Mannose is an important sugar in the biology of the Gram-negative bacterium Porphyromonas gingivalis. It is a major component of the oligosaccharides attached to the Arg-gingipain cysteine proteases, the repeating units of an acidic lipopolysaccharide (A-LPS), and the core regions of both types of LPS produced by the organism (O-LPS and A-LPS) and a reported extracellular polysaccharide (EPS) isolated from spent culture medium. The organism occurs at inflamed sites in periodontal tissues, where it is exposed to host glycoproteins rich in mannose, which may be substrates for the acquisition of mannose by P. gingivalis. Five potential mannosidases were identified in the P. gingivalis W83 genome that may play a role in mannose acquisition. Four mannosidases were characterized in this study: PG0032 was a β-mannosidase, whereas PG0902 and PG1712 were capable of hydrolyzing p-nitrophenyl α-d-mannopyranoside. PG1711 and PG1712 were α-1 → 3 and α-1 → 2 mannosidases, respectively. No enzyme function could be assigned to PG0973. α-1 → 6 mannobiose was not hydrolyzed by P. gingivalis W50. EPS present in the culture supernatant was shown to be identical to yeast mannan and a component of the medium used for culturing P. gingivalis and was resistant to hydrolysis by mannosidases. Synthesis of O-LPS and A-LPS and glycosylation of the gingipains appeared to be unaffected in all mutants. Thus, α- and β-mannosidases of P. gingivalis are not involved in the harnessing of mannan/mannose from the growth medium for these biosynthetic processes. P. gingivalis grown in chemically defined medium devoid of carbohydrate showed reduced α-mannosidase activity (25%), suggesting these enzymes are environmentally regulated.

The prostaglandin-evoked cAMP production was studied in human neuroblastoma SK-N-BE(2)C cells during neuronal differentiation induced by all-trans retinoic acid. The incubation with 5 microM all-trans retinoic acid for 4-6 days promoted neurite outgrowth of cells. After differentiation, prostaglandin E(2) (PGE(2))-induced cAMP production was dramatically increased, whereas forskolin- and AlF-induced cAMP productions were not changed. The increase reached maximum after 4-days of incubation with all-trans retinoic acid. The differentiation caused an increase in the maximal response and a decrease in the half-maximal effective concentration of the PGE(2)-induced cAMP production. In addition, the binding of [(3)H]PGE(2) to membrane receptors was enhanced in differentiated cells. However, the order of potency of the various prostaglandins (PGE(1) = PGE(2)>> PGD(2) = PGF(2alpha) = PGI(2)) in cAMP production did not change during the differentiation, suggesting that mainly E-prostanoid (EP) receptors were involved. Butaprost, an EP(2) receptor specific agonist, increased the cAMP level in a concentration dependent manner and had a similar potentiating effect on cAMP production as PGE(2) upon differentiation. Northern blot analysis using the human cDNA probes shows that the EP(2) mRNA level was about seven times higher in differentiated cells, while the dopamine beta-hydroxylase (DBH) mRNA completely disappeared. Our results, thus, suggest that elevated gene expression of the prostanoid EP(2) receptor results in an increase in the PGE(2)-evoked cAMP production in SK-N-BE(2)C cells during neuronal differentiation.

BACKGROUND

Superantigens and eicosanoids are important amplifiers and regulators of inflammation in airway diseases. We therefore studied the possible influence of Staphylococcus aureus enterotoxin B (SEB) on the cyclooxygenase (COX) pathway and basic functions of airway structural cells.

METHODS

Fibroblasts were isolated from nasal inferior turbinate tissue and cultured in the presence of different concentrations of SEB. Preincubation with interferon (IFN)-gamma was performed to induce expression of major histocompatibility complex (MHC) class II receptors. Prostaglandin E2 (PGE(2)) production was assayed by enzyme-linked immunosorbent assay, and levels of COX-2 and prostanoid E receptors 1-4 (EP(1-4)) were assayed by real-time polymerase chain reaction. Migration and growth tests were performed, and SEB was localized within the cells by confocal microscopy.

RESULTS

Stimulation with IFN-gamma and SEB significantly down-regulated PGE(2), COX-2, and EP(2) expression but not EP(1), EP(3), or EP(4) expression. The enterotoxin blocked cell growth but increased the fibroblast migration rate. SEB was localized within the cell in the presence and absence of MHC-II, suggesting that mechanisms other than conventional binding may allow the enterotoxin to enter the cell.

CONCLUSIONS

These findings may have major implications for our understanding of the role played by bacterial superantigens in regulating the inflammatory and remodeling mechanisms of upper airway diseases and hence may help elucidate the pathophysiology of these diseases.

