Evaluating Potential Vaccine Antigens in both the <i>Chlamydia trachomatis</i>and <i>Chlamydia muridarum</i>Intravaginal Mouse Challenge Models

Identifying relevant animal challenge models adds to the complexity of human vaccine development. Murine challenge models have been the most utilized animal model for Chlamydia trachomatis vaccine development. The question arises as to whether the C. trachomatis or C. muridarum pre-clinical model is optimal. We compared C. muridarum and C. trachomatis intravaginal challenge models in a combined total of seventy-five studies evaluating potential vaccine candidates. In 100% (42/42) of C. muridarum studies, mice immunized with Chlamydia elementary bodies (EB) demonstrated a significant reduction in urogenital bacterial shedding as measured by qPCR (p C. trachomatis studies. We have evaluated proposed vaccine antigens in both models and observed immunization with Chlamydia major outer membrane protein (MOMP) vaccine formulations to be protective (p C. trachomatis model, and immunization with PmpD p82 translocator domain was not protective in either model. We also observed in both models that depletion of CD4+ T-cells in MOMP-immunized mice resulted in diminished protective immunity but animals were still able to reduce the infection level. In contrast, mice immunized with live EBs by intraperitoneal route did not require CD4+ T-cells to resolve urogenital infection from intravaginal challenge in either model. Overall, we have found the C. muridarum model to be a more robust, reliable, and reproducible model for vaccine antigen discovery.

Safety and Efficacy of Combination Therapy with Insulin Glargine and Oral Hypoglycaemic Agents Including DPP-4 Inhibitors in Japanese T2DM Patients: ALOHA 2 Study, a Post-Marketing Surveillance for Lantus^&reg;

Aims: In the Add-on Lantus&reg;?to Oral Hypoglycaemic Agents 2 (ALOHA 2) Study in Japanese adults with type 2 diabetes mellitus (T2DM), data on the safety and efficacy of combination therapy with insulin glargine (Lantus&reg;) and oral anti-hyperglycaemic drugs (OADs) including dipeptidyl peptidase-4 (DPP-4) inhibitors in a real-life setting were collected and analyzed. Methods: This postmarketing surveillance was a prospective, observational, 24-week study that complied with the pharmaceutical affairs law and the ministerial ordinance of “Good Post-Marketing Study Practice (GPSP)” in Japan. Safety, efficacy and patient-reported outcomes (PROs);patients’ satisfaction with treatment (DTSQs and DTSQc) and patients’ self-reported health (EQ-5D and EQ-VAS) of combination therapy of insulin glargine and OADs were evaluated. Results: A total of 2,630 patients were enrolled. Of the 2,602 patients in the safety analysis population, 161 patients experienced 175 cases of adverse drug reactions, and the major adverse drug reaction was hypoglycaemia (140 patients, 5.38%). Out of those with hypoglycaemia, 11 patients (0.42%) had severe hypoglycaemia and the incidence rate (episodes per patient-year) was 0.019. Basal supported oral therapy (BOT) with insulin glargine substantially reduced the HbA1c, FPG and 2 hour-PPG levels for 24 weeks by -1.61%, -54.4 mg/dL and -74.5 mg/dL respectively. The mean weight was increased, however the change was +0.50 kg. In addition, the treatment satisfaction scores of DTSQs (mean treatment satisfaction score increased 3.6 from baseline to last observation) and DTSQc, EQ-5D index scores and EQ-VAS scores were significantly improved. Conclusion: Insulin glargine and OADs combination therapy was suggested to be effective and well tolerated. Patients’ satisfaction with treatment and their self-reported health improved in spite of the addition of injections to oral agents. The combination therapy of insulin glargine and OADs including DPP-4 inhibitors is likely to be considered an important therapeutic option in the diabetic patients.

