Effects of large brain stem lesions on the cholinergic system in the rat cochlear nucleus

Large lesions were made medial to one cochlear nucleus in rats, in order to cut virtually all centrifugal pathways to it. To estimate the contribution of these centrifugal pathways to cholinergic synapses in the cochlear nucleus, choline acetyltransferase and acetylcholinesterase activities were mapped, by quantitative histochemical procedures, in lesion and control side cochlear nuclei. Choline acetyltransferase activities were reduced by 85–90% in most regions of the lesion side cochlear nucleus and by 65–75% in granular regions. Acetylcholinesterase activities were reduced by 50% or less in the same regions. The choline acetyltransferase results are consistent with a conclusion that by far most cholinergic synapses in the rat cochlear nucleus derive from centrifugal pathways. Additionally, the effects of the lesions on enzyme activities in the lateral superior olivary nucleus and ventral nucleus of the trapezoid body, and in the facial, motor trigeminal, and spinal trigeminal nuclei were examined. In the lesion side facial nucleus, 60% and 40% decreases in choline acetyltransferase and acetylcholinesterase activities, respectively, were apparently consequences of facial root transection. Lesion-control enzyme activity differences in the other nuclei were much smaller.     

Heterogeneity of motor units activating single Golgi tendon organs in cat leg muscles

Summary The aim of this study was to investigate whether an individual Golgi tendon organ can signal the contraction of motor units with different physiological properties. The axonal conduction velocity and tetanic tension of motor units were examined in four muscles of the cat leg (peroneus brevis, peroneus longus, tibialis anterior and soleus). The motor units which were found to activate a given tendon organ had contractile properties dispersed over the same range as those of the whole muscle population. The proportion of tendon organ-activating motor units found in the studied samples suggests that altogether, the Golgi tendon organs of a muscle monitor the contraction of every motor unit in this muscle.     

Behavioral naltrexone therapy: an integrated treatment for opiate dependence

Treatment of opiate dependence with naltrexone has been limited by poor compliance. Behavioral Naltrexone Therapy (BNT) was developed to promote adherence to naltrexone and lifestyle changes supportive of abstinence, by incorporating components from empirically validated treatments, including Network Therapy with a significant other to monitor medication compliance, the Community Reinforcement Approach, and voucher incentives. An overview is presented of the BNT treatment manual. In an uncontrolled Stage I trial (N = 47), 19% completed the 6-month course of treatment. Retention was especially poor in the subsample of patients who were using methadone at baseline (N = 18; 39% completed 1 month, none completed 6 months), and more encouraging among heroin-dependent patients (N = 29; 65% completed 1 month, 31% completed 6 months). Thus, attrition continues to be a serious problem for naltrexone maintenance, although further efforts to develop interventions such as BNT are warranted.     

Proinsulin 2 knockout NOD mice: a model for genetic variation of insulin gene expression in type 1 diabetes

Insulin is a major disease determinant in type 1 diabetes, type 2 diabetes, and related disorders. The role of variations in the expression of the insulin gene has been proposed in genetic susceptibility to the three pathological conditions in humans. In contrast to humans, rodents express two proinsulin isoforms. One isoform, proinsulin 1, is expressed exclusively in islets. The second, proinsulin 2, is expressed in islets and in other tissues, especially the thymus. We took advantage of the expression of these two isoforms to introduce a null proinsulin 2 allele in NOD mice and to evaluate the consequence of a variation of proinsulin 2 gene expression on the development of type 1 diabetes on the NOD genetic background. Heterozygote NOD mutant mice carrying a null proinsulin 2 mutation showed an increased incidence of type 1 diabetes at successive backcross generations. Plasma glucose and insulin levels were identical in prediabetic mutant and in wild-type mice at 4 weeks of age. Variation in insulin gene expression is hypothesized to interfere with diabetes development at both the islet and the thymus level.     

Histochemical localization of endometrial insulin-like growth factor binding protein-1 and -3 during the luteal phase in controlled ovarian hyperstimulation cycles: a controlled study

OBJECTIVE: To determine if controlled ovarian hyperstimulation (COH) affects the endometrial expression of IGFBP-1 and IGFBP-3. DESIGN: Prospective, controlled study. SETTING: Tertiary infertility clinic. PATIENT(S): Eighteen oocyte donors undergoing COH cycles and 17 natural cycle controls. INTERVENTION(S): Controlled ovarian hyperstimulation, endometrial biopsies. MAIN OUTCOME MEASURE(S): Immunohistochemical scoring of endometrial IGFBP-1 and -3 expression, morphological endometrial dating, and serum estradiol (E(2)), LH, and progesterone (P(4)) concentrations. RESULT(S): No statistically significant difference was observed between natural and stimulated cycles in change in IGFBP-1 or -3 over standardized cycle days throughout the window of embryo implantation (days 17-24). The IGFBP-1 and -3 expression was zero or near zero for both the natural and COH cycles until day 12-13. Both IGFBPs showed increased production throughout the secretory phase. Advanced endometrial histology (>/=1 day) in glands and stroma was noted in COH cycles. Significant positive correlations of E(2) and P(4) were noted with IGFBP-1 and -3 but not with advanced endometrial morphology in the COH cycles. CONCLUSION(S): The COH cycles have no significantly increased endometrial IGFBP-1 or -3 expression throughout the implantation phase of the luteal cycle compared with normal menstrual cycles. Both IGFBPs were absent in the proliferative phase and increased throughout the secretory portion of the embryo implantation window.     

