Correction: Alkaline Ceramidase 3 Deficiency Results in Purkinje Cell Degeneration and Cerebellar Ataxia Due to Dyshomeostasis of Sphingolipids in the Brain.

[This corrects the article DOI: 10.1371/journal.pgen.1005591.].

Expression profile of long noncoding RNAs in human papillary thyroid carcinoma.

Long non-coding RNAs (lncRNAs) are strongly associated with cancer biology. The objective of this study is to investigate the expression profile of lncRNAs in human papillary thyroid carcinoma (PTC), and to understand the biological role of lncRNAs and their involvement in PTC oncogenesis. The lncRNAs and messenger RNAs (mRNAs) expression in human PTC and paired adjacent non-cancerous thyroid (NCT) tissues were studied by micro-array, and quantitative real-time polymerase chain reaction (qRT-PCR) validated five differentially expressed lncRNAs. We identified 2,925 significantly differentially expressed lncRNAs with potential roles in PTC, (absolute fold change >2.0; p<0.05). Of these, 1,922 were up-regulated and 933 down-regulated and the qRT-PCR results agreed with micro-array results. Gene ontology (GO) enrichment and pathway analysis then investigated gene function and identified significantly enriched pathways in differentially expressed mRNA's. Many of these pathways were related to cancer, including 60 genes associated with "pathways in cancer" and 34 linked to "proteoglycans in cancer". Co-expression network and target prediction analysis of lncRNAs revealed that TCONS_00020457 can have important roles in PTC. In conclusion, the results of this study indicate that lncRNAs can be important regulators in PTC tumorigenesis and provide understanding of the function and mechanism of lncRNAs related to human papillary thyroid carcinoma.

Alkaline Ceramidase 3 Deficiency Results in Purkinje Cell Degeneration and Cerebellar Ataxia Due to Dyshomeostasis of Sphingolipids in the Brain.

Dyshomeostasis of both ceramides and sphingosine-1-phosphate (S1P) in the brain has been implicated in aging-associated neurodegenerative disorders in humans. However, mechanisms that maintain the homeostasis of these bioactive sphingolipids in the brain remain unclear. Mouse alkaline ceramidase 3 (Acer3), which preferentially catalyzes the hydrolysis of C18:1-ceramide, a major unsaturated long-chain ceramide species in the brain, is upregulated with age in the mouse brain. Acer3 knockout causes an age-dependent accumulation of various ceramides and C18:1-monohexosylceramide and abolishes the age-related increase in the levels of sphingosine and S1P in the brain; thereby resulting in Purkinje cell degeneration in the cerebellum and deficits in motor coordination and balance. Our results indicate that Acer3 plays critically protective roles in controlling the homeostasis of various sphingolipids, including ceramides, sphingosine, S1P, and certain complex sphingolipids in the brain and protects Purkinje cells from premature degeneration.

Coronary microcirculatory dysfunction is associated with left ventricular dysfunction during follow-up after STEMI.

BACKGROUND: Coronary microvascular resistance is increased after primary percutaneous coronary intervention (PCI) for ST-elevation myocardial infarction (STEMI), which may be related in part to changed left ventricular (LV) dynamics. Therefore we studied the coronary microcirculation in relation to systolic and diastolic LV function after STEMI. METHODS: The study cohort consisted of 12 consecutive patients, all treated with primary PCI for a first anterior wall STEMI. At 4 months, we assessed pressure-volume loops. Subsequently, we measured intracoronary pressure and flow velocity and calculated coronary microvascular resistance. Infarct size and LV mass were assessed using magnetic resonance imaging. RESULTS: Patients with an impaired systolic LV function due to a larger myocardial infarction showed a higher baseline average peak flow velocity (APV) than the other patients (26 ± 7 versus 17 ± 5 cm/s, p = 0.003, respectively), and showed an impaired variable microvascular resistance index (2.1 ± 1.0 versus 4.1 ± 1.3 mmHg cm(-1)∙s(-1), p = 0.003, respectively). Impaired diastolic relaxation time was inversely correlated with hyperaemic APV (r = -0.56, p = 0.003) and positively correlated with hyperaemic microvascular resistance (r = 0.48, p = 0.01). LV dilatation was associated with a reduced variable microvascular resistance index (r = 0.78, p = 0.006). CONCLUSION: A larger anterior myocardial infarction results in impaired LV performance associated with reduced coronary microvascular resistance variability, in particular due to higher coronary blood flow at baseline in these compromised left ventricles.