Repeated injections of <em>Ep</em>-475, a potent cysteine proteinase inhibitor, into rats caused several-fold increase in the hepatic contents of the lysosomal cysteine proteinases cathepsin <em>B</em>, H and L and in the activities of other lysosomal hydrolases. The rates of degradation of these lysosomal enzymes, estimated by repeated injections of cycloheximide, were found to be retarded in <em>Ep</em>475-treated rats, indicating that lysosomal cysteine proteinases are involved in degradation of lysosomal enzymes including proteinases.

Real-time RT-PCR was used to quantify the expression of genes possibly involved in Mycobacterium tuberculosis latency in in vitro and murine models. Exponential and stationary phase (EP and SP) bacilli were exposed to decreasing pH levels (from 6.5 to 4.5) in an unstirred culture, and mRNA levels for 16S rRNA, sigma factors sigA,B,E,F,G,H and M, Rv0834c, icl, nirA, narG, fpbB, acr, rpoA, recA and cysH were quantified. The expression of acr was the one that best correlated with the CFU decrease observed in SP bacilli. In the murine model, the expressions of icl, acr and sigF tended to decrease when bacillary counts increased and vice versa. Values from immunodepressed mice (e.g. alpha/beta T cells, TNF, IFN-gamma and iNOs knock out strains), with accelerated bacillary growth rate, confirmed this fact. Finally, the expression of acr was maintained in mice following long-term treatment with antibiotics. The quantification of acr expression could be useful for monitoring the presence of latent bacilli in some murine models of tuberculosis.

A model for the regulation of erythropoietin production has been presented. This model proposes that a primary O2-sensing reaction in the kidney is initiated by a decrease in ambient PO2, a rapid decrease in gas exchange in the lung, a diminished oxygen-carrying capacity of hemoglobin, a molecular deprivation of oxygen, or a decrease in renal blood flow. It is proposed that the primary oxygen-sensing reaction may trigger the release of several mediators that stimulate adenylate cyclase through a receptor-activated stimulation of a G protein in the renal cell membrane. Some of the agents that are thought to be released during hypoxia, which may trigger this cascade, are adenosine (A2 activation), eicosanoids (PGE2, PGI2, and 6-keto PGE1), oxygen-free radicals (superoxide and H2O2), and catecholamines with beta-2 adrenergic receptor agonist properties. The activation of adenylate cyclase generates cyclic AMP, which activates protein kinase A, leading to the production of a phosphoprotein that, in turn, activates a nuclear protein involved in transcription and/or translation for erythropoietin biosynthesis and/or secretion. A second part of this model concerns the effect of hypoxia on a renal cell membrane phosphodiesterase and the generation of inositol triphosphate and diacylglycerol. Diacylglycerol may interact with diacylglycerol lipase to generate arachidonic acid, which, together with arachidonic acid generated by the interaction of phospholipase A2 on membrane phospholipids, produces eicosanoids. Eicosanoids may play a secondary role in Ep production/secretion. The model further proposes that calcium levels in both renal and liver cells may be important in regulating erythropoietin biosynthesis and/or secretion. It is proposed that an increase in intracellular calcium leads to the inhibition of erythropoietin biosynthesis and/or secretion and a decrease in intracellular calcium increases erythropoietin production. The specific mechanism by which calcium regulates erythropoietin biosynthesis and secretion is not well understood. However, a good correlation is seen with several agents that decrease intracellular calcium and increase erythropoietin production as well as with other agents that increase intracellular calcium and decrease erythropoietin production. When inositol triphosphate levels are increased, an increase in the mobilization of intracellular calcium from the endoplasmic reticulum or another intracellular pool occurs. This increased intracellular calcium probably activates a calcium calmodulin kinase and produces a phosphoprotein that inhibits erythropoietin production/secretion.(ABSTRACT TRUNCATED AT 400 WORDS)

Hermansky-Pudlak syndrome (HPS) is a hereditary disorder involving the sorting processes of intracellular organelles such as lysosomes of reticuloendothelial cells. Pale ear (ep) mouse is known to have the HPS1 gene mutation, which is seen in patients with HPS and pulmonary fibrosis. As pulmonary fibrosis is not spontaneously observed in ep mice, we hypothesized that external stimuli are necessary for the genetic predisposition of its development. We used silica as the external stimulus to induce the alveolar macrophage-mediated inflammatory response and evaluated the pathological changes of the lung and biochemical analysis of collagenolytic lysosomal enzymes cathepsins L and B in ep mice. Treatment with silica induced the following: persistent accumulation of activated macrophages; delayed clearance of silica from alveolar spaces; and increased collagen fibers in alveolar tissues, which were shown with trichrome staining in ep mice. The comparison of bronchoalveolar lavage cells between the naïve ep and control mice revealed: decreased enzymatic activities but increased antigenic levels of cathepsins L and B, resulting in significantly lower ratios of activity to antigen; increased ceroid deposits and cathepsin L antigens in lysosomes; and no abnormal forms of cathepsins were detected. After silica instillation, activities of cathepsin L in the ep mice increased but ratios of activity to antigen were still significantly low. These phenomena induced by silica suggest that external stimuli bring forth fibrogenesis in the animal models or humans that have HPS1 gene mutation.