Evaluation of previous product residue on magnetic bar used for manufacturing of Japanese Encephalitis vaccine

Cleaning validation is one of recommendation from Pharmaceutical Inspection Co-operation Scheme(PIC/S)to avoid contamination of pharmaceutical products.The objective of cleaning validation is to confirm reliability of cleaning procedure for removal of contaminants associated with previous products,residues of cleaning agents and potential microbial contaminants(1)Japanese Encephalitis vaccine is a new product that is prequalified by World Health Organization(WHO)and manufactured at GPO-MBP.

The Burden of Hospitalisations for Herpes Zoster Ophtalmicus in Spain (1997-2008)

Purpose: This epidemiological retrospective survey aims to provide population-based estimates of the burden of hospitalisation for herpes zoster ophthalmicus in Spain during a twelve-year period (1997-2008). Methods: All hospital discharges and deaths related to herpes zoster ophthalmicus in patients older than 30 years old from 1997 to 2008 in Spain were obtained. Differences in proportions were assessed by the Chi-square test. ANOVA was used for multiple comparisons. Poisson regression was used to assess differences in the rates. Results: A total of 3029 hospital discharges in patients older than 30 years old were coded as herpes zoster ophthalmicus. Of those, 1009 were coded as primary cause of hospitalization. This corresponds to a hospitalization rate of 0.94 and 0.31 hospitalizations per 100,000 for any position and first diagnostic position, respectively. The 75% of the total discharges were in immunocompetent patients. The herpes zoster ophthalmicus hospitalization rate increased significantly during the study period. This increase was also observed for immunocompetent patients. A total of 127 and 21 deaths occurred in patients ≥ 30 year-old hospitalized with herpes zoster ophthalmicus in any diagnostic position and principal diagnostic code, respectively during the 12-year study period. Of them, 70 (53.4%) were in immunocompetent patients, and, of those 11 were in patients with HZO in the first diagnosis position. The hospitalization rate and case-fatality rate increase significantly with age from 50 years old onwards. Conclusion: HZO, a vaccine preventable disease, still causes hospitalization and death in adults in Spain.

Meaningful Contact Estimates among Children in a Childcare Centre with Applications to Contact Matrices in Infectious Disease Modelling

We present a mathematical model of a day care center in a developed country (such as Canada), in order to use it for the estimation of individual-to-individual contact rates in young age groups and in an educational group setting. In our model, individuals in the population are children (ages 1.5 to 4 years) and staff, and their interactions are modelled explicitly: person-to-person and person-to-environment, with a very high time resolution. Their movement and meaningful contact patterns are simulated and then calibrated with collected data from a child care facility as a case study. We present these calibration results as a first part in the further development of our model for testing and estimating the spread of infectious diseases within child care centers.

Widening socioeconomic disparity in lung cancer incidence among men in New South Wales, Australia, 1987–2011

Objective：We assessed the trends in lung cancer incidence over a 25-year period by socioeconomic groups for men in New South Wales（NSW）,Australia.Methods：Men diagnosed with lung cancer between 1987 and 2011 were divided into five quintiles according to an Index of Education and Occupation（IEO）.We assessed relative socioeconomic differences over time by calculating age-standardized incidence ratios（SIRs）by 5-year period of diagnosis,and estimated absolute differences by comparing the observed and expected numbers of cases using the highest IEO quintile as the reference.Results：Lung cancer incidence for men decreased from 1987 to 2011 for all IEO quintiles,with a greater rate of decline for men living in the highest IEO areas.Thus,the relative disparity increased significantly over the 25-year period（P=0.0006）.For example,the SIR for the lowest IEO quintile increased from 1.28 during 1987–1991 to 1.74during 2007–2011.Absolute differences also increased with the proportion of＂potentially preventable＂cases doubling from 14.5% in 1987–1991 to 30.2% in 2007–2011.Conclusions：Despite the overall decline in lung cancer incidence among men in NSW over the past 25 years,there was a significant increase in disparity across socioeconomic areas in both relative and absolute terms.

