IP-10 contributes to the inhibition of mycobacterial growth in an ex vivo whole blood assay

Interferon-γ inducible protein 10 (IP-10), is a potent chemoattractant that promotes migration of monocytes and activated T-cells to inflammation foci. IP-10 is elevated in serum of patients with chronic hepatitis C virus (HCV) and tuberculosis (TB) infections, although it remains to be determined the contribution of IP-10 in restricting Mycobacterium tuberculosis (Mtb) replication. Here, we investigated the impact of IP-10 on mycobacteria replication using the ex vivo model of human whole-blood (WB) assay. In particular, we compared the levels of IP-10 upon infection with different Mtb clinical strains and species of non-tuberculous mycobacteria (NTM) and evaluated how IP-10 may contain bacterial replication. Interestingly, we observed that the inhibition of the host enzyme dipeptidyl peptidase IV (DPP-IV), which inactivates IP-10 through cleavage of two amino acids at the chemokine N-terminus, restricted mycobacterial persistence in WB, supporting the critical role of full length IP-10 in mediating an anti-Mtb response. Addition of recombinant IP-10 expressed in eukaryotic cells enhanced the anti-mycobacterial activity in WB, although no differences were observed when IP-10 containing different proportions of cleaved and non-cleaved forms of the chemokine were added. Moreover, recombinant IP-10 did not exert a direct anti-mycobacterial effect. Our results underscore the clinical relevance of IP-10 in mycobacteria pathogenesis and support the potential outcomes that may derive by targeting the IP-10/CXCR3 pathway as host directed therapies for the treatment of Mtb or NTM infections.     

Gemelli and obturator internus muscles: Different heads of one muscle?

Background: The superior gemellus, inferior gemellus, and obturator internus muscles were once regarded as a single muscle judging from their insertion and function. However, current textbooks of anatomy do not treat them as one muscle. In gross anatomy, the classification of muscles depends largely upon the nerve supply, so that the present author re-examined the nerve supply to the three muscles. Methods: Fourteen nerve-muscle specimens were taken from 12 cadavers (five males and seven females) and examined with the unaided eye and under a dissecting microscope. Results: (1) The modes of nerve supply to the superior gemellus, inferior gemellus, and obturator internus muscles differed; however, the nerves to the muscles shared the same spinal nerve components. (2) The gemelli formed a muscular pocket (“gemellus pocket”) through which obturator internus muscle passed. Conclusions: In light of this knowledge on nerve muscle relationships, the difference in the pattern of nerve supply to the superior gemellus, inferior gemellus, and obturator internus muscles cannot be the basis for stating that the muscle are independent. Rather, their fusion to form the gemellus pocket and their common insertion suggest that they are different heads of one muscle. © 1995 Wiley-Liss, Inc.     

The effect of ranitidine, alone and in combination with clemastine, on allergen-induced cutaneous wheal-and-flare reactions in human skin

The effect of intradermal ranitidine (administered alone and in combination with clemastine) on allergen-mediated wheal-and-flare reactions has been evaluated in a double-blind study on 10 healthy atopic volunteers. Ranitidine alone, administered in doses over a 10(4)-fold concentration range, had no effect on the size either of allergen-induced wheal or flare reactions. Clemastine alone evoked a dose-related inhibition of both wheal and flare. Compared to the inhibition achieved by clemastine alone, the combination of ranitidine with clemastine produced a small but significant increase in inhibition of allergen-induced flare at ranitidine concentrations of 10(-5) mol/L (p less than 0.001) and 10(-6) mol/L (p less than 0.01), and of allergen-induced wheal at ranitidine concentration 10(-5) mol/L (p less than 0.01). Our results provide further evidence for the presence of cutaneous histamine H2 receptors and their participation in the formation of allergen-mediated skin reactions but indicate that the contribution of cutaneous histamine H2-receptor stimulation to the production of immediate wheal-and-flare reactions evoked by allergen is only modest.     