Age-dependency has been demonstrated for classic diastolic parameters but is less documented for novel markers. We compared wide spectrum of diastolic measurements in healthy subjects younger and older than 50 and 70 years.

METHODS

In 80 subjects (17 to 91 year, 38 male, mean age 53 +/- 16) we assessed in transthoracic echocardiography mitral inflow, pulmonary vein flow, propagation velocity, and tissue Doppler parameters. The data were compared between the groups: A < 50, B 50-70, and C>> 70 years and their correlation with age was analyzed.

RESULTS

In the older groups a longer duration of isovolumic relaxation, higher velocity of mitral inflow atrial phase, and lower early to atrial mitral inflow velocity ratio (E/A) were observed. In pulmonary veins ratio of peak systolic to diastolic velocity (S/D) was higher and atrial reversal flow faster. Among the new parameters in older groups a lower propagation velocity of early wave (Ep) (30 +/- 8 in C, 44 +/- 10 in B vs 49+/-9 cm/s in A;P< 0.05) and peak early diastolic velocity of mitral annulus (E' 9.5 +/- 1.8 in C and 11.2 +/- 2.9 in B vs 15.8 +/- 4.1 cm/s in A,P< 0.05) were recorded.

CONCLUSIONS

Besides the evolution of mitral inflow toward the delayed relaxation profile in elderly, a decrease of propagation and tissue Doppler velocities were documented. The strongest positive correlation with age was observed for atrial velocity of mitral inflow (A, r = 0.63) and inverse correlation for E/A ratio (r =-0.6) and early diastolic velocity of mitral annulus (r =-0.69).

A peanut sprout is known to contain a significant level of resveratrol, which was reported to have beneficial effects in our body due to its antioxidant activities. The purpose of this study was to evaluate the cytoprotective activity of ethanol extract of peanut sprout (EPS) from ultraviolet B (UVB)-induced oxidative stress in human dermal fibroblasts (HDF). EPS was revealed to contain 54.2 μg g(-1) of trans-resveratrol. The DCF-DA-positive reactive oxygen species level was increased by 50 mJ cm(-2) of UVB irradiation (2150 ± 450% of nonirradiated control), which was markedly suppressed by EPS treatment (180 ± 42% of control). Annexin V-positive apoptotic cell death induced by UVB irradiation (16.4 ± 4.5%) was also significantly inhibited by EPS treatment (6.7 ± 2.5%). EPS induced up-regulation and nuclear translocation of Nrf2, a transcription factor for antioxidant and detoxifying enzymes, in HDF as a dose-dependent manner. UVB irradiation up-regulated Nrf2-dependent enzymes of heme oxygenase-1, NAD(P)H:quinine oxidoreductase-1 and glutathione-S-transferase pi, and they were further stimulated by EPS treatment. Taken together, EPS is an efficient cytoprotective agent against UVB-induced oxidative stress by activation of Nrf2 and upregulation of Nrf2-relating antioxidant and detoxifying enzymes in HDF.

The major objective of this study was to investigate the analogy existing between the typical circadian periodicity of ACTH and that recently described of beta-lipotropin (beta-LPH) and beta-endorphin (beta-EP) plasma levels. The determination of their concentrations, plus cortisol, has been performed on the same plasma samples of 6 healthy volunteers. All hormones were measured by radioimmunoassay. Those of beta-LPH and beta-EP were preceded by a purification of plasma through silicic acid extraction and Sephadex G-75 gel filtration. The highest values (mean +/- SEM) were found in the morning (ACTH: 10.3 +/- 0.9; beta-LPH; 6.3 +/- 0.7; beta-EP: 6.5 +/- 0.5 fmol/ml; cortisol: 378 +/- 30 pmol/ml) and the lowest values in the evening (ACTH: 6:1 +/- 0.7; beta-LPH: 3.3 +/- 0.4; beta-EP: 3.7 +/- 0.6 fmol/ml; cortisol: 130 +/- 23 pmol/ml). Statistical analysis using the Fourier method led to the evidence of a concomitant circadian secretory pattern of the three proopiocortin-related peptides. These results strongly suggest that the phasic secretion of ACTH, beta-LPH and beta-EP underlies a common central control.

