[Chylomicron retention disease caused by SAR1B gene variations in 2 cases and literatures review].

Objective: To summarize the genotype and clinical characteristics of chylomicron retention disease (CMRD) caused by secretion associated Ras related GTPase 1B (SAR1B) gene variations. Methods: Clinical data and genetic testing results of 2 children with CMRD treated at Children's Hospital of Fudan University and Jiangxi Provincial Children's Hospital from May 2022 to July 2023 were summarized. To provide an overview of the clinical and genetic characteristics of CMRD caused by SAR1B gene variations, all of the literature was searched and reviewed from China National Knowledge Infrastructure, Wanfang Data Knowledge Service Platform, China VIP database, China Biology Medicine disc and PubMed database (up to January 2024) with "chylomicron retention disease" "Anderson disease" or "Anderson syndrome" as the search terms. All relevant literatures were reviewed to summarize the clinical and genetic features of CMRD caused by SAR1B gene variations. Results: One 11-year-old boy and one 4-month-old girl with CMRD. Both patients had lipid malabsorption, failure to thrive, decreased cholesterol, elevated transaminase and creatine kinase, and Vitamin E deficiency, with homozygous variations (c.224A>G) and compound heterozygous variations (c.224A>G and c.554G>T) in SAR1B gene, respectively. Case 1 was followed up for over a month, and he still occasionally experienced lower limb muscle pain. Case 2 was followed up for more than a year, and her had caught up to normal levels. Both patients had no other significant discomfort. Literature search retrieved 0 Chinese literature and 22 English literatures. In addition to the 2 cases reported in this study, a total of 51 patients were identified as CMRD caused by SAR1B gene variations. Twenty-one types of SAR1B variants 10 missense, 4 nonsense, 3 frameshift, 1 in-frame deletion, 1 splice, 1 gross deletion, and 1 gross insertion-deletion were found among the 51 CMRD cases. Among all the patients, 49 cases had lipid malabsorption (43 cases had diarrhea or fatty diarrhea, 17 cases had vomiting, and 12 cases had abdominal distension), 45 cases had lipid soluble Vitamin deficiency (43 cases had Vitamin E deficiency, 10 cases had Vitamin A deficiency, 9 case had Vitamin D deficiency, and 5 cases had Vitamin K deficiency), 35 cases had failure to thrive, 32 cases had liver involvement (32 cases had elevated transaminases, 5 cases had fatty liver, and 3 cases had hepatomegaly), 29 cases had white small intestinal mucosa under endoscopy, and 17 cases had elevated creatine kinase, 14 cases had neuropathy, 5 cases had ocular lesions, 2 cases had acanthocytosis, 1 case had decreased cardiac ejection fraction, and 1 case was symptom-free. Conclusions: Early infancy failure to thrive and lipid malabsorption are common issues for CMRD patients. The laboratory tests are characterized by hypocholesterolemia with or without fat-soluble Vitamin deficiency, elevated liver enzymes and (or) creatine kinase. Currently, missense variations are frequent among the primarily homozygous SAR1B genotypes that have been described.

[Molecular diagnostic strategies and management of dust mite allergy].

Dust mites are one of the most important allergens, widely distributed around the world, especially in household environments. Dermatophagoides pteronyssinus, Dermatophagoides farinae and Blomia tropicalis are the most common species of dust mites. There are more than 35 known sensitization components of dust mites, among which Der p 1, Der p 2 and Der p 23 are the major components. Clinically, allergen skin test and serum specific immunoglobulin E (sIgE) detection are widely used in the preliminary diagnosis of dust mite allergy. However, these methods cannot accurately identify specific dust mite sensitization components. Considering that there are significant differences in the allergenic components of dust mites in different regions and populations, component-resolved diagnosis of dust mite is particularly important in accurately determining the allergenic components. This is not only of guiding significance for allergen avoidance, but also important for determining the immunotherapy regimen for dust mites. In order to strengthen the understanding of the molecular diagnosis of dust mites and promote the integration of allergy science in China with the international standards, this article interprets the "Allergy Molecular Allergology User's Guide 2.0" published recently by the European Academy of Allergy and Clinical Immunology.

Effects of dietary niacinamide and CP concentrations on the nitrogen excretion, growth performance, and meat quality of pigs.