Induction of suppressor cells, interleukin-2 production and mitogenesis with monomeric concanavalin a: Different actions of tetrameric and monomeric concanavalin A

The ability of a photoalkylated monomeric concanavalin A (Con A) derivative to induce mitogenesis, interleukin-2 (IL-2) production and suppressor cells in murine spleen cell cultures has been compared with the activity of native, tetrameric Con A. The monomeric derivative was prepared by photochemically induced alkylation of tryptophan residues of tetravalent Con A in the presence of chloroacetamide followed by sizing chromatography [Tanaka et al. (1981) J. Biochem. 89, 1643–1646]. The monomeric derivative appeared to display less mitogenic activity than the tetramer and was also less effective in inducing IL-2 production. No difference was detected between the monomeric and tetrameric forms of Con A in inducing suppressor cells. The data suggest that cross-linking and bridging via sugar-binding sites, while potentiating mitogenesis and IL-2 production, had little effect on suppressor cell induction.     

Attrition in maintenance therapy for recurrent depression. A preliminary report

All maintenance treatment programs are complicated by the issue of patient noncompliance. This report investigates factors contributing to noncompliance during a 2-year study designed to evaluate the efficacy of long-term antidepressant medication in patients with recurrent unipolar depression. Only 21 of 51 patients (49%) who entered maintenance treatment successfully completed this phase of the study. Fifteen patients (8 completers and 7 dropouts) were randomly selected for an interview which focused on their previous psychiatric treatment history and attitudes towards the maintenance treatment program. In addition, these patients also completed a comprehensive personality battery. Results indicate that, while both groups had similar attitudes about the treatment program, they differed significantly along personality and psychiatric treatment history variables. Dropouts scored higher than completers on a measure of hysterical personality style. They were also more likely to have received psychotherapy in previous treatment experiences and to rate it as beneficial, while completers consistently rated prior treatment, which did not include antidepressant medication, as being of no benefit whatsoever. In order to enhance patient compliance, it is important to obtain information early in treatment about patients' treatment histories and their expectations about effective treatment for depression.     

Fragile sites and cancer breakpoints

To determine whether there might be a statistically significant association between fragile sites and cancer breakpoints, we examined the locations of the 21 fragile sites and the 50 cancer breakpoints recently accepted by the Seventh Human Gene Mapping Workshop. Nine of the 21 fragile sites appeared to be located at or near a cancer breakpoint. The chi-square test for association gives a value of 15.8 (p < 0.001) indicating that there is a very highly significant statistical association between human fragile sites and cancer breakpoints. This association is not narrowly limited to one class of fragile site, such as those sensitive to folate or to one type of cancer, but appears to extend to leukemia, lymphoma, and solid cancer. To more fully understand the meaning of this intriguing association between fragile sites and cancer breakpoints, future research will need to locate additional fragile sites and cancer breakpoints with precision, record their concurrence in individuals and families, determine if fragile site families are predisposed to cancer, and prove that a fragile site and a cancer breakpoint that appear to be coincident are at the same point on the DNA level.     

Is transfusion-transmitted dengue fever a potential public health threat?

Dengue is an arboviruses due to single-stranded enveloped ribonucleic acid viruses, named dengue viruses (DENV), that include four serotypes and are mainly transmitted via the bite of mosquitoes of the genus Aedes (A. aegypti and A. albopictus). The distribution of the disease was historically limited to intertropical areas; however, during the last thirty years, the perimeter of the disease extended considerably and temperate areas are now at risk of outbreaks. The present global burden of dengue is considerable: 2.5 billion people over more than 100 countries are concerned; 50 to 100 million infections occur every year, with a number of fatal cases of approximately 20000. Although frequently asymptomatic or limited to a mild fever, dengue is responsible for severe cases mainly consecutive to the occurrence of hemorrhagic complications that can lead to shock and death, notably in children from poor-resource settings. The place of DENV as a transfusion-transmitted pathogen has been recognized only in 2008. At the present time, only five cases of transfusion-transmitted dengue, including one case of dengue hemorrhagic fever, have been formerly documented. This review provides a general overview of dengue, its viruses and their vectors. It replaces the disease in the context of other viral diseases transmitted by arthropods. It discusses the threat of dengue on the supply of blood products in endemic and non endemic areas. Finally, it describes the specific and non specific measures available for improving the security of blood products with regards to this emerging risk. Interestingly, in 2009, the American Association of Blood Banks placed DENV in the highest category of emerging infectious agents for their potential impact on transfusion recipient safety for the next years in North America.     