OBJECTIVE

Aldosterone antagonists (AldoAs) have been used to treat severe chronic heart failure (CHF). There is uncertainty regarding the efficacy of using AldoAs in mild to moderate CHF with New York Heart Association (NYHA) classifications of I to II. This study summarizes the evidence for the efficacy of spironolactone (SP), eplerenone (EP) and canrenone in mild to moderate CHF patients.

METHODS

PubMed, MEDLINE, EMBASE and OVID databases were searched before June 2012 for randomized and quasi-randomized controlled trials assessing AldoA treatment in CHF patients with NYHA classes I to II. Data concerning the study's design, patients' characteristics and outcomes were extracted. Risk ratio (RR) and weighted mean differences (WMD) or standardized mean difference were calculated using either fixed or random effects models.

RESULTS

Eight trials involving 3929 CHF patients were included. AldoAs were superior to the control in all cause mortality (RR 0.79, 95% CI 0.66, 0.95) and in re-hospitalization for cardiac causes (RR 0.62, 95% CI 0.52, 0.74), the left ventricular ejection fraction was improved by AldoA treatment (WMD 2.94%, P = 0.52). Moreover, AldoA therapy decreased the left ventricular end-diastolic volume (WMD -14.04 ml, P < 0.00001), the left ventricular end-systolic volume (WMD -14.09 ml, P < 0.00001). A stratified analysis showed a statistical superiority in the benefits of SP over EP in reducing LVEDV and LVESV. AldoAs reduced B-type natriuretic peptide concentrations (WMD -37.76 pg ml(-1), P < 0.00001), increased serum creatinine (WMD 8.69 μmol l(-1), P = 0.0003) and occurrence of hyperkalaemia (RR 1.78, 95% CI 1.43, 2.23).

CONCLUSIONS

Additional use of AldoAs in CHF patients may decrease mortality and re-hospitalization for cardiac reasons, improve cardiac function and simultaneously ameliorate LV reverse remodelling.

BACKGROUND

Tacrolimus (Tac) is a macrolide immunosuppressant drug isolated from Streptomyces tsukubaensis, widely used in organ transplantation.

OBJECTIVE

This study examined the effect of tacrolimus administered by oral route (p.o.) on inflammation in mouse subcutaneous air pouch triggered by carrageenan (Cg 1%).

METHODS

The air pouch was induced as described by Benincá et al. [Benincá JP, Montanher AB, Zucolotto SM, Schenkel EP, FrödeTS. Anti-inflammatory effects of the Passiflora edulis: forma flavicarpa Degener inhibition of leukocytes, enzymes and pro-inflammatory cytokine levels in the air pouch model, in mice. Food Chem 2007; 104(3); 1097-1105.]. The inflammatory parameters (leukocytes, exudation, myeloperoxidase (MPO) and adenosine-deaminase (ADA) activities, as well as nitrate/nitrate concentrations (NO(x)), interleukin-1 beta (IL-1beta), chemokine to neutrophil (KC) and tumor necrosis factor-alpha (TNF-alpha) levels were analysed 24 h after injection of carrageenan.

RESULTS

Tacrolimus, indomethacin and dexamethasone significantly inhibited leukocytes, neutrophils and exudation (P<0.05) when they were administered 0.5 h before inflammation. These drugs, under the same conditions, decreased MPO and ADA activities (P<0.05), NO(x) and IL-1beta levels (P<0.01). Tacrolimus and indomethacin, but not dexamethasone, inhibited KC levels (P<0.01). On the other hand, tacrolimus and dexamethasone, but not indomethacin, decreased TNF-alpha levels (P<0.01).

CONCLUSIONS

Results of this study indicate that tacrolimus has an important anti-inflammatory property, showing not only inhibition of pro-inflammatory mediators release, but also inhibition of activated leukocyte infiltration into the site of inflammation. Furthermore, these results showed that most of the anti-inflammatory actions of tacrolimus were similar to those observed in animals treated with either indomethacin or dexamethasone.

OBJECTIVE

We examined ethnic and language disparities in diabetes care and management among California residents with type 2 diabetes based on ethnicity and English-language proficiency.

METHODS

Data were drawn from the 2007 California Health Interview Survey with a total of 3,531 Asian, Latino, and Caucasian adults with diabetes. Latino and Asian groups were subdivided by their limited English proficiency (LEP) level. Population-weighted regression analyses were conducted to examine group differences, controlling for socioeconomic and clinical variables.

RESULTS

Latino English Proficient (EP) and Latino LEP respondents received fewer hemoglobin A1c checks (EP: b = -0.11, P < .05; LEP: b = -0.27, P < .01) than Caucasians. Latino and Asian LEP respondents checked their glucose less frequently than Caucasians (Latino LEP: b = -0.49, P < .05; Asian LEP: b = -0.79, P < .01). Asian LEP respondents were less likely to receive feet checks than Caucasians (Asian LEP: b = -.52, P < .001). Asian LEP respondents received significantly fewer feet checks than Asian EP respondents (P < .05).