Reducing the dietary CP concentration in the formulation of low-protein diets without adverse effects on animal growth performance and meat quality remains challenging. In this study, we investigated the effects of nicotinamide (NAM) on the nitrogen excretion, growth performance, and meat quality of growing-finishing pigs fed low-protein diets. To measure the nitrogen balance, we conducted two trials: in nitrogen balance trial 1, four crossbred (Duroc × Landrace × Large White) barrows (40 ± 0.5 kg BW) were used in a 4 × 4 Latin square design with four diets and periods. The diets consisted of a basal diet + 30 mg/kg NAM (a control dose), basal diet + 90 mg/kg NAM, basal diet + 210 mg/kg NAM, and basal diet + 360 mg/kg NAM. In nitrogen balance trial 2, another four barrows (40 ± 0.5 kg BW) were used in a 4 × 4 Latin square design. The diets consisted of a basal diet + including 30 mg/kg NAM (control), basal diet + 360 mg/kg NAM, low-protein diet + 30 mg/kg NAM, and low-protein diet + 360 mg/kg NAM. To measure growth performance, two trials were conducted. In growth performance trial 1, 40 barrows (37.0 ± 1.0 kg) were randomly allocated to one of four dietary treatments (n = 10 per group), whereas in growth performance trial 2, 300 barrows (41.4 ± 2.0 kg) were randomly allocated to one of four dietary treatments, with each dietary treatment conducted in five repetitions with 15 pigs each. The four diets in the two growth performance trials were similar to those in nitrogen balance trial 2. Supplementing the diet with 210 or 360 mg/kg NAM reduced urinary nitrogen excretion and total nitrogen excretion and increased nitrogen retention comparted with the control diet (P < 0.05). Compared with the control diet, the low-protein diet with 360 mg/kg NAM reduced faecal, urinary, and total nitrogen excretion (P < 0.05) without affecting nitrogen retention and average daily gain (P > 0.05). Pigs fed the low-protein diet with 360 mg/kg NAM showed a decreased intramuscular fat content in the longissimus thoracis muscle when compared with pigs fed the control diet (P > 0.05). Our results suggest NAM as a suitable dietary additive to reduce dietary CP concentration, maximise nitrogen retention and growth performance, and decrease fat deposition in pigs.

[Analysis of specific sIgE detection of house dust mites and aspergillus fumigatus in 2 535 patients with respiratory allergic diseases and respiratory infectious diseases in the Guangzhou area].

Objective: To compare the specific IgE positive rates of the patients between allergic respiratory diseases and respiratory infectious diseases in Guangzhou, the relationship between the co-sensitization of house dust mite (HDM) allergen and Aspergillus fumigatus (AF) allergen and asthma, allergic rhinitis with asthma, pneumonia, upper respiratory infections, bronchitis, serum total immunoglobulin E (total Immunoglobulin E, tIgE) and age were analyzed, to provide the basis for the prevention and treatment of respiratory allergic diseases and respiratory infectious diseases in this area. Methods: A total of 2 535 patients with confirmed respiratory allergic diseases and respiratory infectious diseases were selected retrospectively from the outpatient or inpatient department of the First Affiliated Hospital of Guangzhou Medical University from April 2017 to June 2021 and detected HDM and AF specific IgE (sIgE) by the ImmunoCAP system. The age range was 1 to 89 years. The median age was 5 years. The average age was 9. ≤3 years old group n=894, 4-6 years old group n=721, 7-18 years old group n=615, 19-49 years old group n=207, >49 years old group n=98. There were 1 596 males (62.96%) and 939 females (37.04%). There were 1 279 cases of allergic diseases and 1 256 cases of respiratory infectious diseases. The different disease groups were divided into asthma group (411 cases), allergic rhinitis group (458 cases), allergic rhinitis combined with asthma group (410 cases), pneumonia group (463 cases), upper respiratory tract infection group (299 cases) and bronchitis group (494 cases). The difference of specific IgE (sIgE) and tIgE between HDM and AF was analyzed. For statistical analysis, continuous variables were tested by Mann-Whitney U. Classification data by chi-square test or Fisher's exact test. Results: 1 313 (51.79%) patients were sIgE positive for HDM allergen, 65 (2.56%) were sIgE positive for AF allergen, and 50 (1.97%) were both positive. In the respiratory allergic disease group, 877 cases (68.57%,877/1 279) were positive for HDM allergen sIgE, 57 cases (4.46%,57/1 279) were positive for AF allergen sIgE, and 44 cases (3.44%,44/1 279) were both positive; 436 cases (34.71%,436/1 256) of respiratory infectious diseases were positive for HDM allergen sIgE, 8 cases (0.64%,8/1 256) were positive for AF allergen sIgE, and 6 cases (0.48%,6/1 256) were both positive. In monosensitization, the HDM allergen sIgE sensitization rate was the highest in the allergic rhinitis & asthma group, at 80.24% (329/410). The positive rate of HDM allergen sIgE in male patients was 53.76%(858/1 596), and the positive rate in female patients was 46.22%(434/939), and the difference between the two was statistically significant (χ2=13.449, P<0.001). In polysensitization, asthma patients (5.35%,22/411) had the highest positive rate of HDM sensitization with AF, followed by allergic rhinitis patients (3.06%,14/458), allergic rhinitis with asthma (1.95%,8/410). The positive rate of respiratory infectious diseases such as pneumonia (0.43%,2/463), upper respiratory infections (0.33%,1/299), and bronchitis (0.61%,3/494) with AF was extremely low. The positive rate of HDM combined with AF in infants(≤3 years old group,0.34%, 3/894; 4-6 years old group, 0.97%, 7/721)was significantly lower than that in teenagers and adults(7-18 years old group,3.58%, 22/615; 19-49 years old group,6.28%, 13/207;>49 years old group,5.10%, 5/98).In the patients with HDM and AF combined sensitization, HDM sIgE levels were distributed in all grades, and AF sIgE levels were mainly in grades 1, 2, and 3. Conclusion: The positive rate of HDM combined with AF was higher in patients with respiratory allergic diseases such as asthma, allergic rhinitis, and allergic rhinitis combined with asthma, suggesting that clinical attention should be paid to the combination of HDM and AF in patients with asthma and allergic rhinitis, especially adults, more likely to be combined with AF.