昆虫性信息素的研究进展

药用昆虫作为中药的重要组成部分,相关医药研发日益获得重视。昆虫性信息素的研究应用也因此受到关注。目前,昆虫性信息素主要用于农林业害虫防治。本文在简述昆虫性信息素的化学成分、释放及应用的同时,对昆虫性信息素在昆虫大量诱捕、干扰交配、配合治虫等在药用昆虫资源的开发中的应用也进行初步探讨。

血必净注射液联合抗生素治疗脓毒症:两类药物间高水平药代和谐

联合用药既需要药物间药效协同互补,也需要药物间能够“药代和谐”(pharmacokinetic compatibility,PKC;不发生会影响药物有效性或安全性的药代性质药物相互作用)。当今世界天然产物制品与化药一同使用既大量存在,又充满争议。一方面,服用葡萄柚汁、圣约翰草制剂等能严重干扰同时进行的化药治疗,这使人们对天然产物制品与化药合用充满戒心;另一方面,越来越多严格的临床和基础研究证明,中药联合化药能更好地应对多因素疾病。脓毒症是一种由感染引起机体反应失调所导致危及生命的器官功能障碍,死亡率高、预后不良。

Microbial markers in colorectal cancer detection and/or prognosis

Colorectal cancer(CRC) is the second leading cause of cancer worldwide. CRC is still associated with a poor prognosis among patients with advanced disease. On the contrary, due to its slow progression from detectable precancerous lesions, the prognosis for patients with early stages of CRC is encouraging. While most robust methods are invasive and costly, actual patient-friendly screening methods for CRC suffer of lack of sensitivity and specificity. Therefore, the development of sensitive, non-invasive and cost-effective methods for CRC detection and prognosis are necessary for increasing the chances of a cure. Beyond its beneficial functions for the host, increasing evidence suggests that the intestinal microbiota is a key factor associated with carcinogenesis. Many clinical studies have reported a disruption in the gut microbiota balance and an alteration in the faecal metabolome of CRC patients, suggesting the potential use of a microbialbased test as a non-invasive diagnostic and/or prognostic tool for CRC screening. This review aims to discuss the microbial signatures associated with CRC known to date, including dysbiosis and faecal metabolome alterations, andthe potential use of microbial variation markers for noninvasive early diagnosis and/or prognostic assessment of CRC and advanced adenomas. We will finally discuss the possible use of these markers as predicators for treatment response and their limitations.

Enantiomeric characterization and structure elucidation of Otamixaban

Otamixaban is a potent （Ki=0.5 nM） fXa inhibitor currently in late-stage clinical develop-ment at Sanofi for the management of acute coronary syndrome. Being unproductive in obtaining a suitable crystal of Otamixaban, the required enantiomeric characterization has been accomplished using vibrational circular dichroism （VCD） spectroscopy. Selected by a spectrum similarity index, the calculated spectra of several higher energy conformers were found to match well with the observed spectra. The characteristic IR bands of these conformers were also identified and attributed to the solvation effect. Combined with both the single crystal x-ray diffraction results for an intermediate and the proton NMR study, the absolute configuration of Otamixaban is unambiguously determined to be （R,R）.

Long-term safety follow-up of children from a randomized-controlled phase Ⅱb proof-of-concept efficacy study of the live,attenuated,tetravalent dengue vaccine (CYD-TDV) in Thailand

Objective:To investigate the long-term safety of a tetravalent dengue vaccine(CYD-TDV)in children in a phase Ⅱ b follow-up study in Thailand.Methods:In the phase Ⅱb study,children aged 4-11 years were randomized(2:1)to receive three injections of CYD-TDV or serve as control at 6-month intervals,with 25 months’ active follow-up(active phase).This study was an additional four-year passive surveillance for hospitalized virologically-confirmed dengue(VCD;hospital phase).Cases of hospitalized VCD,severe hospitalized VCD,vaccine-related serious adverse events,and deaths were reported for the total population,with post-hoc analyses by enrollment age(<9 and≥9 years).Results:Of 3 997 participants receiving≥1 injection,80.1% were recruited to the hospital phase[2 131(CYD-TDV);1 072(control)].Eighty-five hospitalized VCD cases were reported in the CYD-TDV group and 46 in the control group during the four-year hospital phase[relative risk(RR):0.93,95% confidence interval(Cl):0.64-1.36].The RR over six years of follow-up was 0.77(95% CI:0.57-1.05).In those aged≥9 years,the cumulative RRs in the active phase,hospital phase,and entire six years were 0.28(95% CI:0.08-0.81),0.51(95% CI:0.25-1.05),and 0.42(95% CI:0.24-0.75),respectively.In the overall population,there were ten severe hospitalized VCD cases in the CYD-TDV group and five in the control group over six years(RR:1.00,95% CI:0.31-3.75).Conclusions:Over six years of follow-up,in children aged ≥9 years,CYD-TDV administration is associated with a reduced risk of hospitalized VCD.