Retraction Note: Patients with ankylosing spondylitis treatment by golimumab: a systematic review and meta‑analysis

European Spine Journal -     

Genetic engineering of hematopoietic stem cells to generate invariant natural killer T cells

Invariant natural killer T (iNKT) cells comprise a small population of αβ T lymphocytes. They bridge the innate and adaptive immune systems and mediate strong and rapid responses to many diseases, including cancer, infections, allergies, and autoimmunity. However, the study of iNKT cell biology and the therapeutic applications of these cells are greatly limited by their small numbers in vivo (∼0.01–1% in mouse and human blood). Here, we report a new method to generate large numbers of iNKT cells in mice through T-cell receptor (TCR) gene engineering of hematopoietic stem cells (HSCs). We showed that iNKT TCR-engineered HSCs could generate a clonal population of iNKT cells. These HSC-engineered iNKT cells displayed the typical iNKT cell phenotype and functionality. They followed a two-stage developmental path, first in thymus and then in the periphery, resembling that of endogenous iNKT cells. When tested in a mouse melanoma lung metastasis model, the HSC-engineered iNKT cells effectively protected mice from tumor metastasis. This method provides a powerful and high-throughput tool to investigate the in vivo development and functionality of clonal iNKT cells in mice. More importantly, this method takes advantage of the self-renewal and longevity of HSCs to generate a long-term supply of engineered iNKT cells, thus opening up a new avenue for iNKT cell-based immunotherapy.Invariant natural killer T (iNKT) cells are a small population of αβ T lymphocytes highly conserved from mice to humans. Like conventional αβ T cells, iNKT cells are derived from hematopoietic stem cells (HSCs) and develop in the thymus. However, they differ from conventional T cells in several important aspects, including their display of NK cell markers, their recognition of glycolipid antigens presented by the nonclassical monomorphic major histocompatibility complex (MHC) molecule CD1d, and their expression of semi-invariant T-cell receptors (identical α chains paired with a limited selection of β chains) (1, 2). Despite their small numbers in vivo (∼0.1–1% in mouse blood and ∼0.01–1% in human blood), iNKT cells have been suggested to play important roles in regulating many diseases, including cancer, infections, allergies, and autoimmunity (3). When stimulated, iNKT cells rapidly release a large amount of effector cytokines like IFN-γ and IL-4, both as a cell population and at the single-cell level. These cytokines then activate various immune effector cells, such as natural killer (NK) cells and dendritic cells (DCs) of the innate immune system, as well as CD4 helper and CD8 cytotoxic conventional αβ T cells of the adaptive immune system via activated DCs (3, 4). Because of their unique activation mechanism, iNKT cells can attack multiple diseases independent of antigen and MHC restrictions, making them attractive universal therapeutic agents (3, 4). Notably, because of the capacity of effector NK cells and conventional αβ T cells to specifically recognize diseased tissue cells, iNKT cell-induced immune reactions result in limited off-target side effects (3, 4).Restricted by their extremely low numbers, both the study of iNKT cells and their clinical applications have been challenging. iNKT T-cell receptor (TCR) transgenic mice (5, 6) and iNKT induced pluripotent stem (iPS) cell-derived transgenic mice (7) provide valuable tools to study iNKT cell biology in mice, but these methods are both costly and time-consuming. In addition, approaches using transgenic mice have no direct clinical application. As an alternative, a TCR-engineered HSC adoptive transfer strategy could overcome these limitations and become clinically applicable. Since its demonstration in mice in the early 2000s, this HSC-engineered T-cell strategy has been widely used to successfully generate both mouse and human antigen-specific conventional αβ T cells in multiple mouse and humanized mouse models (8–13). Human clinical trials testing this strategy for treating melanoma are also ongoing (14). Based on these previous works and the scientific principle that iNKT cells follow a “TCR instruction” development path similar to that of conventional αβ T cells (15), we hypothesized that HSCs could be engineered to express iNKT TCR genes and be programmed to develop into clonal iNKT cells. In the present report, we demonstrated the feasibility of this new HSC-engineered iNKT cell approach in mice and provided evidence to support its therapeutic potential in a mouse melanoma lung metastasis model.     

Effectiveness of a Mass Media Campaign in Promoting HIV Testing Information Seeking Among African American Women

“Take Charge. Take the Test.” (TCTT), a media campaign promoting HIV testing among African American women, was piloted in Cleveland and Philadelphia from October 2006 to October 2007. This study assesses TCTT's effectiveness in promoting HIV testing information seeking among target audiences in each pilot city. The authors analyzed data on telephone hotlines promoted by the campaign and the www.hivtest.org Web site to examine trends in hotline calls and testing location searches before, during, and after the campaign. Cleveland hotline data were available from October 1, 2005, through February 28, 2008, for a total of 29 months (N = 126 weeks). Philadelphia hotline data were available from May 1, 2006, through February 28, 2008, for a total of 22 months (N = 96 weeks). The authors assessed the relation between market-level measures of the campaign's advertising activities and trends in hotline call volume and testing location searches. They found a significant relation between measures of TCTT advertising and hotline calls. Specifically, they found that increases in advertising gross ratings points were associated with increases in call volume, controlling for caller demographics and geographic location. The campaign had similar effects on HIV testing location searches. Overall, it appears the campaign generated significant increases in HIV information seeking. Results are consistent with other studies that have evaluated the effects of media campaigns on similar forms of information seeking. This study illustrates useful methods for evaluating campaign effects on information seeking with data on media implementation, hotline calls, and zip code–based searches for testing locations.