[Research on allergen detection technology and allergic disease diagnosis strategy in China].

In China, the prevalence of allergic diseases is increasing, but allergy research and allergy diagnosis technologies are still in their infancy, posing certain hurdles to clinical diagnosis and treatment. The detection of allergen is essential for the prevention and treatment of allergic diseases. Clarifying the aetiology and risk factors of allergic diseases is critical in order to provide targeted environmental control and therapy for allergic diseases, as well as reduce patient pain and financial burden. In this study, allergen detection technologies and diagnosis strategies are described in order to provide direction for clinicians and laboratory technicians, improve allergic diseases diagnosis, and guide allergic disease therapy.

[Analysis of serum allergen-specific IgE in children with respiratory allergic disease in Guangdong].

Objective: To investigate the common allergens in children with respiratory allergic diseases in Guangdong and provide the basis for preventing and treating respiratory allergic diseases. Methods: A total of 254 serum samples were collected retrospectively from children with allergic rhinitis and (or) asthma in the pediatric clinic of the First Affiliated Hospital of Guangzhou Medical University or Boai Hospital of Zhongshan City from August 2019 to August 2020 and tested positive for at least one allergens by ImmunoCAP 1000 or skin prick test. There were 173 males and 81 females, with a median age (interquartile interval) of 7 (4.00, 9.75) years. Specific IgE against eighteen common allergen were tested by magnetic particle chemiluminescence immunoassay. Statistical analysis using χ2 test. Results: The positive rate of Dermatophagoides pteronyssinus (85.0%, 215/254) was the highest, followed by D. farinae (83.5%, 212/254), egg white (19.3%, 49/254), milk (14.6%, 37/254), German cockroach (14.2%, 36/254) and ox-eye daisy (12.6%, 32/254). Dust mites were the most common allergens in patients with allergic rhinitis / allergic asthma / allergic rhinitis and asthma. The positive rates of D. pteronyssinus (95.4% vs. 91.7% vs. 71.0%, χ2=23.257, P<0.001), D. farinae (95.4% vs. 90.6% vs. 67.7%, χ2=26.916, P<0.001), mugwort (10.8% vs. 9.4% vs. 1.1%, χ2=7.535, P=0.023), and ox-eye daisy (21.5% vs. 14.6% vs. 4.3%, χ2=10.876, P=0.004) in allergic rhinitis combined with asthma group and allergic rhinitis group were significantly higher than those in allergic rhinitis group. The positive rates of D. pteronyssinus (96.2% vs. 84.0% vs. 53.5%, χ2=46.343, P<0.001), D. farinae (96.2% vs. 81.5% vs. 48.8%, χ2=52.756, P<0.001) and German cockroach (17.7% vs. 14.8% vs. 2.3%, χ2=6.313, P=0.043) in > 6 years old group were significantly higher than those in > 3 years old group and > 3-6 years old group, while the sIgE positive rates of egg white (41.9% vs. 21.0% vs. 10.8%, χ2=20.281, P<0.001) and milk (41.9% vs. 16.0% vs. 4.6%, χ2=36.227, P<0.001) in ≤3 years old group were significantly higher than those in > 3-6 years old group and > 6 years old group. In addition, there were significant positive correlations between D. pteronyssinus and D. farinae, mugwort and ox-eye daisy, cat dander and horse dander, ox-eye daisy and dandelion, egg white and milk, timothy grass and sesame (rs≥0.511, P < 0.01). With the increase of the class of house dust mite allergen, the sIgE positive rates of mugwort (0.0% vs. 1.5% vs. 10.8%, χ2=9.714, P=0.008), ox-eye daisy (0.0% vs. 4.4% vs. 19.6%, χ2=16.195, P<0.001), cat dander (0.0% vs. 7.4% vs. 18.2%, χ2=11.459, P=0.003) and horse dander (0.0% vs. 1.5% vs. 15.5%, χ2=15.443, P < 0.001) increased significantly, while the sIgE positive rate of milk (29.0% vs. 16.2% vs. 10.1%, χ2=8.792, P=0.012) decreased significantly. Conclusions: Dust mite is the main allergen in children with respiratory allergic diseases in Guangdong, and ox-eye daisy is the main pollen allergen. The sensitization patterns of allergens varied in different ages and diseases, and children with respiratory allergic diseases should regularly detect allergens and pay attention to avoid them.