CONCLUSIONS

Ethnic disparities in disease management exist among California residents. However, beyond ethnicity, English proficiency should be taken into account when examining diabetes management among minority groups. Diabetic Californians who belong to ethnic minorities and speak limited English, particularly Asians, are less likely to receive the standard of care for diabetes than English proficient Caucasians. From a policy perspective, care should be taken to ensure that adequate information about diabetes management is available in multiple languages for patients with limited English skills.

Neuroendocrine and sympathoadrenal responses to exhaustive graded treadmill exercise were examined in 17 male subjects of varying degrees of fitness. The mean duration of exercise to exhaustion was 15.2 +/- 0.7 (+/- SE) min. Exercise duration was inversely correlated with baseline heart rate (P less than 0.05). Compared to standing baseline values, mean plasma norepinephrine and epinephrine levels increased 339% and 301%, respectively, in an integrated 2-min blood sample collected immediately after completion of exercise. Mean adrenocorticotrophic hormone (ACTH), beta-endorphin (beta-EP), beta-lipotropin (beta-LPH), and prolactin levels increased 282%, 720%, 372%, and 211%, respectively, in an integrated 4-min blood sample beginning 2 min after completion of exercise. Cortisol levels increased 183% in the sample collected 17-21 min after exercise. The magnitude of these neuroendocrine responses to exercise was similar among individuals at the same relative intensity of exhaustive exercise, regardless of the duration of exercise. The exercise-induced increases of the pro-opiomelanocortin (POMC)-derived peptides, ACTH, beta-EP, and beta-LPH, were highly correlated with each other (P values less than 0.001), and were correlated with prolactin increases, (P values less than 0.05). During a 20-min recovery period after exercise, changes in heart rate, ACTH, and beta-LPH levels were correlated with duration of exercise, (P less than 0.01, P less than 0.03, and P less than 0.03, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

OBJECTIVE

To evaluate the usefulness of a modified technique of endoscopic papillectomy (EP) for lessening the occurrence of complications.

METHODS

Indications for EP were adenoma or well-differentiated adenocarcinoma confined to the papilla of Vater (T1) without tumor spread into the bile/pancreatic duct. Sixteen patients underwent the modified technique, which consists of resection with the Endocut mode, followed by biliary/pancreatic sphincterotomy and stenting (Group A). Twelve patients who had undergone EP, using a cutting current, followed by pancreatic duct stenting were included as control (Group B). The frequency of complications and clinical outcomes were retrospectively compared between the two groups.

RESULTS

Sixteen patients had adenoma, and 12 had adenocarcinoma. Early complications occurred in 36% of all patients (hemorrhage, 7; cholangitis, 3; perforation, 2; cholecystitis, 1). The frequency of early complications in Group A was significantly lower than that in Group B (6% vs 75%, odds ratio [OR] 0.022, 95% confidence interval [CI], 0.0020-0.25). Late complications occurred in 18% of the patients (bile duct stone, 3; hemorrhage, 1; pancreatitis, 1). There was no significant difference in the late complication rate between Group A and Group B (19% vs 17%). Local recurrences were found in 14% of the patients, without a significant difference between Group A and Group B (19% vs 8%) during a mean follow-up period. All recurrent tumors were successfully treated with argon plasma coagulation.

CONCLUSIONS

The modified technique of EP for ampullary neoplasm contributed to lessening the occurrence of early complications. However, further refinement of this technique is necessary for improving the clinical outcome.

