Pathways to HIV risk and vulnerability among lesbian,gay, bisexual,and transgendered methamphetamine users: a multi-cohort gender-based analysis

Background  

Methamphetamine (MA) use continues to be a major public health concern in many urban settings. We sought to assess potential relationships between MA use and individual, social, and structural HIV vulnerabilities among sexual minority (lesbian, gay, bisexual or transgendered) drug users.     

Toll-Like Receptor 4 Signaling Leads to Severe Fungal Infection Associated with Enhanced Proinflammatory Immunity and Impaired Expansion of Regulatory T Cells

Toll-like receptors (TLRs) present in innate immune cells recognize pathogen molecular patterns and influence immunity to control the host-parasite interaction. The objective of this study was to characterize the involvement of TLR4 in the innate and adaptive immunity to Paracoccidioides brasiliensis, the most important primary fungal pathogen of Latin America. We compared the responses of C3H/HeJ mice, which are naturally defective in TLR4 signaling, with those of C3H/HePas mice, which express functional receptors, after in vitro and in vivo infection with P. brasiliensis. Unexpectedly, we verified that TLR4-defective macrophages infected in vitro with P. brasiliensis presented decreased fungal loads associated with impaired synthesis of nitric oxide, interleukin-12 (IL-12), and macrophage chemotactic protein 1 (MCP-1). After intratracheal infection with 1 million yeasts, TLR4-defective mice developed reduced fungal burdens and decreased levels of pulmonary nitric oxide, proinflammatory cytokines, and antibodies. TLR4-competent mice produced elevated levels of IL-12 and tumor necrosis factor alpha (TNF-α), besides cytokines of the Th17 pattern, indicating a proinflammatory role for TLR4 signaling. The more severe infection of TLR4-normal mice resulted in increased influx of activated macrophages and T cells to the lungs and progressive control of fungal burdens but impaired expansion of regulatory T cells (Treg cells). In contrast, TLR4-defective mice were not able to clear their diminished fungal burdens totally, a defect associated with deficient activation of T-cell immunity and enhanced development of Treg cells. These divergent patterns of immunity, however, resulted in equivalent mortality rates, indicating that control of elevated fungal growth mediated by vigorous inflammatory reactions is as deleterious to the hosts as low fungal loads inefficiently controlled by limited inflammatory reactions.Pathogen recognition receptors (PRRs) are a group of receptors present in the membrane and cytoplasm of innate immunity cells that recognize the presence of invading microbes by interacting with conserved pathogen structures, the so called “pathogen-associated molecular patterns” (PAMPs). This initial event of innate immunity is crucial for the control of pathogen growth and the subsequent activation of adaptive immunity. Toll like receptors (TLRs) constitute a major family of pattern recognition molecules and, like other PRRs, are able to respond to different structural homologies conserved in many microorganisms (2, 62). Activation of the TLRs is crucial for many aspects of microbe elimination, including microbial killing, recruitment of phagocytes to the site of infection, and activation of dendritic cells (DCs) (52). Early TLR activation results in the production of several inflammatory mediators, and the final balance among pro- and anti-inflammatory components regulates the type of adaptive immune response. Recent findings have shown that direct recognition of PAMPs by DCs is critical for priming appropriate T-cell responses, resulting in T helper 1 (Th1), Th2, or Th17 immunity (25, 31, 33, 60).TLR4 is the key receptor that recognizes bacterial lipopolysaccharides (LPS), whereas TLR2 is involved in the interaction with bacterial peptidoglycans and lipoproteins (66). As reported for other microorganisms, TLRs have been shown to be involved in host defense against different fungal pathogens. In vivo and in vitro studies have demonstrated that Cryptococcus neoformans (7, 67), Candida albicans (43, 45), and Aspergillus fumigatus (24, 41) may signal through members of the TLR family, mainly TLR2 and TLR4. Different components of a certain pathogen can be used to stimulate the immune system. Thus, C. albicans phospholipomannan is sensed by TLR2 (34), while O-linked mannans are recognized by TLR4 (44). The contribution of individual TLRs to the immune response against pathogenic fungi depends on several factors, such as the fungal morphotype, fungal species, and route of infection. Activation signals mediated by innate immunity receptors, however, are not always beneficial to the host, and TLR activation can be used by pathogenic fungi to promote more-severe infections (6, 53).Paracoccidioidomycosis (PCM) is a systemic granulomatous disease caused by the dimorphic fungus Paracoccidioides brasiliensis and constitutes the most prevalent deep mycosis in Latin America (28). The alveolar macrophages are the first host cells that interact with P. brasiliensis cells, and their activation is fundamental to the control of fungal growth. The molecular mechanisms controlling the initial steps of the interaction between P. brasiliensis and phagocytes are not well understood. It is known, however, that normal macrophages are permissive to P. brasiliensis growth, while cytokine-activated macrophages are able to restrain P. brasiliensis multiplication (12). Complement receptor 3 (CR3) and mannose receptor have been shown to play important roles in the initial interaction between P. brasiliensis cells and mouse peritoneal macrophages (14, 32, 50). Interestingly, recent work from our laboratory demonstrated that alveolar macrophages from susceptible mice (B10.A) are easily activated by P. brasiliensis infection and show efficient fungal killing associated with nitric oxide production, while pulmonary macrophages from resistant mice (A/Sn) are poorly activated and present inefficient killing activity associated with increased levels of transforming growth factor β (TGF-β) (49). Despite their inefficient innate immunity, A/Sn mice develop a balanced Th1/Th2 immune response that controls fungal growth without intense tissue pathology.In previous work, our group demonstrated the influence of TLR2 on pulmonary PCM (15, 38). Using TLR2-normal and TLR2-deficient mice, we were able to show that the presence of TLR2 increases the severity of in vitro and in vivo P. brasiliensis infections. In addition, TLR2 deficiency results in increased Th17 immunity associated with diminished expansion of regulatory T cells (Treg cells) and increased lung pathology due to unrestrained inflammatory reactions (38). Characterizing the behavior of dendritic cells in murine PCM, Ferreira et al. observed increased expression of TLR2 by dendritic cells of susceptible, but not resistant, mice (26). Moreover, it has been suggested that TLR2, TLR-4, and dectin-1 are involved in the recognition and internalization of P. brasiliensis by human monocytes and neutrophils, indicating important roles for these pathogen receptors in the immune response against the fungus (10).We decided to investigate the role of TLR4 in murine PCM by using in vitro and in vivo models of infection. Using TLR-competent C3H/HePas mice and C3H/HeJ mice, which possess a missense mutation in the TLR4 gene, we were able to demonstrate that both in vitro and in vivo, TLR4 signaling increases the severity of infection in association with increased activation of innate and adaptive immunity but decreased expansion of Treg cells. This pattern of immunity, however, was not beneficial to the hosts, due to the increased lung injury mediated by inefficient control of inflammatory reactions by Treg cells.     