Reducing protein content in the diet of growing goats: implications for nitrogen balance, intestinal nutrient digestion and absorption, and rumen microbiota.

Reducing dietary CP content is an effective approach to reduce animal nitrogen excretion and save protein feed resources. However, it is not clear how reducing dietary CP content affects the nutrient digestion and absorption in the gut of ruminants, therefore it is difficult to accurately determine how much reduction in dietary CP content is appropriate. This study was conducted to investigate the effects of reduced dietary CP content on N balance, intestinal nutrient digestion and absorption, and rumen microbiota in growing goats. To determine N balance, 18 growing wether goats (25.0 ± 0.5 kg) were randomly assigned to one of three diets: 13.0% (control), 11.5% and 10.0% CP. Another 18 growing wether goats (25.0 ± 0.5 kg) were surgically fitted with ruminal, proximate duodenal, and terminal ileal fistulae and were randomly assigned to one of the three diets to investigate intestinal amino acid (AA) absorption and rumen microbiota. The results showed that fecal and urinary N excretion of goats fed diets containing 11.5% and 10.0% CP were lower than those of goats fed the control diet (P < 0.05). When compared with goats fed the control diet, N retention was decreased and apparent N digestibility in the entire gastrointestinal tract was increased in goats fed the 10% CP diet (P < 0.05). When compared with goats fed the control diet, the duodenal flow of lysine, tryptophan and phenylalanine was decreased in goats fed the 11.5% CP diet (P < 0.05) and that of lysine, methionine, tryptophan, phenylalanine, leucine, glutamic acid, tyrosine, essential AAs (EAAs) and total AAs (TAAs) was decreased in goats fed the 10.0% CP diet (P < 0.05). When compared with goats fed the control diet, the apparent absorption of TAAs in the small intestine was increased in goats fed the 11.5% CP diet (P < 0.05) and that of isoleucine, serine, cysteine, EAAs, non-essential AAs, and TAAs in the small intestine was increased in goats fed the 10.0% CP diet (P < 0.05). When compared with goats fed the control diet, the relative richness of Bacteroidetes and Fibrobacteres was increased and that of Proteobacteria and Synergistetes was decreased in the rumen of goats fed a diet with 10.0% CP. In conclusion, reducing dietary CP content reduced N excretion and increased nutrient utilization by improving rumen fermentation, enhancing nutrient digestion and absorption, and altering rumen microbiota in growing goats.

Identifying substance misuse in primary care: TAPS Tool compared to the WHO ASSIST.

BACKGROUND: There is a need for screening and brief assessment instruments to identify primary care patients with substance use problems. This study's aim was to examine the performance of a two-step screening and brief assessment instrument, the TAPS Tool, compared to the WHO ASSIST. METHODS: Two thousand adult primary care patients recruited from five primary care clinics in four Eastern US states completed the TAPS Tool followed by the ASSIST. The ability of the TAPS Tool to identify moderate- and high-risk use scores on the ASSIST was examined using sensitivity and specificity analyses. RESULTS: The interviewer and self-administered computer tablet versions of the TAPS Tool generated similar results. The interviewer-administered version (at cut-off of 2), had acceptable sensitivity and specificity for high-risk tobacco (0.90 and 0.77) and alcohol (0.87 and 0.80) use. For illicit drugs, sensitivities were >0.82 and specificities >0.92. The TAPS (at a cut-off of 1) had good sensitivity and specificity for moderate-risk tobacco use (0.83 and 0.97) and alcohol (0.83 and 0.74). Among illicit drugs, sensitivity was acceptable for moderate-risk of marijuana (0.71), while it was low for all other illicit drugs and non-medical use of prescription medications. Specificities were 0.97 or higher for all illicit drugs and prescription medications. CONCLUSIONS: The TAPS Tool identified adult primary care patients with high-risk ASSIST scores for all substances as well moderate-risk users of tobacco, alcohol, and marijuana, although it did not perform well in identifying patients with moderate-risk use of other drugs or non-medical use of prescription medications. The advantages of the TAPS Tool over the ASSIST are its more limited number of items and focus solely on substance use in the past 3months.