The equilibria among the central complexes in the phosphoglucomutase system were evaluated by (a) using an excess of enzyme plus Mg2+ to prepare mixtures with glucose phosphates in which essentially no free glucose phosphates were present; (b) inactivating the enzyme in such mixtures by means of a procedure that prevents substantial interconversion of the central complexes; and (c) assaying the quenched mixture for glucose 1-P, glucose 1-6-P2, and glucose-6-P. The fractional amounts of Ep-Mg-Glc-1-P, ED-Mg-Glc-P2, and Ep-Mg-Glc-6-P present at pH 7.5 and 24 degrees C were 0.13, 0.54, and 0.33. (Ep and ED are the phospho and dephospho forms of the enzyme, respectively). From these fractions and the equilibrium isotope exchange constants for the three sugar phosphates, true dissociation constants can be calculated for each of the above complexes: 8.5 muM, 19 nM, and 57 muM, respectively. Relative to the rate of PO3 transfer to water, a 3 x 10(10)-fold rate increase is produced by binding glucose-1-P to the Mg2+-enzyme (Ray, jr., W.J., Long, J.W., and Owens, J.D. (1976), Biochemistry, the following paper in this issue). This "substrate-induced rate effect" is equivalent to a difference of some 14 kcal in Gibbs activation energies for transfer to chemically similar hydroxyl groups, and most of this energy difference ultimately must be rationalized in terms of binding interactions involving the phosphoglucosyl moiety. Three different mechanisms for using substrate binding energy to reduce the activation energy of the subsequent catalytic step are examined as possible explanations for the substrate-induced rate effect. These mechanisms emphasize (a) enthalpic destabilization and (b) (entropic) immobilization of reactant groups during formation of the enzyme-substrate complex, and (c) increased binding interactions of nonreactant groups during the subsequent approach to the transition state. As a test for enthalpic destabilization of the enzymic phosphate group, values of deltaG degrees' for the hydrolytic cleavage of this group in Ep and Ep-Glc-1-P are calculated from equilibria measured at pH 7.5 and 30 degrees C: about -1 and +1.4 kcal/mol, respectively. To test for destabilization of the acceptor hydroxyl group in the enzyme-substrate complex, deltaG degrees' for equilibrium, Ep-Glc-P in equilibrium ED-Glc-P2, is compared with that for the corresponding process involving the nonrigid acceptor, 1,4-butanediol monophosphate: about -0.9 and -1.9 kcal, respectively. These results are not consistent with a large enthalpic destabilization of the reactant groups in the Ep-Glc-1-P complex. To test for entropic immobilization of reactant groups, glucose-6-phosphate is considered as a bidentate ligand, and the chelate effect on the binding and subsequent enzymic transfer reaction that arises from covalently linked the sugar ring and the PO3 group is evaluated. Reference reactions involving xylose as a PO3 acceptor both in the presence and absence of bond (inorganic) phosphite are used...

Yunnan has the greatest share of reported human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) cases in China. In recent years, HIV prevalence and incidence remained stubbornly high in men who have sex with men (MSM). To follow the dynamics of the HIV-1 epidemic among MSM, HIV-1 genetic characteristics and genetic transmission networks were investigated.

Blood samples from 190 newly diagnosed HIV-1 cases among MSM were continuously collected at fixed sites from January 2013 to December 2015 in Kunming City, Yunnan Province. Partial gag, pol and env genes were sequenced and used for phylogenetic and genotypic drug resistance analyses. The genetic characteristics of the predominant HIV-1 strains were analyzed by the Bayesian Markov Chain Monte Carlo (MCMC) method. The genetic transmission networks were identified with a genetic distance of 0.03 substitutions/site and 90% bootstrap support.

Among the 190 HIV-1 positive MSM reported during 2013-2105, various genotypes were identified, including CRF01_AE (45.3%), CRF07_BC (35.8%), unique recombinant forms (URFs) (11.6%), CRF08_BC (3.2%), CRF55_01B (2.1%), subtype B (1.6%) and CRF59_01B (0.5%). The effective population sizes (EPS) for CRF01_AE and CRF07_BC increased exponentially from approximately 2001-2010 and 2005-2009, respectively. Genetic transmission networks were constructed with 308 pol sequences from MSM diagnosed during 2010-2015. Of the 308 MSM, 109 (35.4%) were identified in 38 distinct clusters. Having multiple male partners was associated with a high probability of identification in the genetic transmission networks. Of the 38 clusters, 27 (71.1%) contained individuals diagnosed in different years. Of the 109 individuals in the networks, 26 (23.9%) had ≥2 potential transmission partners (≥2 links). The proportion of MSM with ≥2 links was higher among those diagnosed from 2010-2012. The constituent ratios of their potential transmission partners by areas showed no significant difference among MSM from Kunming, other cities in Yunnan and other provinces. Additionally, surveillance drug resistance mutations (SDRMs) were identified in 5% of individuals.

This study revealed the various HIV-a genotypes circulating among MSM in Kunming. MSM with more partners were more easily detected in transmission networks, and early-diagnosed MSM remained active in transmission networks. These findings suggested that the routine interventions should be combined with HIV testing and linkage to care and early antiretroviral therapy among HIV-positive MSM.