[Association between socioeconomic status and overweight/obesity in Yi people, Sichuan province].

Objective: To explore the association between socioeconomic status (SES) and overweight/obesity in Yi people in Sichuan province. Methods: A cross-sectional study was conducted in Liangshan Yi Autonomous Prefecture in 2015. Stratified cluster sampling method was used to enroll Yi farmers and rural-to-urban Yi migrants aged 20-80 years. SES was measured by education level, personal annual income, and compound SES index. Unconditional logistic regression models were used to determine the association between SES and overweight/obesity (BMI≥24.0 kg/m(2)). Results: 1 894 Yi farmers and 1 162 rural-to urban migrants were included in the analysis. After adjustment for age, smoking, drinking and physical activity, compared with illiteracy, OR for farmer males with higher education level (primary or junior school, senior high school or higher) were 1.71 (95%CI: 1.13-2.58) and 4.15 (95%CI: 2.10-8.22). Compared with lower income group (<5 000 CNY/y), the higher income group had increased risk (OR=1.66, 95%CI: 1.12-2.44). For farmer males with medium and high SES level, the risk of overweight/obesity were 1.65 (95%CI: 1.02-2.67) and 3.26 (95%CI: 1.97-5.42) compared with low level of SES. For farmer females, the risk increased with the higher income, with OR as 1.49 (95%CI: 1.10-2.02). Compared with low SES level, farmer females with medium level of SES was associated with 1.47 (95%CI: 1.11-1.95) times higher risk of overweight/obesity. In Yi migrants, the association between SES and overweight/obesity was not found. Conclusion: Socioeconomic status was positively associated with overweight/obesity only in Yi farmers.

Lack of evidence of mastitis as a causal factor for postpartum dysgalactia syndrome in sows.

To investigate the prevalence of mastitis in sows suffering from postpartum dysgalactia syndrome (PDS), we examined milk constituents and concentrations of lipopolysaccharides (LPS) obtained from the udder vein (v. epigastrica). As part of a case-cohort study, 109 sows were monitored daily from 60 h antepartum (a.p.) to 36 h postpartum (p.p.). Over time, 38 sows were diagnosed with PDS (PDS+) and were retrospectively matched with 38 healthy sows (PDS-). The study period was divided into 7 smaller time periods (A, B, C, D, E, F, G, H, and E), allowing the studied values, in period B to G, to be compared with period A that served as a baseline, and PDS+ and PDS- sows were compared within the time periods. All sows were subjected to a thorough daily clinical examination and blood was sampled from v. epigastrica for LPS detection. Milk samples were obtained for bacteriological evaluation and detection of N-acetyl-beta-d-glucosaminidase (NAGase), lactate dehydrogenase (LDH), ß-glucuronidase (ß-glu; for evaluation of mastitis), isocitrate (isoC), free glucose, uric acid (UA; for evaluation of the mammary energy status), ß-hydroxybutyrate acid (BHBA; for evaluation of ketosis), and milk urea (for evaluation of the protein status). The results revealed that PDS+ sows had decreased concentrations of UA in milk (P < 0.0001), increased heart rates (P < 0.01), increased mammary edema (P < 0.05), and prolonged capillary refill time in the vulvar mucosa (P < 0.01) compared with PDS- sows. Compared with baseline, feces became more solid 0 to 36 h p.p. (P < 0.0001) and the respiration rate decreased 12 to 24 h p.p. (P < 0.0001) for both PDS+ and PDS- sows. No differences were found between PDS+ and PDS- sows for severe bacterial infections, concentrations of LPS in blood or LDH, NAGase, BHBA, free glucose, isoC, or urea in milk. Concentrations of LPS in blood were not associated with signs of mastitis or edema in the mammary glands. However, a difference over time was seen for redness (P < 0.0001), warmth (P < 0.0001), and hardness (P < 0.05) of the 6 most anterior glands in both PDS+ and PDS- sows from 60 h a.p to 36 h p.p. The PDS- sows had greater concentrations of ß-glu than the PDS+ sows, but no change over time was demonstrated for this marker. In conclusion, signs of mastitis were not consistently linked to PDS in sows. However, the cardiovascular system seemed to be compromised in PDS+ sows and the cause should be investigated to elucidate the pathogenesis of PDS.