Why not nursing? Factors influencing healthcare career choice among Singaporean students.

BACKGROUND: Internationally, and particularly in Singapore, health education institutions are facing challenges in attracting school leavers to enter nursing courses. OBJECTIVES: To identify the factors influencing the career choice of Singaporean healthcare students and determine the deterrents in choosing nursing as a career choice. DESIGN: An exploratory descriptive qualitative study design was used. PARTICIPANTS: Fifty-nine healthcare students from three higher education institutions were recruited. METHOD: Four nursing and four non-nursing focus groups discussion were held. Interview transcripts were analysed using thematic analysis. RESULTS: Six themes emerged as follows: 'personal interest'; 'prior healthcare exposure'; 'job prospects'; 'academic performance'; 'perceived nature of work'; and 'social influences'. DISCUSSION: The personal interests to help and care along with prior healthcare exposures were found to influence the students' choice. Job prospects such as the ease of getting a job, job stability, and job salary were considered. Nursing was perceived as a course for students with poor academic ability. Misconceptions about the nature of work and a lack of social recognition were identified as deterring factors in students' choice of nursing as a career. CONCLUSIONS: An understanding of the career decision process among healthcare students enables educational leaders and policy-makers to enhance the focus of nursing recruitment strategies. IMPLICATIONS FOR NURSING AND/OR HEALTH POLICY: Strategies for nursing recruitment in Singapore must include creating more opportunities for exposure to nursing in early school years, reviewing the admission policies for nursing programmes to attract academically abled students, ensuring that nursing graduates' salaries are comparable with other healthcare graduates, promoting a better understanding on the role of a registered nurse and its career developments, and providing support for those who are interested in nursing but are faced with career decision-making difficulties due to their families.

Why not nursing? A systematic review of factors influencing career choice among healthcare students.

BACKGROUND: A global shortage of healthcare professionals calls for effective recruitment and retention strategies. The nursing profession faces greater staffing shortages compared with other healthcare professions. Identifying these factors for choosing a career in health care is an important step in structuring future nursing recruitment strategies. AIM: This systematic review examined the motivations for choosing a career in health care, then compared them to factors that influence the choice to pursue a career in nursing. METHODS: A literature search of the CINAHL, PubMed, Web of Science and Scopus databases for articles published between 2002 and 2013 was conducted. The search included studies that focused on factors influencing career choice among undergraduate medicine, dentistry, pharmacy and nursing students. RESULTS: A total of 29 papers were included in the review. The themes and subthemes that emerged from this review included: (1) intrinsic factors, including a desire to help others and a personal interest in health care, (2) extrinsic factors, such as financial remuneration, job security, professional prestige and job autonomy, (3) socio-demographic factors such as gender and socio-economic status, and (4) interpersonal factors, encompassing the influence of family and other professional individuals. DISCUSSION: Healthcare professionals were generally motivated by intrinsic factors. However, public perceptions of nursing as a low-paying and low-status job have significantly hindered the participants' choice to pursue it as a career. CONCLUSION AND IMPLICATIONS FOR NURSING AND/OR HEALTH POLICY: Nursing institutions could provide more platforms to help school leavers better understand the nursing career. In turn, hospital administrators could invite parents to nursing career fairs, increase financial remuneration for nurses, and provide decision-making avenues aimed at recruiting and retaining more nurses.

RpA, an extracellular protease similar to the metalloprotease of serralysin family, is required for pathogenicity of Ralstonia pickettii.

AIM: To investigate the biochemical and functional properties of an extracellular protease, RpA, in Ralstonia pickettii WP1 isolated from water supply systems. METHODS AND RESULTS: An extracellular protease was identified and characterized from R. pickettii WP1. A mutant strain WP1M2 was created from strain WP1 by mini-Tn5 transposition. The culture filtrates from WP1M2 had a lower cytotoxic effect than the parental WP1 on several mammalian cell lines. Cloning and sequence analysis revealed the Tn5 transposon inserted at a protease gene (rpA) which is 81% homologous to prtA and aprX genes of Pseudomonas fluorescens. The rpA gene encodes a 482-residue protein showing sequence similarity to metalloproteases of the serralysin family. The RpA protein was expressed in Escherichia coli using a pET expression vector and purified as a 55 kDa molecular weight protein. Furthermore, the protease activity of RpA was inhibited by protease inhibitor and heat treatment. CONCLUSIONS: The in vitro cytotoxic activity of R. pickettii culture filtrates was attributed to RpA protease. SIGNIFICANCE AND IMPACT OF THE STUDY: An extracellular protease, RpA, was identified from R. pickettii WP1 isolated from water supply system. The RpA metalloproteases is required for the pathogenicity of R. pickettii to mammalian cell lines.