静脉注射给药的甘草酸易在由肝转运体OATP1B1/OATP1B3介导的药物相互作用中成为被影响药物

随意使用天然产品可能严重干扰患者正在接受的化药治疗,此已引起国际广泛关注。中西医结合治疗疾病在中国很普遍,研究中药与化药合用时发生药代性质药物相互作用风险,对于保证临床用药的有效性和安全性十分重要。不同于美英等西方国家,中国将中药作为药物来监管和使用,因此研究中药与化药合用的风险既涉及“中药影响化药”,也涉及“化药影响中药”。国际上围绕化药影响天然产物制品的研究还很少,这是因为与健康相关的天然产物制品在西方国家通常不具备药物身份,缺乏相应的药代动力学研究和技术。研究“化药影响中药”的风险首先应明确成分体内暴露改变能够影响中药的有效性或安全性,其重点是要搞清楚与化药合用能否通过某种机制改变这些中药成分的体内暴露。

Clinical development update: 1st dengue vaccine candidate

A candidate tetravalent dengue vaccine is being assessed in three clinical trials involving more than 35,000 children between the ages of 2 and 16 years in the Asia-Pacific and Latin American countries.We report the results of long-term follow-up interim analyses and integrated efficacy analyses.We are assessing the incidence of hospitalization for virologically confirmed dengues a surrogate safety end point during follow-up in years 3–6 of two phase 3 trials,CYD14 and CYD15,and a phase 2b trial,CYD23/57.We estimated vaccine efficacy using pooled data from the first 25 months of CYD14 and CYD15.

Glargine-300: An updated literature review on randomized controlled trials and real-world studies

Despite the availability of a variety of insulins,rates of insulinisation and the acceptance of insulin therapy is suboptimal in real-world clinical settings.Patient and physician concerns with hypoglycaemia and weight gain are the two key issues that serve to impede appropriate insulinisation in patients with diabetes.Recently introduced second-generation basal insulin analogues [for e.g.,insulin glargine 300 U/m L(Gla-300) and insulin degludec] are designed to have improved pharmacokinetic profiles with an intention to deliver steady insulin levels over a longer period.Several randomised controlled and real-world studies have proven the resultant advantages of second-generations insulin analogues in lowering intra-individual variability in plasma insulin levels,flexibility in dosing,a sustained glucose-lowering effect,and decreasing the risk of hypoglycaemia.Gla-300 is one of the newer second-generation basal insulin analogues to have been approved for both type 1 and 2 diabetes.In this article,we review the currently available clinical and real-world data of Gla-300.

Release of exosomes and microvesicles harbouring specific RNAs and glycosylphosphatidylinositol-anchored proteins from rat and human adipocytes is controlled by histone methylation