Feasibility study of an online modifiable Enhanced Recovery After Surgery protocol with specific focus on opioid avoidance.

BACKGROUND: The high and increasing rate of opioid use is a serious issue in the Western world affecting the population's physical and mental health. In most cases, opioid dependency starts with prescriptions by medical professionals, so efforts aimed at reducing in-hospital opioid use should result in less long-term dependency. The aim of the current study was to evaluate the feasibility of implementing an opioid-scarce protocol as part of a new online modifiable Enhanced Recovery After Surgery (mERAS) programme. METHODS: A single-centre retrospective study was conducted comparing a cohort treated under the new opioid-scarce mERAS protocol (n = 96; May 2018-Nov 2018) to those treated under the original ERAS protocol (n = 84; November 2017-April 2018). The primary outcome was the quantity and duration of opioid use. RESULTS: Fewer patients used fentanyl via intravenous patient-controlled analgesia in the mERAS group (54% versus 70%; P = 0.03). The mERAS group was also less likely to use oral oxycodone (80% versus 99%; P < 0.0001) and for a shorter duration (median 3 versus 5 days; P = 0.0002). More local anaesthetic transversus abdominis plane catheters were used in the mERAS group (34% versus 6% in the control group; P < 0.0001). CONCLUSION: Opioid use can be significantly reduced after elective colorectal surgery by employing an opioid-scarce ERAS protocol. Further data is required to confirm the clinical benefits of this approach.

Effect of feeding allowance level on embryonic survival, IGF-1, insulin, GH, leptin and progesterone secretion in early pregnancy gilts.

The present experiment was conducted to evaluate the effects of feeding allowance level on embryonic survival, uterine development and reproductive hormone secretion in early gestation gilts. A total of 54 F1 crosses of Landrace x Large white gilts were randomly allocated to three treatment groups of high (H, 2 x maintenance), medium (M, 1.2 x maintenance) and low (L, 0.6 x maintenance) feeding level after mating. Blood samples and uterine flushings were collected on day 12, 25 and 35 of pregnancy, and embryonic survival rate was estimated. Concentrations of insulin-like growth factor I (IGF)-1, insulin, growth harmone (GH), leptin and progesterone in serum and uterine flushings were determined by enzyme-linked immunosorbent assay. Embryonic survival was affected by dietary treatment; total number of viable embryos and embryo survival of group M were higher than other groups in the early pregnancy (p < 0.05). Greater foetal weight in M gilts was achieved when gestation advanced to day 35 (p < 0.05), though there was no difference on day 25 of pregnancy among treatments. No appreciable differences were observed in the crown-rump length on day 25 and 35 of pregnancy among groups. Greater weight of uterus and products of conception were identified in M gilts compared with group H and L (p = 0.024 and p = 0.005, respectively) on day 25 of pregnancy. The hormone level was greatly affected by feeding allowance level. In serum, concentrations of IGF-1, leptin and insulin tended to be greater in H than in M and L during the early gestation, while concentrations of GH were greater in M and progesterone were the lowest in H. At the same time, feed allowance level affected the concentration of IGF-1, insulin, GH, leptin and progesterone in uterine flushings. These data demonstrated that feed allowance level after mating has important consequence on embryo survival, embryo development and uterine development, possibly mediated by nutrition level inducing changes in concentrations of reproductive hormones and/or intermediary metabolites in early pregnancy.