CD28 Exerts Protective and Detrimental Effects in a Pulmonary Model of Paracoccidioidomycosis

T-cell immunity has been claimed as the main immunoprotective mechanism against Paracoccidioides brasiliensis infection, the most important fungal infection in Latin America. As the initial events that control T-cell activation in paracoccidioidomycosis (PCM) are not well established, we decided to investigate the role of CD28, an important costimulatory molecule for the activation of effector and regulatory T cells, in the immunity against this pulmonary pathogen. Using CD28-deficient (CD28^−/−) and normal wild-type (WT) C57BL/6 mice, we were able to demonstrate that CD28 costimulation determines in pulmonary paracoccidioidomycosis an early immunoprotection but a late deleterious effect associated with impaired immunity and uncontrolled fungal growth. Up to week 10 postinfection, CD28^−/− mice presented increased pulmonary and hepatic fungal loads allied with diminished production of antibodies and pro- and anti-inflammatory cytokines besides impaired activation and migration of effector and regulatory T (Treg) cells to the lungs. Unexpectedly, CD28-sufficient mice progressively lost the control of fungal growth, resulting in an increased mortality associated with persistent presence of Treg cells, deactivation of inflammatory macrophages and T cells, prevalent presence of anti-inflammatory cytokines, elevated fungal burdens, and extensive hepatic lesions. As a whole, our findings suggest that CD28 is required for the early protective T-cell responses to P. brasiliensis infection, but it also induces the expansion of regulatory circuits that lately impair adaptive immunity, allowing uncontrolled fungal growth and overwhelming infection, which leads to precocious mortality of mice.It has long been appreciated that cellular immunity is the most important resistance mechanism against fungal infections (14, 36, 64). CD4⁺ and CD8⁺ T-cell subpopulations have been described to have a fundamental role in the control of fungal growth, and disease severity is also controlled by regulatory T (Treg) cells, which prevent tissue pathology by controlling excessive inflammatory reactions (25, 45, 46, 65). Similar to other deep mycoses, the severity of paracoccidioidomycosis (PCM), the most severe pulmonary mycosis in Latin America, is controlled by cellular immunity and cytokine-activated phagocytes that are able to kill Paracoccidioides brasiliensis, the etiological agent of this infection (10, 20, 30, 60, 61). In humans and in murine models of PCM, resistance to the disease is associated with the secretion of gamma interferon (IFN-γ) and other type 1 cytokines, whereas impaired Th1 immunity and the prevalent secretion of Th2 cytokines correlate with a systemic and progressive disease (2, 6, 39, 59, 76). Studies with CD4⁺ and CD8⁺ T-cell-deficient mice revealed that both T-cell subsets are involved in the protective immunity against P. brasiliensis infection and indicated the prominent role of CD8⁺ T cells (3, 21, 25). Besides the prevalent Th2 immunity, recent investigations have described alternative mechanisms underlying T-cell dysfunction in humans and experimental PCM. Increased apoptosis and overexpression of Fas and FasL in T cells suggest that activation-induced cell death (AICD) is a mechanism that controls T-cell expansion during the active disease (13, 19). In addition, the increased expression of CTLA-4 and the expansion of Treg cells were associated with severe patterns of the disease (24, 45, 46, 56). Thus, in addition to cytokine imbalance, other regulatory mechanisms can actively participate in the unresponsiveness of T cells in P. brasiliensis-infected hosts.Optimal activation, proliferation, and cytokine production by antigen-specific T cells require two distinct signals from dendritic cells or other antigen-presenting cells (APCs). After