Multiple human immunodeficiency virus (HIV)-1 genotypes in China were first discovered in Yunnan Province before disseminating throughout the country. As the HIV-1 epidemic continues to expand in Yunnan, genetic characteristics and transmitted drug resistance (TDR) should be further investigated among the recently infected population. Among 2828 HIV-positive samples newly reported in the first quarter of 2014, 347 were identified as recent infections with BED-captured enzyme immunoassay (CEIA). Of them, 291 were successfully genotyped and identified as circulating recombinant form (CRF)08_BC (47.4%), unique recombinant forms (URFs) (18.2%), CRF01_AE (15.8%), CRF07_BC (14.4%), subtype C (2.7%), CRF55_01B (0.7%), subtype B (0.3%) and CRF64_BC (0.3%). CRF08_BC and CRF01_AE were the predominant genotypes among heterosexual and homosexual infections, respectively. CRF08_BC, URFs, CRF01_AE and CRF07_BC expanded with higher prevalence in central and eastern Yunnan. The recent common ancestor of CRF01_AE, CRF07_BC and CRF08_BC dated back to 1983.1, 1992.1 and 1989.5, respectively. The effective population sizes (EPS) for CRF01_AE and CRF07_BC increased exponentially during 1991-1999 and 1994-1999, respectively. The EPS for CRF08_BC underwent two exponential growth phases in 1994-1998 and 2001-2002. Lastly, TDR-associated mutations were identified in 1.8% of individuals. These findings not only enhance our understanding of HIV-1 evolution in Yunnan but also have implications for vaccine design and patient management strategies.

Previous work has suggested that the antinociceptive effect of nitrous oxide (N2O) in rats is mediated, at least in part, by beta-endorphin (beta-EP) and that centrally administered beta-EP stimulates release of methionine-enkephalin (ME) in the rat spinal cord. Since inhibition of central nitric oxide (NO) production has been found to suppress N2O antinociception, we examined the possible involvement of NO in the release of spinal cord ME by i.c.v. beta-EP. Urethane-anesthetized, male Sprague-Dawley rats were intrathecally (i.t.) perfused with artificial cerebrospinal fluid (aCSF) and fractions of perfusate were assayed for immunoreactive (i.r.) ME. The beta-EP-induced increase in ME concentration in the i.t. perfusate was significantly suppressed by perfusing the animal with aCSF containing 100 microM L-NG-nitro arginine (L-NOARG), an inhibitor of NO synthase (NOS). The further addition of 50 microM L-arginine (L-ARG), but not D-arginine (D-ARG), to the aCSF reversed the suppression of the ME change by L-NOARG. However, the potency of L-ARG decreased with increasing concentrations of L-ARG. On the other hand, increasing the concentration of L-NOARG in the aCSF to 250 microM failed to produce a greater suppression of the beta-EP-induced increase in ME. These findings suggest that NO may mediate the beta-EP-induced release of ME in the spinal cord and that interference with this mechanism might be an explanation for the antagonism of N2O antinociception in rats by NOS inhibitors.

The cysteine endoproteases (EP)-A and EP-B were purified from green barley (Hordeum vulgare L.) malt, and their identity was confirmed by N-terminal amino acid sequencing. EP-B cleavage sites in recombinant type-C hordein were determined by N-terminal amino acid sequencing of the cleavage products, and were used to design internally quenched, fluorogenic peptide substrates. Tetrapeptide substrates of the general formula 2-aminobenzoyl-P2-P1-P1'-P2'-tyrosine(NO2)-aspartic acid, in which cleavage occurs between P1 and P1', showed that the cysteine EPs preferred phenylalanine, leucine, or valine at P2. Arginine was preferred to glutamine at P1, whereas proline at P2, P1, or P1' greatly reduced substrate kinetic specificity. Enzyme cleavage of C hordein was mainly determined by the primary sequence at the cleavage site, because elongation of substrates, based on the C hordein sequence, did not make them more suitable substrates. Site-directed mutagenesis of C hordein, in which serine or proline replaced leucine, destroyed primary cleavage sites. EP-A and EP-B were both more active than papain, mostly because of their much lower Km values.

Procedures have been developed to label human erythropoietin (Ep) with biotin to detect and isolate the Ep-receptor. The labeling method used the abundant carbohydrate groups on Ep and resulted in biologically active biotin-Ep (b-Ep) containing 8 to 10 biotins per Ep molecule. Specific binding of b-Ep to cells from spleens of mice made anemic by phenylhydrazine injections was demonstrated using 125I-labeled streptavidin. B-Ep, together with fluorescently tagged streptavidin, was found to specifically detect Ep-receptor-bearing cells by flow cytometry. This was demonstrated in several ways. First, approximately 90% of nucleated spleen cells from phenylhydrazine-treated mice were clearly fluorescent after staining with b-Ep and streptavidin-phycoerythrin, whereas only background fluorescence was detected using spleen cells from untreated mice. In addition, Ep-receptors were detected on 5% to 10% of normal mouse bone marrow cells, and these cells could be identified as erythroid in nature by separating the cells into subpopulations based on light-scatter properties. Third, Ep-receptor expression was found to correlate positively with expression of transferrin receptors, confirming the erythroid nature of these cells. B-Ep was also used to isolate Ep-receptors from monkey COS cells transfected with the murine Ep-receptor cDNA. In these experiments a cell-surface-bound protein of approximately 65 Kd and an intracellular protein of approximately 60 Kd were isolated from these cells. The procedures described in this report for detecting Ep-receptor expressing cells and for isolating the Ep-receptor should be valuable for purifying erythroid cells from heterogeneous cell populations, for elucidating the structure of the Ep-receptor, and for studying the biological activities of Ep at the cellular and molecular level.