Substance use disorders and co-morbidities among Asian Americans and Native Hawaiians/Pacific Islanders.

BACKGROUND: Asian Americans (AAs) and Native Hawaiians/Pacific Islanders (NHs/PIs) are the fastest growing segments of the US population. However, their population sizes are small, and thus AAs and NHs/PIs are often aggregated into a single racial/ethnic group or omitted from research and health statistics. The groups' substance use disorders (SUDs) and treatment needs have been under-recognized. METHOD: We examined recent epidemiological data on the extent of alcohol and drug use disorders and the use of treatment services by AAs and NHs/PIs. RESULTS: NHs/PIs on average were less educated and had lower levels of household income than AAs. Considered as a single group, AAs and NHs/PIs showed a low prevalence of substance use and disorders. Analyses of survey data that compared AAs and NHs/PIs revealed higher prevalences of substance use (alcohol, drugs), depression and delinquency among NHs than among AAs. Among treatment-seeking patients in mental healthcare settings, NHs/PIs had higher prevalences of DSM-IV diagnoses than AAs (alcohol/drug, mood, adjustment, childhood-onset disruptive or impulse-control disorders), although co-morbidity was common in both groups. AAs and NHs/PIs with an SUD were unlikely to use treatment, especially treatment for alcohol problems, and treatment use tended to be related to involvement with the criminal justice system. CONCLUSIONS: Although available data are limited by small sample sizes of AAs and NHs/PIs, they demonstrate the need to separate AAs and NHs/PIs in health statistics and increase research into substance use and treatment needs for these fast-growing but understudied population groups.

Abuse and dependence on prescription opioids in adults: a mixture categorical and dimensional approach to diagnostic classification.

BACKGROUND: For the emerging DSM-V, it has been recommended that dimensional and categorical methods be used simultaneously in diagnostic classification; however, little is known about this combined approach for abuse and dependence. METHOD: Using data (n=37 708) from the 2007 National Survey on Drug Use and Health (NSDUH), DSM-IV criteria for prescription opioid abuse and dependence among non-prescribed opioid users (n=3037) were examined using factor analysis (FA), latent class analysis (LCA, categorical), item response theory (IRT, dimensional), and factor mixture (hybrid) approaches. RESULTS: A two-class factor mixture model (FMM) combining features of categorical latent classes and dimensional IRT estimates empirically fitted more parsimoniously to abuse and dependence criteria data than models from FA, LCA and IRT procedures respectively. This mixture model included a severely affected group (7%) with a comparatively moderate to high probability (0.32-0.88) of endorsing all abuse and dependence criteria items, and a less severely affected group (93%) with a low probability (0.003-0.16) of endorsing all criteria. The two empirically defined groups differed significantly in the pattern of non-prescribed opioid use, co-morbid major depression, and substance abuse treatment use. CONCLUSIONS: A factor mixture model integrating categorical and dimensional features of classification fits better to DSM-IV criteria for prescription opioid abuse and dependence in adults than a categorical or dimensional approach. Research is needed to examine the utility of this mixture classification for substance use disorders and treatment response.

Association of MDMA/ecstasy and other substance use with self-reported sexually transmitted diseases among college-aged adults: a national study.

OBJECTIVES: MDMA/ecstasy use among college students has increased and reportedly leads to risky sexual behaviours. However, little is known about its association with sexually transmitted diseases (STDs). To evaluate this public health concern, this study examined the association between substance use (particularly MDMA) and self-reported STDs (chlamydia, gonorrhoea, herpes and syphilis) among college students and non-students aged 18-22 years (n=20,858). STUDY DESIGN: A cross-sectional data analysis of a national survey. METHODS: Data were drawn from the 2005-2006 National Surveys on Drug Use and Health; a nationally representative survey of non-institutionalized Americans. Self-reported STDs and substance use were assessed by the audio computer-assisted self-interviewing method. The association between MDMA use and STDs was determined while taking into account young adults' use of other substances, healthcare utilization and sociodemographic characteristics. RESULTS: Overall, 2.1% of college students and 2.5% of non-students reported contracting an STD in the past year. MDMA use in the past year was not associated with STDs. Among non-students, onset of MDMA use before 18 years of age increased the odds of past-year STDs. In both groups, alcohol use, marijuana use, female gender and African American race increased the odds of both past-year and lifetime STDs. Additional analyses indicated that, regardless of college-attending status, greater odds of past-year STDs were noted among users of alcohol and drugs, and users of alcohol alone, but not among users of drugs alone. CONCLUSIONS: Alcohol use is a robust correlate of STDs. Irrespective of college-attending status, young women and African Americans have a higher rate of STDs than young men and Whites.

