of confirmation of the presence of the O157 antigen (rfb), the H7 antigen (ficC), the adherence factor (eaeA), hemolysin (hlyA), and VT production (vt I and vt II) by polymerase chain reaction. mTSB gave better enrichment for E. coli O157 than gram-negative (GN) broth. The detection limit for this screening procedure was 0.85 +/- 0.5 cfu/g. Using the screening procedure described above, E. coli O157 was found in four out of 3062 (0.13%) fresh bovine fecal specimens, and in two out of 78 (2.56%) dairy herds in Taiwan. Three out of the four detected strains were vt II producing E. coli O157:H7, while the other was E. coli O157:H7 which lacked VT-producing ability.

Molecular typing of Vibrio parahaemolyticus isolates, obtained from patients involved in food poisoning outbreaks in Taiwan, by random amplified polymorphic DNA analysis.

Vibrio parahaemolyticus is one of the most important food-borne pathogens in Taiwan, Japan, and other countries with long coastlines. This paper reports on the development of a new random amplified polymorphic DNA (RAPD) method for the molecular typing of this pathogen. The 10-mer primer 284 (5'-CAG GCG CAC A-3') was selected to generate polymorphic amplification profiles of the genomic DNA at an annealing temperature of 38 degrees C. A total of 308 clinical isolates of V. parahaemolyticus collected during food poisoning outbreaks in Taiwan, mostly occurring between 1993 and 1995, plus 11 environmental and clinical reference strains were analyzed by this RAPD method. A total of 41 polymorphic RAPD patterns were recognized, and these patterns were arbitrarily grouped into 16 types (A to P). Types A, B, C, D, and E were the major types, and subtypes C3, C5, E1, B1, D2, and A2 were the major patterns. The major types were phylogenetically more closely related to each other than to any of the minor types.

Prospective surveillance of children with invasive Haemophilus influenzae disease in Taiwan.

During the 16 months from January 1, 1996 through April 30, 1997, forty-three cases of invasive Haemophilus influenzae disease were identified in residents younger than 14 years of age in Taiwan. H. influenzae serotyping was performed on all isolated specimens. There were 33 (76.7%) cases of type b disease; three (7.0%), non-type b, and seven (16.3%), nontypeable cases. Among these H. influenzae type b cases, there were 18 (54.5%) male patients and 15 (45.5%) female patients. With regard to age-distribution, nine (27.3%) patients were aged 2 to 5 years; nine (27.3%), between 1 to 2 years; fifteen (45.5%), younger than 1; and none were younger than 3 months old. Demographical study indicated that 13 patients (39.4%) located in northern Taiwan; 5 (15.2%), central Taiwan; 12 (36.4%), southern Taiwan; 3 (9.0%) from eastern Taiwan. Among the 33 H. influenzae type b cases, twenty-five (75.8%) patients had meningitis and 8 patients had other disease entities, i.e. pneumonia in 4 patients, bacteremia in 3, and cellulitis in 1. In terms of prognosis, three (9.1%) patients died, all of whom having meningitis or sepsis; 7 (21.2%) developed hydrocephalus; 2 (6.1%) had seizure disorder without hydrocephalus and 21 (63.6%) patients recovered completely without sequelae.

[Relatedness of Shigella sonnei isolates from six outbreaks in Tao-Yuan area, Taiwan].

Between November 1995 and January 1996, one larger-scale outbreak of Shigella sonnei infection occurred in the area of Tao-Yuan, Taiwan. Subsequently, five outbreaks of S. sonnei infection occurred in the same area during the period from October 1998 to February 1999. Fifty-five isolates obtained from these six outbreaks were differentiated by antimicrobial susceptibility testing, plasmid profile analysis (PPA) and pulsed-field gel electrophoresis (PFGE). They were classified into two and three types by antimicrobial susceptibility testing and PPA, respectively. However, only one pattern was observed by PFGE. The molecular epidemiology by PFGE in this study indicated that they were closely-related. These data suggest that the clinical isolates obtained during the period of January 1998 to January 1999 were probably derived from the outbreak strain of 1995.

Food-borne disease outbreaks due to bacteria in Taiwan, 1986 to 1995.

Between 1986 and 1995, 852 outbreaks of food-borne disease involving 26,173 cases and 20 deaths were reported in Taiwan. About 80% of the outbreaks occurred in the warmer months, i.e., between April and October. Of the 852 reported outbreaks, 555 (65%) were caused by bacterial pathogens. The three most common bacteria involved were Vibrio parahaemolyticus (35%, 197 of 555 outbreaks), Staphylococcus aureus (30%, 169 of 555 outbreaks), and Bacillus cereus (18%, 104 of 555 outbreaks).

[Diagnosis of Yersinia pestis].