Characteristics and short-term outcomes of young women with acute myocardial infarction in Malaysia: a retrospective analysis from the Malaysian National Cardiovascular Database registry.

OBJECTIVE: Young women form a minority but an important group of patients with acute myocardial infarction (MI) as it can potentially cause devastating physical and socioeconomic impact. This study was aimed to investigate the characteristics and outcomes of young women with MI in Malaysia. DESIGN: This is a retrospective analysis of women with ST-elevation MI (STEMI) and non-STEMI (NSTEMI) from 18 hospitals across Malaysia using the Malaysian National Cardiovascular Database registry-acute coronary syndrome (NCVD-ACS). PARTICIPANTS: Women patients diagnosed with acute MI from year 2006 to 2013 were identified and divided into young (age ≤ 45, n=292) and older women (age >45, n=5580). PRIMARY OUTCOME MEASURE: Comparison of demographics, clinical characteristics and in-hospital management was performed between young and older women. In-hospital and 30-day all-cause mortality were examined. RESULTS: Young women (mean age 39±4.68) made up 5% of women with MI and were predominantly of Malay ethnicities (53.8%). They have a higher tendency to present as STEMI compared with older women. Young women have significantly higher rates of family history of premature coronary artery disease (CAD) (20.5% vs 7.8% p<0.0001). The prevalence of risk factors, such as hypertension, diabetes and dyslipidaemia was high in both groups. The primary reperfusion strategy was thrombolysis with no significant differences observed in the choice of intervention for both groups. Other than aspirin, rates of prescriptions for evidence-based medications were similar with >80% prescribed statins and aspirin. The all-cause mortality rates of young women were lower for both in-hospital and 30 days, especially in those with STEMI with adjusted mortality ratio to the older group, was 1:9.84. CONCLUSION: Young women with MI were over-represented by Malays and those with a family history of premature CAD. Preventive measures are needed to reduce cardiovascular risks in young women. Although in-hospital management was similar, short-term mortality outcomes favoured young compared with older women.

In-hospital mortality of cardiogenic shock complicating ST-elevation myocardial infarction in Malaysia: a retrospective analysis of the Malaysian National Cardiovascular Database (NCVD) registry.

OBJECTIVES: Cardiogenic shock (CS) complicating ST-elevation myocardial infarction (STEMI) carries an extremely high mortality. The clinical pattern of this life threatening complication has never been described in Malaysian setting. This study is to investigate the incidence, clinical characteristics and outcome of STEMI patients with CS in our population. DESIGN: A retrospective analysis of STEMI patients from 18 hospitals across Malaysia contributing to the Malaysian National Cardiovascular Database-acute coronary syndrome) registry (NCVD-ACS) year 2006-2013. PARTICIPANTS: 16 517 patients diagnosed of STEMI from 18 hospitals in Malaysia from the year 2006 to 2013. PRIMARY OUTCOME MEASURES: In-hospital and 30 day post-discharge mortality. RESULTS: CS complicates 10.6% of all STEMIs in this study. They had unfavourable premorbid conditions and poor outcomes. The in-hospital mortality rate was 34.1% which translates into a 7.14 times mortality risk increment compared with STEMI without CS. Intravenous thrombolysis remained as the main urgent reperfusion modality. Percutaneous coronary interventions (PCI) in CS conferred a 40% risk reduction over non-invasive therapy but were only done in 33.6% of cases. Age over 65, diabetes mellitus, hypertension, chronic lung and kidney disease conferred higher risk of mortality. CONCLUSION: Mortality rates of CS complicating STEMI in Malaysia are high. In-hospital PCI confers a 40% mortality risk reduction but the rate of PCI among our patients with CS complicating STEMI is still low. Efforts are being made to increase access to invasive therapy for these patients.