Propionibacterium freudenreichii is a bacterial species found in Swiss-type cheeses and is also considered for its health properties. The main claimed effect is the bifidogenic property. Some strains were shown recently to display other interesting probiotic potentialities such as anti-inflammatory properties. About 30% of strains were shown to produce a surface exopolysaccharide (EPS) composed of (1→3,1→2)-β-D-glucan due to a single gene named gtfF. We hypothesized that functional properties of P. freudenreichii strains, including their anti-inflammatory properties, could be linked to the presence of β-glucan. To evaluate this hypothesis, gtfF genes of three β-glucan-producing strains were disrupted. These knockout (KO) mutants were complemented with a plasmid harboring gtfF (KO-C mutants). The absence of β-glucan in KO mutants was verified by immunological detection and transmission electron microscopy. We observed by atomic force microscopy that the absence of β-glucan in the KO mutant dramatically changed the cell's topography. The capacity to adhere to polystyrene surface was increased for the KO mutants compared to wild-type (WT) strains. Anti-inflammatory properties of WT strains and mutants were analyzed by stimulation of human peripheral blood mononuclear cells (PBMCs). A significant increase of the anti-inflammatory interleukin-10 cytokine production by PBMCs was measured in the KO mutants compared to WT strains. For one strain, the role of β-glucan in mice gut persistence was assessed, and no significant difference was observed between the WT strain and its KO mutant. Thus, β-glucan appears to partly hide the anti-inflammatory properties of P. freudenreichii; which is an important result for the selection of probiotic strains.

As part of environmental management policies in Europe, separate collection of organic household waste and nonorganic household waste has become increasingly common. As waste is often stored indoors, this policy might increase microbial exposure in the home environment. In this study we evaluated the association between indoor storage of organic waste and levels of microbial agents in house dust. The levels of bacterial endotoxins, mold beta(1-->3)-glucans, and fungal extracellular polysaccharides (EPS) of Aspergillus and Penicillium species were determined in house dust extracts as markers of microbial exposure. House dust samples were collected in 99 homes in The Netherlands selected on the basis of whether separated organic waste was present in the house. In homes in which separated organic waste was stored indoors for 1 week or more the levels of endotoxin, EPS, and glucan were 3.2-, 7.6-, and 4. 6-fold higher, respectively (all P < 0.05), on both living room and kitchen floors than the levels in homes in which only nonorganic residual waste was stored indoors. Increased levels of endotoxin and EPS were observed, 2.6- and 2.1-fold (P < 0.1), respectively, when separated organic waste was stored indoors for 1 week or less, whereas storage of nonseparated waste indoors had no effect on microbial agent levels (P>> 0.2). The presence of textile floor covering was another major determinant of microbial levels (P < 0.05). Our results indicate that increased microbial contaminant levels in homes are associated with indoor storage of separated organic waste. These increased levels might increase the risk of bioaerosol-related respiratory symptoms in susceptible people.

OBJECTIVE

To investigate the diagnostic relevance of serum cytokine concentrations in ectopic pregnancy (EP).

METHODS

Cohort study.

METHODS

University hospital.

METHODS

Seventeen women with EP, 22 women with miscarriage, and 33 women with normal intrauterine pregnancy, at comparable stages of gestation.

METHODS

Interleukin (IL)-2 receptor, IL-6, IL-8, and tumor necrosis factor alpha (TNF-alpha) determination by immunoradiometric assay.

METHODS

Serum concentrations of progesterone, beta-hCG, IL-2R, IL-6, IL-8, and TNF-alpha.

RESULTS

Serum levels of IL-6 were higher in women with EP than in those with miscarriage and normal pregnancy. Serum levels of TNF-alpha were higher in women with EP than in those with miscarriage and normal pregnancy. Serum levels of IL-8 were higher in women with EP than in those with miscarriage and normal pregnancy. An IL-8 cutoff of >40 pg/mL predicted EP with a sensitivity of 82.4%, a specificity of 81.8%, and positive and negative predictive values of 58.3% and 93.8%. No difference in serum IL-2R levels was found among the groups.

CONCLUSIONS

Serum IL-8, IL-6, and TNF-alpha concentrations are higher in women with EP than in those with miscarriage and normal pregnancy. Further studies are needed to determine their diagnostic value.