There is no plaque case report in Taiwan since 1952. However, it is necessary to set up a laboratory system to investigate the distribution of Yersinia pestis in the natural environment to implement the public policy for preventing plague. Besides the traditional methods; e.g. culture, microscopic observation, biochemical characteristics, anti-F1 antigen detection by slide agglutination, immunofluorescence, and phage lytic assay, PCR was used as rapid screening test in our study. These laboratory methods were used to examine whether the flea samples harvested in King-Men island carry Y. pestis. The results showed that the flea index per mouse was high but no Y. pestis was detected in the fleas.

[Pathogenic strains of Escherichia coli in Taiwan].

From July 1994 through June 1996, 28 strains of Escherichia coli were isolated from 1,260 patients with acute diarrhea. These strains were further differentiated with serotypes and virulence factors. Enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), enterohemorrhagic E. coli (EHEC), and enteroinvasive E. coli (EIEC) were accounted for 53.6 (15 of 28 strains), 28.6 (8 of 28), 10.7 (3 of 28) and 7.1% (2 of 28), respectively. Therefore, ETEC and EPEC are playing an important role in food-borne illness in Taiwan. Escherichia coli O157:H7, a new emerging pathogen of food-borne disease, has not been isolated in this study.

[K-serotype analyses of Vibrio parahaemolyticus isolated in northern Taiwan, 1983 through 1993].

From 1983 through 1993, 786 strains of Vibrio parahaemolyticus were collected from food-borne disease outbreaks and sporadic cases of diarrheal illness in northern Taiwan, involving 42 K-serotypes. Five top leading serotypes were K8 (36.8%), K15 (10.8%), K12 (8.7%), K56 (7.9%) and K63 (4.7%). However, a variation of K-serotypes was found during this study period. From 112 food-borne outbreaks associated with this microorganism, only 54 (48.2%) outbreaks were caused by a single serotype, while 58 (51.8%) were caused by multiple K-serotypes. Numbers of outbreaks caused by two, three and more than three K-serotypes were 29 (26%), 16 (14.2%), and 13 (11.6%), respectively. In a special outbreak, eight K-serotypes was found. Outbreaks caused by party caterers were most frequently associated with multiple K-serotypes.

[Serotype distribution and antimicrobial susceptibility of group A streptococci (Streptococcus pyogenes) isolated in Taiwan].

T-protein serotypes and antimicrobial susceptibility of a total of 139 group A streptococci (GAS) strains isolated in Taiwan area in 1993 and during the outbreak of scarlet fever in 1994 were analyzed. All strains were T-typable, and T12 (42.46%) and T4 (38.85%) were the dominant T types. According to the results of analysis of antimicrobial susceptibility, all GAS strains were divided into 9 resistotypes, A (all susceptible), B (resistant to tetracycline), C (resistant to erythromycin and tetracycline), D (resistant to chloramphenicol and tetracycline), E (resistant to chloramphenicol and clindamycin), F (resistant to chloramphenicol, clindamycin and tetracycline), G (resistant to clindamycin, erythromycin and tetracycline), H (resistant to chloramphenicol, clindamycin, erythromycin and tetracycline), and I (resistant to chloramphenicol, clindamycin, erythromycin, tetracycline and vancomycin). Type B (37.42%) was the dominant type. Type A (25.91%), and type H (26.63%) also appered with high incidence. Most of strains isolated from Mid-Taiwan were type H. Only one strain, that was isolated in I-lan, was resistant to vancomycin, in addition to resistant to chloramphenicol, clindamycin, erythromycin, and tetracycline. All strains were susceptible to penicillin G, ampicillin, and ceftriaxone. Some strains were resistant to chloramphenicol (32.38%), clindamycin (30.22%), erythromycin (31.66%), tetracycline (73.39%), and vancomycin (0.70%). During the outbreak of scarlet fever in 1994, the dominant T types of strains isolated in North-Taiwan and Mid-Taiwan were T4 and T12, respectively, and the major resistotypes of those strains were B and H types, respectively. These clues suggested that the outbreaks occurring in North-Taiwan and Mid-Taiwan may have no epidemiological linkage between each other.

Legionella pneumophila infection in Taiwan: a preliminary report.

Legionella pneumophila infection has been recognized as an important cause of pneumonia. However, epidemiologic baseline data about the occurrence of legionella-related pneumonia and the distribution of legionellae in aquatic environments are still not available in Taiwan. A survey conducted by the National Institute of Preventive Medicine found that 8.6% of 487 pneumonia patients studied were positive for the L. pneumophila serologic test (indirect fluorescent test, IFA) and 32.0% of the 25 water samples were positive for isolation of legionellae. In conclusion, this survey supported the existence of Legionnaires' disease in Taiwan, indicated a preliminary prevalence of L. pneumophila antibodies among pneumonia patients and identified the prevalence of legionellae in water environments.

Subspecies typing of Vibrio parahaemolyticus by pulsed-field gel electrophoresis.

