Cohort study of HIV-positive and -negative methamphetamine users

Background

The effects of methamphetamine (MA) on caries have been well documented. Little, however, is known about its effects on the periodontium. The authors conducted this study to determine the prevalence and severity of periodontal disease in an urban population of HIV-positive MA users.

Evaluation of Pre-analytical Variables in a Commercial Thrombin Generation Assay

Background

There is minimal data on the influence of pre-analytical variables on the use of calibrated automated thrombography (CAT), to measure thrombin generation.

Objectives

To evaluate the impact of centrifugation methods, time after collection, and contact activation inhibition on the CAT assay performed using two commercial reagents.

Methods and Results

Six different methods of plasma separation were examined. Thrombin generation triggered by a 5 pM tissue factor reagent was not greatly affected by plasma separation method, with similar results obtained with all methods apart from single centrifugation and membrane filtration. Membrane filtration increased APTT and is not recommended. Extended double centrifugation at higher speed was required to minimise the impact of residual phospholipid with 1 pM tissue factor trigger, particularly with inhibition of contact activation. The effect of a delay of up to 24 hours in preparing plasma was assessed. No significant difference in results was observed among samples processed between 0.5 and 6 hours after blood collection into plastic Vacuette® tubes. The presence or absence of corn trypsin inhibitor had a significant impact on all parameters with 1 pM tissue factor trigger, with minor differences seen on Peak and ttPeak results using 5 pM tissue factor.

Conclusions

The impact of pre-analytical variables on thrombin generation results is dependent on the concentration of tissue factor in the trigger reagent used. Results with 1 pM tissue factor are particularly sensitive to centrifugation method and contact activation, and standardisation is required to allow large collaborative studies to be performed.     

Liver Renewal: Detecting Misrepair and Optimizing Regeneration

Cirrhosis and liver cancer, the main causes of liver-related morbidity and mortality, result from defective repair of liver injury. This article summarizes rapidly evolving knowledge about liver myofibroblasts and progenitors, the 2 key cell types that interact to orchestrate effective repair, because deregulation of these cells is likely to be central to the pathogenesis of both cirrhosis and liver cancer. We focus on cirrhosis pathogenesis because cirrhosis is the main risk factor for primary liver cancer. Emerging evidence suggests that the defective repair process has certain characteristics that might be exploited for biomarker development. Recent findings in preclinical models also indicate that the newly identified cellular and molecular targets are amenable to therapeutic manipulation. Thus, recent advances in our understanding about key cell types and fundamental mechanisms that regulate liver regeneration have opened new avenues to improve the outcomes of liver injury.Trial Registrationclinicaltrials.gov Identifier: NCT01899859     

The mechanism of action of MF59 - an innately attractive adjuvant formulation

MF59 is a safe and effective vaccine adjuvant which was originally approved to be included in a licensed influenza vaccine to be used in the elderly in Europe in 1997. The MF59 adjuvanted influenza vaccine (Fluad™) is now licensed in more than 20 countries worldwide and more than 85 million doses have been administered. More recently the vaccine adjuvant has also been shown to be safe and effective in young children and resulted in a significant increase in influenza vaccine efficacy in a controlled clinical trial in Europe. Since the early days of its discovery we have explored the mechanism of action of MF59, using a variety of available techniques. In recent years we have explored more thoroughly the mechanism of action using new and more sophisticated techniques. It is remarkable how consistent the data has been, using a variety of different approaches both in several small animal models and also using human immune cells in vitro. Here we present a summary of all the work performed to date on the mechanism of action of MF59 and we present a unified theory based on the accumulated data of how it exerts its adjuvant effects. A key element of the mechanism of action appears to be the creation of a transient ‘immunocompetent’ local environment at the injection site, resulting in the recruitment of key immune cells, which are able to take up antigen and adjuvant and transport them to the local lymph nodes, where the immune response is induced. This recruitment appears to be triggered by the induction of a chemokine driven gradient by the impact of MF59 on local cells, which are activated to secrete further chemokines, which are recruitment factors for more immune cells.     

蕈样肉芽肿皮损及外周血人类疱疹病毒1、2、4、5型DNA的检测

目的　探讨蕈样肉芽肿与人类疱疹病毒 1型 (单纯疱疹病毒 1型 )、2型 (单纯疱疹病毒 2型 )、4型 (ep stein barr病毒 )、5型 (人类巨细胞病毒 )间有无相关性。方法　酚 氯仿法提取皮肤组织 (蕈样肉芽肿 3 2份、正常皮肤 2 9份、慢性湿疹与慢性单纯性苔藓 2 4份 )及外周血白细胞 (蕈样肉芽肿 15份、健康献血员 40份、慢性湿疹与慢性单纯性苔藓 11份 )中基因组DNA。采用聚合酶链反应技术检测病毒DNA ,扩增产物回收后以限制性内切酶BamHⅠ、SmaⅠ酶切鉴定。结果　蕈样肉芽肿皮损及外周血中 ,各有 2份检测到人类疱疹病毒 1型DNA ;蕈样肉芽肿皮损中有 5份存在人类疱疹病毒 4型DNA ;健康献血员外周血以及正常皮肤、慢性湿疹与慢性单纯性苔藓患者皮损组织及外周血中未检测到此4种病毒DNA。结论　蕈样肉芽肿与人类疱疹病毒 1、2、4、5型间可能不具有相关性。     

Hypermethylated DAPK in serum DNA of women with uterine leiomyoma is a biomarker not restricted to cancer

Objective

Ovarian cancer is most frequently diagnosed at a late stage with a poor prognosis. No markers for early diagnosis have been established. Aberrantly methylated DNA appears as a promising molecular cancer marker. The aim of this study was to analyze the methylation status of the proapoptotic cancer related gene death-associated protein kinase (DAPK) in ovarian cancer patients, healthy controls and in patients suffering from a benign proliferative disease such as uterine leiomyoma.