Gender differences in mortality among ST elevation myocardial infarction patients in Malaysia from 2006 to 2013.

BACKGROUND: Coronary artery disease (CAD) is one of the leading causes of death in Malaysia. However, the prevalence of CAD in males is higher than in females and mortality rates are also different between the two genders. This suggest that risk factors associated with mortality between males and females are different, so we compared the clinical characteristics and outcome between male and female STEMI patients. OBJECTIVES: To identify the risk factors associated with mortality for each gender and compare differences, if any, among ST-elevation myocardial infarction (STEMI) patients. DESIGN: Retrospective analysis. SETTINGS: Hospitals across Malaysia. PATIENTS AND METHODS: We analyzed data on all STEMI patients in the National Cardiovascular Database-Acute coronary syndrome (NCVD-ACS) registry for the years 2006 to 2013 (8 years). We collected demographic and risk factor data (diabetes mellitus, hypertension, smoking status, dyslipidaemia and family history of CAD). Significant variables from the univariate analysis were further analysed by a multivariate logistic analysis to identify risk factors and compare by gender. MAIN OUTCOME MEASURES: Differential risk factors for each gender. RESULTS: For the 19484 patients included in the analysis, the mortality rate over the 8 years was significantly higher in females (15.4%) than males (7.5%) (P < .001). The univariate analysis showed that the majority of male patients < 65 years while females were >=65 years. The most prevalent risk factors for male patients were smoking (79.3%), followed by hypertension (54.9%) and diabetes mellitus (40.4%), while the most prevalent risk factors for female patients were hypertension (76.8%), followed by diabetes mellitus (60%) and dyslipidaemia (38.1%). The final model for male STEMI patients had seven significant variables: Killip class, age group, hypertension, renal disease, percutaneous coronary intervention and family history of CVD. For female STEMI patients, the significant variables were renal disease, smoking status, Killip class and age group. CONCLUSION: Gender differences existed in the baseline characteristics, associated risk factors, clinical presentation and outcomes among STEMI patients. For STEMI females, the rate of mortality was twice that of males. Once they reach menopausal age, when there is less protection from the estrogen hormone and there are other risk factors, menopausal females are at increased risk for STEMI. LIMITATION: Retrospective registry data with inter-hospital variation.

[Blomia tropicalis is found in a flour warehouse of a food factory in a civil aviation airport].

Objective To investigate whether there is Blomia tropicalis breeding in the flour warehouse of food factory in a civil aviation airport, and, if there is, to observe the morphological structure of the mite. Methods The flour samples were collected from the flour warehouse of food factory in a civil aviation airport. The breeding mites were isolated from the samples by direct microscopy and made into specimens to observe with a light microscope. Results Female Blomia tropicalis was found in the collected flour samples. Under the microscope, the mite was nearly spherical, and it had no tergum and claw. Its apodemeⅠ was developed and connected in the midline. The reproductive hole was located between the foot Ⅲ and IV level and was covered with a pair of oblique genital folds. The foot Ⅲ and Ⅳ had no solenoids. The copulatory pouch was a long, slightly curved tube, extending from the end of the mite. ConclusionsBlomia tropicalis has been found in the flour warehouse of a food factory in the civil aviation airport. The main distinguishing feature of this mite is that it has no tergum, its feet have no pectinate inferior tarsus scales or claw, the feetⅠgenu has only one solenidia, and feet Ⅲ and Ⅳ have no solenidias.

Down-regulation of heme oxygenase-2 is associated with the increased expression of heme oxygenase-1 in human cell lines.