Genetic association of blaSHV-5 with transposable elements IS26 and IS5 in Klebsiella pneumoniae from Taiwan.

A cloned 5,248-bp EcoRI fragment from the Klebsiella pneumoniae transferable plasmid pKP53 (> 70 kb) containing bla(SHV-5) was sequenced. Insertion sequences IS26 and IS5 were found downstream from bla(SHV-5). The DNA sequences of the genetic environment surrounding bla(SHV-5) were homologous to plasmid p1658/97 from Escherichia coli, containing a truncated recF gene and a truncated deoR gene upstream and downstream from bla(SHV-5), respectively. RecF may be involved in bla(SHV-5) translocation to the plasmid by RecF-dependent recombination. This novel genetic environment may be associated with the successful proliferation and/or expression of SHV-5 in K. pneumoniae strains from Taiwan.

Characterisation and molecular epidemiology of extended-spectrum beta-lactamase-producing Enterobacter cloacae isolated from a district teaching hospital in Taiwan.

Enterobacter cloacae (n = 110) isolates from a district hospital in Taiwan were screened for extended-spectrum beta-lactamases (ESBLs). In total, 17 ESBL-producers were identified, based on the combination-disk synergy test using cefotaxime and ceftazidime +/- clavulanic acid. Investigation of ESBL genes in 33 ceftazidime-resistant isolates revealed the SHV-12 gene in the same 17 ESBL-producers. In addition, one isolate also carried the CTX-M-3 gene, and two isolates also carried the CTX-M-9 gene. No major epidemic clone of ESBL-producers was identified by pulsed-field gel electrophoresis. Routine screening for the ESBL phenotype, focusing on ceftazidime-resistant E. cloacae, should be undertaken in this area.

Identification of a novel cephalosporinase (DHA-3) in Klebsiella pneumoniae isolated in Taiwan.

A strain of Klebsiella pneumoniae resistant to cefoxitin and oxyimino-cephalosporins, but susceptible to cefepime, was isolated from an adult patient hospitalised in Taichung, Taiwan. Isoelectric focusing revealed three beta-lactamases with isoelectric points of 5.4, 8.2 and 7.9, respectively. Following PCR with plasmid DNA templates and gene sequencing, these enzymes were shown to correspond to TEM-1, SHV-5 and a novel DHA-1-like enzyme (designated DHA-3). The bla genes for TEM-1 and SHV-5 were transferable, but the bla(DHA-3) gene was non-self-transferable in conjugation experiments. All three bla genes were successfully introduced by electrotransformation into an Escherichia coli recipient (DH5alpha), resulting in a similar resistance profile to that observed in the original donor strain. Other K. pneumoniae strains producing DHA-1-like enzymes have been identified previously in Taiwan, and this report suggests that DHA-type beta-lactamases are continuing to emerge in this country.

Effects of tannic acid and its related compounds on food mutagens or hydrogen peroxide-induced DNA strands breaks in human lymphocytes.

The effect of tannic acid (TA), gallic acid (GA), propyl gallate (PA) and ellagic acid (EA) on DNA damage in human lymphocytes induced by food mutagens [3-amino-1-methyl-5H-pyrido (4,3-b) indole (Trp-P-2) and 2-amino-1-methyl-6-phenylimadazo (4,5-b) pyridine (PhIP) or H2O2 was evaluated by using single-cell electrophoresis (comet assay). The toxicity of these tested compounds (0.1-100 microg/ml) on lymphocytes was not found. These compounds did not cause DNA strand breaks at lower concentrations of 0.1-10 microg/ml. At a concentration of 100 microg/ml, TA and GA exhibited slight DNA damage, whereas PA and EA showed no DNA strand breaks. TA and its related compounds decreased the DNA strand breaks induced by Trp-P-2, PhIP or H2O2 at concentrations of 0.1-10 microg/ml. DNA repair enzymes endonuclease III (Endo III) and formamidopyrimidine-DNA glycoslase (FPG)] were used to examine the levels of oxidised pyrimidines and purines in human lymphocytes induced by H2O2. All the compounds at 10 microg/ml can reduce the level of FPG sensitive sites. However, only EA inhibited the formation of EndoIII sensitive sites. The results indicated that these compounds can enhance lymphocytes resistance towards DNA strand breaks induced by food mutagens or H2O2 in vitro.

Production of a recombinant human basic fibroblast growth factor with a collagen binding domain.