T-cell receptor (TCR) occupancy by the antigen epitope/major histocompatibility (MHC) complex (first signal), a second signal is mediated by costimulatory molecules (43, 63), such as CD28 on T cells and their counter-receptors CD80 (B7-1) and CD86 (B7-2) expressed by APCs (1, 34). Soluble molecules, such as cytokines and chemokines, also participate in the activation process, which drives and controls T-cell numbers and fates (1). CD28 enhances the TCR-triggered activation of naïve T cells, promotes interleukin-2 (IL-2) secretion and prevents T-cell anergy (1, 37). Alternatively, CD28-independent T-cell activation can occur if a strong and sustained antigen-specific signal is available (40, 81). Like CD28, two other molecules, cytotoxic T-lymphocyte antigen-4 (CTLA-4) and mouse inducible costimulatory molecule (ICOS), are selectively expressed by T cells, but the expression of these molecules depends on previous cell activation (50, 71). More recently, evidence has emerged that CD28 family members are also crucial regulators of natural and induced regulatory (CD4⁺CD25⁺Foxp3⁺) T cells (9). These cells are induced in the thymus and in the periphery, respectively, and control self-tolerance and the activation of several components of innate and adaptive immunity (68). Treg cells can suppress immune responses through the production of immunosuppressive cytokines (mainly IL-10 and transforming growth factor β [TGF-β]), through the induction of the apoptosis of effector T cells and through the modification of the functional properties of antigen-presenting cells (70, 78).Immunoprotection against microorganisms has been shown to be either CD28 dependent or independent. CD28-deficient (CD28^−/−) mice are highly susceptible to infection with Salmonella enterica serovar Typhimurium due to the poor ability of these mice to secrete IFN-γ (51). During some viral and parasitic infections, CD28 was shown to be required to mediate CD8⁺ T-cell immunoprotection (8, 53). In contrast, CD28^−/− mice infected with Mycobacterium bovis or Listeria monocytogenes control the bacterial burden and develop cell-mediated immunity (35, 52). In primary and opportunistic fungal infections, CD28 costimulation controls protective immunity, the expansion and function of regulatory T cells, and the intensity of inflammatory reactions (5, 54, 55, 66, 84).Because CD28 is critical for T-cell activation in fungal infections, we investigated its role in a murine model of P. brasiliensis infection. We show that CD28 costimulation exerts contrasting roles in pulmonary PCM. Early in infection, CD28 expression results in efficient adaptive immunity that is able to control fungal growth. Late in infection, however, this costimulatory molecule induces significant expansion of regulatory T cells, diminished immunity, and uncontrolled fungal growth that eventually leads to the death of the mice. In contrast, the absence of CD28 costimulation results in impaired T-cell immunity, which appeared to be compensated by the absence of Treg cell expansion. This weak but persistent immunity was able to partially control fungal growth, organize granulomatous lesions, and guarantee the enhanced survival of the mice, suggesting the relative protection conferred by CD28-independent mechanisms.     

Preclinical characterization of WAY-211612: a dual 5-HT uptake inhibitor and 5-HT1A receptor antagonist and potential novel antidepressant

Background and purpose

As a combination of 5-HT selective reuptake inhibitor (SSRI) with 5-HT_1A receptor antagonism may yield a rapidly acting antidepressant, WAY-211612, a compound with both SSRI and 5-HT_1A receptor antagonist activities, was evaluated in preclinical models.

Experimental approach

Occupancy studies confirmed the mechanism of action of WAY-211612, while its in vivo profile was characterized in microdialysis and behavioural models.