Intracellular heme concentrations are maintained in part by heme degradation, which is catalyzed by heme oxygenase. Heme oxygenase consists of two structurally related isozymes, HO-1 and HO-2. Recent studies have identified HO-2 as a potential oxygen sensor. To gain further insights into the regulatory role of HO-2 in heme homeostasis, we analyzed the expression profiles of HO-2 and the biochemical consequences of HO-2 knockdown with specific short interfering RNA (siRNA) in human cells. Both HO-2 mRNA and protein are expressed in the eight human cancer cell lines examined, and HO-1 expression is detectable in five of the cell lines, including HeLa cervical cancer and HepG2 hepatoma. Down-regulation of HO-2 expression with siRNA against HO-2 (siHO-2) caused induction of HO-1 expression at both mRNA and protein levels in HeLa and HepG2 cells. In contrast, knockdown of HO-1 expression did not noticeably influence HO-2 expression. HO-2 knockdown prolonged the half-life of HO-1 mRNA twofold in HeLa cells. Transient transfection assays in HeLa cells revealed that the 4.5-kb human HO-1 gene promoter was activated with selective knockdown of HO-2 in a sequence-dependent manner. Moreover, HO-2 knockdown caused heme accumulation in HeLa and HepG2 cells only when exposed to exogenous hemin. HO-2 knockdown may mimic a certain physiological change that is important in the maintenance of cellular heme homeostasis. These results suggest that HO-2 may down-regulate the expression of HO-1, thereby directing the co-ordinated expression of HO-1 and HO-2.

Trends in evidence-based treatment and mortality for ST elevation myocardial infarction in Malaysia from 2006 to 2013: time for real change.

BACKGROUND: The administration of evidence-based pharmacotherapy and timely primary percutaneous coronary intervention have been shown to improve outcome in ST elevation myocardial infarction (STEMI). However, implementation remains a challenge due to the limitations in facilities, expertise and funding. OBJECTIVES: To investigate adherence to guideline-based management and mortality of STEMI patients in Malaysia. DESIGN: Retrospective analysis. SETTINGS: STEMI patients from 18 participating hospital across Malaysia included in the National Cardiovascular Database-Acute Coronary Syndrome (NCVD-ACS) registry year 2006 to 2013. PATIENTS AND METHODS: Patients were categorized into four subgroups based on the year of admission (2006 to 2007, 2008 to 2009, 2010 to 2011 and 2012 to 2013). Baseline characteristics and clinical presentation, in-hospital pharmacotherapy, invasive revascularization and in-hospital/30-day mortality were analysed and compared between the subgroups. MAIN OUTCOME MEASURE(S): Rate of in-hospital catheterization/percutaneous coronary intervention. RESULTS: The registry contained data on 19483 patients. Intravenous thrombolysis was the main reperfusion therapy. Although the overall rate of in-hospital catheterisation/PCI more than doubled over the study period, while the use of primary PCI only slowly increased from 7.6% in 2006/2007 to 13.6% in 2012/2013. The use of evidence-based oral therapies increased steadily over the years except for ACe-inhibitors and angiotensin-receptor blockers. The adjusted risk ratios (RR) for in-hospital mortality for the four sub-groups have not shown any significant improvement. The 30-day adjusted risk ratios however showed a significant albeit gradual risk reduction (RR 0.773 95% CI 0.679-0.881, P < .001). CONCLUSION: Adherence to evidence-based treatment in STEMI in Malaysia is still poor especially in terms of the rate of primary PCI. Although there is a general trend toward reduced 30-day mortality, the reduction was only slight over the study period. Drastic effort is needed to improve adherence and clinical outcomes. LIMITATION: Retrospective registry data with inter-hospital variation.

Wedge Versus Core Biopsy at Time Zero: Which Provides Better Predictive Value for Delayed Graft Function With the Remuzzi Histological Scoring System?