Methods

Methylation-specific PCR (MSP) was used to detect DAPK methylation in primary tumor tissue and serum of both ovarian cancer (n = 32) and uterine leiomyoma patients (n = 17 primary tissue, n = 30 serum). Serum samples from healthy women served as controls (n = 20). MSP results were confirmed by restriction digest and sequencing analyses of cloned PCR products.

Results

DAPK methylation was detected in 50% and 35.3% of primary tissue and 56% and 23.8% of serum samples from ovarian cancer and leiomyoma patients, respectively. However, the association of methylation frequencies in tissue and serum was low (kappa = −0.053). Sequencing experiments revealed fully methylated MSP products in sera of both ovarian cancer and leiomyoma patients. In contrast sera from control patients showed only partially methylated DAPK sequences.

Conclusion

DAPK hypermethylation was neither specific for the tissue of origin nor for cancer. The high prevalence of leiomyoma compromises the utility of this gene as a serum marker for early ovarian cancer detection. These data emphasize the necessity to co-analyze controls presenting with non-cancer proliferative disease in the quest for molecular cancer markers.     

MF59 Emulsion Is an Effective Delivery System for a Synthetic TLR4 Agonist (E6020)

Purpose  The effectiveness of vaccines depends on the age and immunocompetence of the vaccinee. Conventional non-adjuvanted influenza vaccines are suboptimal in the elderly and vaccines with improved ability to prevent influenza are required. The TLR4 agonist E6020, either given alone or co-delivered with MF59, was evaluated and compared to MF59 and the TLR9 agonist CpG. Its ability to enhance antibody titres and to modulate the quality of the immune response to a subunit influenza vaccine was investigated. Methods  Mice were immunized with either antigens alone, with MF59 or with the TLR agonists alone, or with a combination thereof. Serum samples were assayed for IgG antibody titres and hemagglutination inhibition (HI) titres. Th1/Th2 type responses were determined by titrating IgG subclasses in serum samples and by T-cell cytokine responses in splenocytes. Results  MF59 was the best single adjuvant inducing HI and T-cell responses in comparison to all alternatives. The co-delivery of E6020 or CpG with MF59 did not further increase antibody titres however shifted towards a more Th1 based immune response. Conclusion  Combining adjuvants like E6020 and MF59 allowed a finer tuning of the immune response towards a particular Th bias, thus have significant implications for the development of improved influenza vaccines.     

Sézary syndrome without erythroderma: A case report and review of published work

Sézary syndrome (SS) is defined by erythroderma and circulating atypical T cells, with or without lymphadenopathy. Recently, Thompson et al. identified a distinct population of SS patients with an atypical presentation: a high blood tumor burden of Sézary cells fulfilling criteria for SS but without fulfilling the criteria for erythroderma at the diagnosis. Here, we report a case of a 49‐year‐old Japanese man with SS who did not present with erythroderma initially, but exhibited erythematous itchy papules symmetrically located on the legs and arms. We also reviewed reported cases of SS without initial erythroderma. The skin manifestations at diagnosis varied from patches to tumors often seen in mycosis fungoides, and other rarer findings such as excoriation, palmoplantar keratoderma and alopecia. Pruritus was reported in most patients (86%), unlike early mycosis fungoides, and could be the main clue to the diagnosis of SS. Notably, three patients were reported to have presented with papular lesions, similar to our case. Little is known about why skin lesions in SS without erythroderma vary and why these cases did not exhibit erythroderma initially. Attenuated stimulation by colonized Staphylococcus aureus, impairment in recruitment of malignant T cells and suppression of inflammatory response induced by malignant T cells with regulatory phenotype may be associated with skin manifestations. Further studies are necessary to elucidate the etiology of this entity.     

Higher prevalence of thyroid dysfunction in patients with erythrodermic psoriasis

The association of psoriasis with thyroid dysfunction has been investigated. However, it remains unclear; some papers indicate it, and others do not. In this study, we evaluate the prevalence of thyroid dysfunction in patients with psoriasis vulgaris (PsV), psoriatic arthritis (PsA), generalized pustular psoriasis (GPP) and erythrodermic psoriasis (EP), and the association of thyroid dysfunction with inflammation. Data on 201 psoriatic patients visiting our hospital from January 2014 to November 2017 (159 men and 42 women; 74 PsV, 42 PsA, 38 GPP and 47 EP) were retrospectively analyzed. Thirty-three percent of psoriatic patients had thyroid dysfunction. The percentage of patients with thyroid dysfunction was the highest in those with EP (60% EP, 42% GPP, 19% PsA, 19% PsV). The prevalence of thyroid dysfunction decreased significantly when patients switched from EP to PsV or PsA (58% vs 17%; median, 20.5; range, 4–65 months). Most of the patients with thyroid dysfunction had low thyroxine syndrome (serum levels of free thyroxine are low, but serum thyroid-stimulating hormone level is normal). Patients with thyroid dysfunction demonstrated significantly higher CD3⁺ and CD4⁺ T-cell absolute count levels than those without thyroid dysfunction. Meanwhile, patients with thyroid dysfunction demonstrated lower immunoglobulin (Ig)A and IgM levels than those without thyroid dysfunction. Finally, patients with thyroid dysfunction demonstrated higher elevated serum C-reactive protein levels than those without dysfunction in total, although there were no statistical differences. Our data indicate that thyroid dysfunction in patients with psoriasis may be associated with inflammation caused by psoriasis.