The transfer of proteins and nucleic acids from donor to acceptor cells via small membrane vesicles has been implicated with (patho)physiological consequences. Previously the upregulation of esterification and downregulation of lipolysis in small rat adipocytes upon incubation with exosomes and microvesicles (EMVs) released from large adipocytes and harbouring the glycosylphosphatidylinositol (GPI)-anchored proteins, Gce1 and CD73, transcripts specific for FSP27 and GPAT3, and microRNAs, miR-16 and miR-222 was demonstrated. Here the release of EMVs from large (but not small) primary and differentiated and human rat adipocytes in response to palmitate, H2O2 and the anti-diabetic sulfonylurea, glimepiride, is shown to be significantly reduced upon inhibition of histone H3 lysine9 methyltransferase G9a by trans-2-phenylcyclopropylamine (tPCPA) and histone H3 lysine4 demethylase LSD1 by BIX01294. Inhibition of EMV release by tPCPA and BIX01294 was not caused by apoptosis but accompanied by upregulation of the H2O2-induced stimulation of lipid synthesis and downregulation of lipolysis in large (but not small) primary and differentiated rat and human adipocytes. In contrast, the simultaneous presence of tPCPA and BIX-01294 had almost no effect on the induced release of EMVs and lipid metabolism. These findings argue for regulation of the release of EMVs harbouring specific GPI-anchored proteins, transcripts and microRNAs from rat and human adipocytes by histone H3 methylation at lysines 4 and 9 in interdependent fashion. Thus the EMV-mediated transfer of lipogenic and anti-lipolytic information between large and small adipocytes in response to certain physiological and pharmacological stimuli seems to be controlled by epigenetic mechanisms.

Effect of Initiation of Basal Insulin Glargine on Glycemic Control in Patients with Diabetes: Real Life Experience from Hong Kong

Introduction: To assess the changes in glycemic control after initiating or switching to a basal insulin analogue in patients with diabetes mellitus. Methods: A retrospective, observational analysis was conducted using electronic data from a Hong Kong regional hospital. Data from adult patients with type 1 and 2 diabetes mellitus (T1DM and T2DM, respectively) who had been prescribed with basal insulin glargine in 2008-2010, with recorded HbA1c levels at the time of initiation, at 6 and 12 months thereafter, were analysed. Results: Data from 106 eligible patients were analysed. Substantial reduction in HbA1c and fasting sugar levels were reported in both T1DM (Δ HbA1c = 1.5%, Δ FBG = 1.3 mmol/L p < 0.05) and T2DM (Δ HbA1c = 1.2%, Δ FBG = 2.9 mmol/L p < 0.05) patients after 12 months of therapy. A total of 42% of T1DM and 26% of T2DM patients achieved HbA1c levels < 7.0%. After adjustment, T2DM patients who were insulin naive achieved a statistically greater HbA1c reduction (Δ = 1.7%) than those who previous treated with premixed or basal bolus insulin (Δ = 0.3%) (p < 0.05). Percentage of patients experiencing hypoglycaemia reduced from 69% to 62% in T1DM but increased from 26% to 36% in T2DM patients. All hypoglycaemic episodes recorded were either asymptomatic or mild and self-limiting. Only 4% of the patients discontinued treatment at the end of 12 months. Conclusions: In real life clinical practice, a single daily basal insulin analogue therapy provided effective glycemic control with an acceptable risk of mild hypoglycaemia.

Concordance Between Laboratories Testing Clinical Samples and Its Impact on Vaccine Booster Thresholds

Concordance study between two laboratories testing clinical samples is important to a clinical program to assess the comparability of test results, especially when the two laboratories performed the same test at different locations. Assays that measure immune response should not be compared using the simple linear regression model; instead literature publications recommend an ＂errors-in-variables＂ model that accounts for the variability in these assays （Home, 1998）. In this paper, concordance slope and intercept was estimated using the statistical linear relationship （Tan and Iglewicz, 1999） for various tests between laboratories. The confidence bounds for the concordance line were used to assess the impact on vaccine booster thresholds when a consistent bias was observed between the results generated by two laboratories.

Change-Point Analysis of Survival Data with Application in Clinical Trials

Effects of many medical procedures appear after a time lag, when a significant change occurs in subjects’ failure rate. This paper focuses on the detection and estimation of such changes which is important for the evaluation and comparison of treatments and prediction of their effects. Unlike the classical change-point model, measurements may still be identically distributed, and the change point is a parameter of their common survival function. Some of the classical change-point detection techniques can still be used but the results are different. Contrary to the classical model, the maximum likelihood estimator of a change point appears consistent, even in presence of nuisance parameters. However, a more efficient procedure can be derived from Kaplan-Meier estimation of the survival function followed by the least-squares estimation of the change point. Strong consistency of these estimation schemes is proved. The finite-sample properties are examined by a Monte Carlo study. Proposed methods are applied to a recent clinical trial of the treatment program for strong drug dependence.