BACKGROUND: Histopathological features on time-zero renal biopsies correlate with graft outcome after renal transplantation. With increasing numbers of marginal donors, assessment of pre-implantation graft quality is essential. The clinician's choice of wedge or core biopsy is performed without evidence of efficacy or safety. This study aims to compare the information derived from wedge biopsy versus core biopsy. METHODS: Prospective evaluation of 37 wedge biopsies and 30 core biopsies was performed. Histopathological data were collected on number of glomeruli and arterioles observed, and Remuzzi scoring for glomerulosclerosis, tubular atrophy, interstitial fibrosis, and arteriolar narrowing was performed. Clinical data on delayed graft function (DGF) were also collated. Sensitivity, specificity, and positive and negative predictive values for DGF were compared. RESULTS: Patient demographics between the two cohorts were comparable. No complications of biopsies occurred; 81% of wedge biopsies versus 50% of core biopsies had >10 glomeruli (P = .01), whereas 32% of wedge biopsies and 57% of core biopsies had >2 arterioles (P = .02). Wedge biopsies were more likely to identify pathology with more glomerulosclerosis, tubular atrophy (P < .01), and interstitial fibrosis (P < .01). There was a non-significant trend toward high Remuzzi scores in wedge biopsy (22% versus 7% with Remuzzi ≥ 4; P = .12). The sensitivity and positive predictive value of Remuzzi ≥ 4 for predicting DGF was better on wedge biopsy (45.5% versus 0%; P < .01 and 62.5% versus 0%; P < .01, respectively). CONCLUSIONS: Wedge biopsies were safe and superior to core biopsies for identifying clinically significant histopathological findings on pre-implantation renal biopsy. We believe that the wedge biopsy is the method of choice for time-zero biopsies.

Promotion of microfilament polymerization by depolymerized glia filament preparation through interaction of vimentin with actin.

We investigated the in vitro interaction between microfilaments and glia filaments in the presence of alpha-sialosylcholesterol (SC). The depolymerized glia filament preparation (GF) that had been extracted from a crude cytoskeletal fraction of cultured rat astrocytes promoted markedly the polymerization of the depolymerized microfilament preparation (MF) in the presence of SC. The ability of GF to promote the MF polymerization was highest in the concentration range 7.5-15 microM SC. The polymerization of MF at the concentration of 10 micrograms protein/200 microliters was enhanced to the highest level by 4 micrograms protein/200 microliters of GF in the presence of 15 microM SC. The MF polymerization was promoted more efficiently by GF than by MF: for example, the polymerization of actin in MF (7.5 micrograms/200 microliters) was enhanced by the addition of 5 micrograms GF protein/200 microliters to a twice greater extent than by the addition of 5 micrograms MF protein/200 microliters in the presence of 15 microM SC. GF enhanced the MF polymerization by over three-fold at 4-37 degrees C, and the GF activity became greater with an increase in incubation temperature. Vimentin promoted the G-actin polymerization in the presence of 11.3 microM alpha-SC. The finding suggested that the interaction between vimentin and actin was one of the factors to cause GF to promote the MF polymerization.

A monoclonal anti-HLA-B27 antibody which is reactive with a linear sequence of the HLA-B27 protein is useful for the study of molecular mimicry.

In the search for cross-reactivity between bacteria and HLA-B27, three groups of investigators have identified several bacterial envelope proteins which are reactive with the monoclonal anti-HLA-B27 antibodies B27.M1 and B27.M2. Since these two antibodies react poorly with HLA-B27-derived synthetic peptides, it is not possible to locate the reactive epitopes on the HLA-B27 using synthetic peptides. Here, we introduce Ye-2, a monoclonal anti-HLA-B27 antibody which, unlike B27.M1 and B27.M2, is reactive with a synthetic peptide derived from residues 63-84 of HLA-B27.1. Analysis with a cross-reactive peptide derived from residues 226-244 of bovine carbonic anhydrase suggests that only a few of the amino acid residues in the HLA-B27-derived peptide are responsible for the reactivity. This antibody should be a useful adjunct in a preliminary assessment of whether a bacterial protein mimics HLA-B27 in primary structure.