Vibrio parahaemolyticus is one of the most important food-borne pathogens in Taiwan, Japan, and other costal regions. We report on the development of a pulsed-field gel electrophoresis (PFGE) method for the molecular typing of this pathogen. Genomic DNA was digested with SfiI, and the fragments were resolved on 1% agarose with a contour-clamped homogeneous electric field apparatus set at 190 V and a pulse time of 3 to 80 s. A total of 130 selected isolates obtained from outbreaks during 1993 and 1994 on Taiwan were also characterized by this PFGE method. These isolates were grouped into 14 PFGE types which consisted on one to six patterns, and a total of 39 patterns were identified. Most of these domestic clinical isolates could be clustered into several major types (types A, B, C, and G). These major types showed relatively low degrees of similarity to several foreign strains and other domestic but environmental strains. Strain CCRC12863, which originated from Japan, was close to the group consisting of F, G, and H PFGE types, suggesting a clonal relationship between this Japanese strain and other domestic isolates.

Food-borne disease outbreaks in Taiwan, 1994.

In 1994, 102 outbreaks of food-borne disease involving 4,726 cases were reported to the Taiwan Department of Health. This is the highest number of outbreaks and cases in recent years in Taiwan. Of these outbreaks, 72.5% (74/102) were caused by bacterial pathogens, with Vibrio parahaemolyticus responsible for 56.7% (42/74), Staphylococcus aureus 20.3% (15/74), Bacillus cereus 14.9% (11/74) and Salmonella spp other than S. typhi and S. paratyphi 8.1% (6/74). V. parahaemolyticus has been a leading cause of problems in Taiwan for many years. Contamination of seafood with this organism has been reported frequently, particularly in the warmer months. In 1994, small outbreaks (fewer than 5 cases) and large outbreaks (more than 50 cases) represented 31.4% (32/102) and 12.7% (13/102), respectively, of the total. The median outbreak size was 10 cases. A high proportion (54%, 7/13) of the large outbreaks was associated with commercial lunch-boxes supplied to elementary and junior high schools. Health education to improve food sanitation and supervision of food sanitation practices need to be strengthened.

[Comparison of immunoassay kits for detection of staphylococcal enterotoxins produced by Staphylococcus aureus].

UNLABELLED: Four commercial kits, SET-RPLA, RIDASCREEN, SET-EIA, and TECRA, were compared for the efficiency of detecting staphylococcal enterotoxins (SE). There was no non-specific reaction for detection of SE produced by 21 Staphylococcus aureus strains from 5 outbreaks of food poisoning using SET-RPLA, SET-EIA and TECRA kits. The background of the results of RIDASCREEN kit was high and non-specific reactions were present in some strains. IN CONCLUSION: (i) TECRA kit is suggested to be used for screening SE producing strains; (ii) SET-RPLA and RIDASCREEN kits are suitable for epidemiological investigation of SE types, but the lack of ability for detecting SEE, long time required for testing with SET-RPLA kit and high background when using RIDASCREEN kit must be overcome; and (iii) because of the complicated test procedures and the lack of ability for detecting SEE, the practicality of SET-EIA kit in screening and epidemiological research purposes is low.

Deodorization of laboratory animal facilities by ozone.

Deodorizing effect of ozone was investigated comparing two types of compact ozonizing apparatus made on an experimental basis. The concentrations of ammonia and trimethylamine were examined as an indicator for deodorizing effect of ozone in animal rooms of rats and guinea pigs at laboratory animal facilities of three different universities. Both of the ozonizing apparatus were able to remove ammonia and trimethylamine in animal rooms, with no significant difference in the performance of the two apparatus.

Latex agglutination test for detection of tetanus antitoxins.

A rapid and easy method of slide agglutination test for the detection of human tetanus antitoxins was developed in this study. Testing reagents were prepared from carboxylated polystyrene latex particles with tetanus toxin by soluble carbodiimide. The test was performed on a glass slide with a drop of test sample and a drop of testing reagent. The agglutination reaction was usually completed within five minutes. Sensitivity of this test for tetanus antitoxins can be reached at 0.125 IU/ml. Therefore, the latex agglutination test can be used to determine the immune status of a patient in an emergency.

A preliminary report on Borrelia burgdorferi infection in the Taiwan area.

A total of 273 serum specimens from different areas and sources were tested against Borrelia burgdorferi antigens by enzyme-linked immunosorbent assay method. Positive rates of serological reactions were 3% and 58% for healthy persons and syphilis patients, respectively. Obviously, there was a lot of cross-reaction in the venereal disease group. Meanwhile, positive rates were 3% and 13% in the sera collected from Taiwan and Orchid Island, respectively. This difference may reflect a less developed environment in the latter. Since reported cases of Lyme disease in Taiwan are rare, serologic tests are usually adapted for rapid diagnosis in common laboratories. As for disease confirmation, clinical observations, epidemiological data and exposure in an endemic area must also be considered.