Antimicrobial Activity of a Cys-Rich Peptide Derived from a Centrosema virginianum Vicilin

Many antimicrobial peptides (AMPs) have been identified in plants. These peptides are highly divergent at the primary sequence level and vary in their hierarchical structures. Some common biochemical features include the ability to form disulfide bonds, tandemly repeated amino acid sequences and a net charge at pH 7. Unusual Cysteine containing repeats has been identified in several plant seed storage proteins that may act as AMPs. We identified a Cys repeat within a vicilin (seed storage protein) of a wild legume, Centrosema virginianum. Cleavage of the vicilin protein during germination would generate a vicilin derived Cys peptide (VDCP). We investigated the antimicrobial properties of this VDCP and compared its efficacy as an antimicrobial agent to VDCPs from other species. We developed transgenic tobacco plants that expressed cloned sequences encoding the Cysteine repeat unit from C. virginianum, Theobroma cacao and Gossypium hirsutum. Extracts from fully expanded leaves were tested for antimicrobial activity against a fungal pathogen, Botrytis cinerea. The Cys motif from C. virginianum was also expressed in two E. coli cell lines (reducing or oxidizing cytoplasm) and peptide fusion protein fractions were tested for antimicrobial activity against a battery of fungal strains. The unique Cysteine repeat single unit from C. virginianum exhibited antimicrobial properties greater than or equal to the antimicrobial activity associated with expression of the multiple Cys-repeat VDCPs from G. hirsutum or T. cacao in transgenic tobacco. When expressed in bacteria, a C. virginianum VDCP fusion protein exhibited antifungal activity against 3 of the 4 fungi tested. Although the primary role of seed storage proteins is to provide a pool of amino acids and nitrogen for germinating seeds and developing plantlets, it is likely that seed storage protein proteolytic products also provide beneficial antimicrobial properties during germination and young plantlet development.

Insulin Glargine 300 Units/mL Effectiveness in Patients with T2DM Uncontrolled by Basal Insulin in Real-Life Settings in the Czech Republic

Introduction: To evaluate the clinical effectiveness of Gla-300 units/mL (Gla-300) in the treatment of patients with type 2 diabetes (T2DM) uncontrolled by basal insulin in real-life clinical settings in the Czech Republic (TOPAZ study). Methods: TOPAZ was a prospective, multi-center, non-interventional, 6-month study. Of the 312 patients screened, 289 were evaluated at month 6. The primary objective was the change of HbA1c after 6 months. The proportion of patients with HbA1c < 7.0% DCCT (< 53 mmol/mol), and those with a decrease of at least 0.5% of HbA1c at month 6, change in FPG, body weight and insulin dose at month 3 and 6 were analysed as secondary objectives. Incidence of hypoglycemia, adverse events and patient treatment satisfaction were also assessed. Results: HbA1c decreased significantly after 6 months (mean change 0.9% ± 1.1% DCCT [−9.9 ± 11.6 mmol/mol], p < 0.0001). HbA1c target < 7.0% DCCT was achieved in 17.6% of patients, 66.1% of patients showed mean HbA1c decrease of 0.5% ± 0.8%. At month 6, FPG decreased (mean change from baseline −1.8 ± 3.1 mmol/L) as well as the incidence of hypoglycemia decreased by 49% (p < 0.0001) while no weight gain was observed. No significant safety signals were identified. Conclusion: In a real-life setting, switching to Gla-300 in T2DM patients uncontrolled with other basal insulin was associated with improved glycemic control and reduced risk of hypoglycemia without weight gain, while patients’ satisfaction with treatment increased.