In vitro development of caprine ovarian preantral follicles.

The in vitro growth and developmental pattern of caprine preantral follicles cultured in agar gel was observed. Preantral follicles 50 to 150 microm in diameter were isolated from prepuberal goat ovaries by treatment with collagenase and DNase. The isolated preantral follicles were cultured in agar gel for up to 14 days. A group of 10 follicles in different developmental stages was cultured in a culture well coated with 0.6% agar gel and filled with DMEM medium supplemented with FCS (10%), hypoxanthine (2 mmol/mL), dbcAMP (2 mmol/mL), FSH (100 ng/mL), insulin-transferrin-selenium (ITS) (50 ng/mL), IGF-1 (50 ng/mL), hydrocortisone (40 ng/mL) and antibiotics. Follicle viability was determined under an inverted phase-contrast microscope according to morphological and histological criteria, and follicle growth was assessed by their size and appearance. The results showed that the three-dimensional structures and forms of follicles were basically maintained intact during culture. Primary follicles developed into secondary follicles and a few of them into antral follicles. A large portion of secondary follicles entered the antral stage, and oocytes also acquired growth. The formation of theca lamina and zona pellucida was observed. The survival capacity of secondary follicles was greater than primary follicles. The survival rates for primary and secondary follicles were 11.36% (5/44) and 71.16% (53/74), respectively. During in vitro development the follicles demonstrated dominance. This experiment revealed the preliminary characteristics of the in vitro development of caprine preantral follicles.

Immune activation of erythroleukemia cells induced by interleukin 12.

To investigate the antitumor activity of IL-12, the induction of differentiation of IL-12 was observed using erythroleukemia cells (FBL-3) as model. After incubation with 200 U/mL IL-12 for 48 h, DNA synthesis of FBL-3 cells in S phase decreased significantly; the expression of CD14 which is the specific marker of monocyte increased, the rate of NBT(+) cells was apparently higher than that of the untreated FBL-3 cells. After treating FBL-3 cells with IL-12 for 72 h, the expression of 33D1 and NLDC145 which are the specific markers of dendritic cells increased markedly, the surface molecules such as MHC-11, B7-1, B7-2, and VCAM-1 were up-regulated; morphological observation showed two kinds of cells: some cells had a ruffled surface and plentiful lysosome; the others had many dendritic projections on the surface, and contained numerous mitochondria. Functionally, the IL-12-treated FBL-3 cells could apparently stimulate the proliferation of allogeneic and autologous T lymphocytes, and improve the specific cytotoxic activity of CTL on FBL-3 cells. These results indicated that erythroleukemia cells were induced by IL12 to differentiate into the monocytes and dendritic cells, then exhibited the antigen-presenting function. The data outline a new mechanism for IL-12 to treat leukemia.

Effects of thienorphine on synaptic structure and synaptophysin expression in the rat nucleus accumbens.

The partial opioid agonist thienorphine is currently in Phase II clinical trials in China as a candidate drug for the treatment of opioid dependence. However, its effect on synaptic plasticity in the NAc (nucleus accumbens) remains unclear. In the present study, we measured structural parameters of the synaptic interface to investigate the effect of thienorphine, morphine or a combination of both on synaptic morphology in the NAc of rats. Expression of synaptophysin was also examined. Ultrastructural observation showed that synaptic alterations were less pronounced after chronic thienorphine administration than after chronic morphine administration. Animals that received thienorphine had thinner postsynaptic densities and shorter active zones in the NAc compared with those in the saline group, but the active zone was larger, and the cleft narrower, than those in the morphine group. Furthermore, synaptophysin expression in the NAc was significantly greater after chronic administration of thienorphine, morphine, or both, than after saline. These results identified interesting differences between thienorphine and morphine in their effects on synaptic structure and synaptophysin expression in the rat NAc. Further study is deserved to investigate thienorphine as a new treatment for opioid dependence.