Basic fibroblast growth factor (bFGF) is a potent in vitro mitogen for capillary endothelial cells, stimulates angiogenesis in vivo, and may participate in tissue repair. Basic FGF is found in abundance in tissues such as brain, kidney, and cartilage. This study reports the expression, purification, and renaturation of a biologically active human basic fibroblast growth factor fusion protein (hbFGF-F1) from Escherichia coli. A prokaryotic expression vector was engineered to produce a tripartite fusion protein consisting of a purification tag, a protease-sensitive linker and collagen binding domain, and a cDNA sequence encoding the active fragment of hbFGF. The expressed hbFGF-F1 and hbFGF-F2 (it contains the collagen binding domain), located in inclusion bodies, were solubilized with 6 M guanidine-HCl and renatured by a glutathione redox system and protracted dialysis under various experimental conditions. The purification of the recombinant proteins was achieved by binding the His-tag of the fusion protein on a nickel-nitrilotriacetic acid metal chelate column. The biological activity of the recombinant growth factor was demonstrated by its ability to stimulate proliferation of human vein endothelial cells, monitored by [3H]thymidine incorporation, where commercial recombinant human bFGF (rhbFGF) served as a positive control. Purified rhbFGF-F1 and rhbFGF-F2 constructs exhibited proliferative activity comparable to commercial rhbFGF. The high-affinity binding was demonstrated by the binding of [3H]collagen to the rhbFGF-F2 protein immobilized on a Ni-nitrilotriacetic acid column. The rhbFGF-F2 fusion protein bound to collagen-coated surfaces with high affinity. Taken together, these results demonstrate that biologically active rhbFGF fusion proteins can be recovered from transformed bacteria by oxidative refolding; thus, providing a means for their high-yield production, purification, and renaturation from microorganisms. Furthermore, we demonstrate that the auxiliary collagen binding domain effectively targets the recombinant growth factor to type I collagen. These studies advance the technology necessary to generate large quantities of targeted bFGF fusion proteins for specific biomedical applications.

Engineering, expression, and renaturation of a collagen-targeted human bFGF fusion protein.

Basic fibroblast growth factor (bFGF) is a potent in vitro mitogen for capillary endothelial cells, stimulates angiogenesis in vivo, and may participate in tissue repair. Basic FGF is found in abundance in tissues such as brain, kidney and cartilage. This study reports the expression, purification, and renaturation of a biologically active human basic fibroblast growth factor fusion protein (hbFGF-F1) from Escherichia coli. A prokaryotic expression vector was engineered to produce a tripartite fusion protein consisting of (i) a purification tag, (ii) a protease-sensitive linker/collagen-binding domain, and (iii) cDNA sequence encoding the active fragment of hbFGF. The expressed hbFGF-F1 and hbFGF-F2 (it contains a collagen-binding domain), located in inclusion bodies, were solubilized with 6 M guanidine-HCl and renatured using a glutathione redox system and protracted dialysis under various experimental conditions. The purification of the recombinant proteins was achieved by binding the His-tag of the fusion protein on a Ni-NTA metal chelate column. The biological activity of the recombinant growth factors was demonstrated by their ability to stimulate proliferation of human vein endothelial cells (HVEC), monitored by [3H]-thymidine incorporation, where commercial recombinant human bFGF (rhbFGF) served as a positive control. Purified rhbFGF-F1 and rhbFGF-F2 constructs exhibited proliferative activity comparable to commercial rhbFGF. Binding of the renatured hbFGF-F2 fusion protein to collagen was demonstrated by stable binding on a collagen-conjugated Sephadex-G15 column. The high affinity binding was also demonstrated by the binding of [3H]-collagen to the rhbFGF-F2 protein immobilized on a Ni-NTA column. The rhbFGF-F2 fusion protein bound to collagen coated surfaces with high affinity but exhibited comparatively lower biological activity than the fusion protein in solution, suggesting a potentially latent configuration. Taken together, these results demonstrate that biologically active rhbFGF fusion proteins can be recovered from transformed bacteria by oxidative refolding; thus, providing a means for its high-yield production, purification, and renaturation from microorganisms. Furthermore, we demonstrate that the auxiliary collagen-binding domain effectively targets the recombinant growth factor to type I collagen. The clinical effect of rhbFGF-F2 on wound healing is also studied in streptozotocin-induced diabetic rats and evaluated by histological examination comparing with rhbFGF-F1 and commercial bFGF effects. The highly beneficial effects of rhbFGF-F2 on wound healing is suggested to be due to its extremely potent angiogenesis and granulation tissue formation activities, leading to a rapid reepithelialization of the wound. Topical application of rhbFGF-F2 mixed with type I collagen is a more effective method in accelerating closure of full-thickness excisional skin-wound in diabetic rats when compared with the fusion protein alone or commercial hbFGF at the same doses. These studies advance the technology necessary to generate large quantities of targeted bFGF fusion proteins as well as to develop new strategies for specific biomedical applications.