Key results

WAY-211612 inhibited 5-HT reuptake (K_i = 1.5 nmol·L⁻¹; K_B = 17.7 nmol·L⁻¹) and exhibited full 5-HT_1A receptor antagonist activity (K_i = 1.2 nmol·L⁻¹; K_B = 6.3 nmol·L⁻¹; I_max 100% in adenyl cyclase assays; K_B = 19.8 nmol·L⁻¹; I_max 100% in GTPγS). WAY-211612 (3 and 30 mg·kg⁻¹, po) occupied 5-HT reuptake sites in rat prefrontal cortex (56.6% and 73.6% respectively) and hippocampus (52.2% and 78.5%), and 5-HT_1A receptors in the prefrontal cortex (6.7% and 44.7%), hippocampus (8.3% and 48.6%) and dorsal raphe (15% and 83%). Acute or chronic treatment with WAY-211612 (3–30 mg·kg⁻¹, po) raised levels of cortical 5-HT approximately twofold, as also observed with a combination of an SSRI (fluoxetine; 30 mg·kg⁻¹, s.c.) and a 5-HT_1A antagonist (WAY-100635; 0.3 mg·kg⁻¹, s.c). WAY-211612 (3.3–30 mg·kg⁻¹, s.c.) decreased aggressive behaviour in the resident-intruder model, while increasing the number of punished crossings (3–30 mg·kg⁻¹, i.p. and 10–56 mg·kg⁻¹, po) in the mouse four-plate model and decreased adjunctive drinking behaviour (56 mg·kg⁻¹, i.p.) in the rat scheduled-induced polydipsia model.

Conclusions and implications

These findings suggest that WAY-211612 may represent a novel antidepressant.     

A rare association of systemic lupus erythematosus, morbid obesity and Takotsubo Syndrome

The authors report a 68-year-old white female with long-term systemic lupus erythematosus as well as morbid obesity, characterized by very elevated body mass index, who presented a classical picture of acute coronary failure ascribed to Takotsubo syndrome.     

膳食补充剂黑醋与红曲单用及联用对中老年人血脂的影响

目的:观察并比较黑醋与红曲单用及联用对中国中老年人降低血脂的效能。方法:试验于2003-10/11在大连医科大学中日合作医药科学研究所进行。选择30名年龄为45￣65岁的中老年人。30名受试者被随机分为3组,每组10人,分别服用黑醋丸6粒,红曲丸6粒或黑醋 红曲丸6粒,1次/d,连续4周;3种膳食补充剂黑醋丸、红曲丸及黑醋 红曲丸由含有不同剂量的黑醋固形物、红曲粉末K-F、柠檬酸钠、大豆油、蜂蜡、脂肪酸甘油脂组成(由日本国第一药品工业株式会社提供)。各组服用剂量分别为每天黑醋固形物360mg,红曲粉72mg及黑醋固形物360mg 红曲粉72mg。服用2和(或)4周后,检测血脂和血糖,同时用血清样品检测肝功与肾功能。实验结果用SPSS统计软件进行方差分析。结果:30名中老年人全部进入结果分析。①受试者连续3d每日营养素摄入量:3组之间热量、胆固醇、膳食纤维、宏量或微量营养素摄入量差距均无显著性。②血清脂质浓度和计算的脂质比:2,4周血清三酰甘油含量,黑醋 红曲组低于基础值(分别下降(24.6±4.9)%,(27.6±3.4)%,P<0.05);红曲组与基础值比较,差异无显著性[分别下降(6.5±3.4)%,(11.6±4.5)%];黑醋组与基础值比较,差异无显著性[分别下降(10.2±7.3)%,(12.4±8.2)%]。2,4周血清胆固醇、低密度脂蛋白胆固醇含量,黑醋 红曲丸、红曲丸和黑醋丸组均比基础值降低,差异无统计学意义。3种膳食补充剂对血清高密度脂蛋白胆固醇及计算的脂质比值无明显改变。血糖含量无变化。未发现对受试对象产生明显不良反应。结论:服用黑醋 红曲丸能明显降低受试者血清三酰甘油含量。黑醋与红曲合用对高脂血症者可能有健康效益。     

子宫内膜异位症患者血清细胞因子水平的测定

子宫内膜异位症患者血清细胞因子水平的测定程凯灵令狐华子宫内膜异位症（ＥＭＳ）是妇科常见病。国内外学者的研究表明，免疫学的变化在该病的病理生理学中有重要意义。本研究通过酶联免疫吸附试验（ＥＬＩＳＡ），对ＥＭＳ患者血清细胞因子［白细胞介素（ＩＬ）－６、Ｉ...