Early events and implication of F-actin and annexin I associated structures in the phagocytic uptake of Brucella suis by the J-774A.1 murine cell line and human monocytes

Brucella spp. are facultative, intracellular pathogenic bacteria that cause brucellosis, a zoonosis affecting mammalian species. Brucella entry into myelomonocytic cell lines is highly enhanced by opsonization. Few studies have been undertaken to unravel the first interactions between these bacteria and their host cells. This paper deals with early events following contact of Brucella suis with the J-774A.1 phagocytic cell line and differentiated monocytes. Phagocytic uptake of bacteria was documented under a fluorescence microscope using GFP-expressing B. suis. Unlike entry in the J-774A.1 cell line, non-opsonized Brucella entered differentiated human monocytes as efficiently as opsonized bacteria. However, following 1 h infections, a mean of only three bacteria were phagocytized and the whole monocyte population was only infected after a 4 h infection. Contact of non-opsonized Brucella with phagocytes did not induce marked structural changes at the cell surface, as revealed by scanning electron microscopy. Contact of Brucella (opsonized or not) elicited transient local recruitment of F-actin, revealed by phalloidin labelling, and of annexin I-associated structures, revealed by immunofluorescence staining. Finally, bacteria appeared to be rapidly internalized in monocytes once they had adhered to the cell surface. A low percentage of infected cells and few adhered and/or internalized bacteria following short-term infections could have resulted either from the fact that there were few sites of entry or the weak bacterial initial interactions with the host-cell membrane or the bacterial receptor.     

Comparison of four weeks'' treatment with fenoterol and terbutaline aerosols in adult asthmatics : A double-blind crossover study

Fenoterol and terbutaline, two long-acting beta 2-adrenoceptor agonists in aerosol form, were compared in an 8-wk randomized double-blind crossover study in 22 mild to moderately severe asthmatics. Patients completed diary cards, recorded peak expiratory flow rate (PEFR) twice daily, and attended a clinic for measurement of PEFR, 1-sec forced expiratory volume (FEV1), and forced vital capacity (FVC) twice during each treatment period. Fifteen patients completed the study; 5 dropped out while using fenoterol, and 2 while using terbutaline. At clinic attendances, the patients had a significantly higher mean PEFR after 4 wk on terbutaline (385 L/min) than after fenoterol (316 L/min) (p less than 0.001). Similar results were found on analysis of the morning and evening PEFR recordings. On comparing each individual's PEFR recordings during the 2 treatments, it was found that there was no significant difference among the treatments in 3 patients, while 9 patients had a better response to terbutaline, and 3 patients had a better response to fenoterol. While similar number expressed a subjective preference for each treatment, the lung function data suggested that the effectiveness of fenoterol appeared to decline during the trial period.     

Novel mutations in the ciliopathy-associated gene CPLANE1 (C5orf42) cause OFD syndrome type VI rather than Joubert syndrome

Mutations in CPLANE1 (previously known as C5orf42) cause Oral-Facial-Digital Syndrome type VI (OFD6) as well as milder Joubert syndrome (JS) phenotypes. Seven new cases from five unrelated families diagnosed with pure OFD6 were systematically examined. Based on the clinical manifestations of these patients and those described in the literature, we revised the diagnostic features of OFD6 and include the seven most common characteristics: 1) molar tooth sign, 2) tongue hamartoma and/or lobulated tongue, 3) additional frenula, 4) mesoaxial polydactyly of hands, 5) preaxial polydactyly of feet, 6) syndactyly and/or bifid toe, and 7) hypothalamic hamartoma. By whole or targeted exome sequencing, we identified seven novel germline recessive mutations in CPLANE1, including missense, nonsense, frameshift and canonical splice site variants, all causing OFD6 in these patients. Since CPLANE1 is also mutated in JS patients, we examined whether a genotype-phenotype correlation could be established. We gathered and compared 46 biallelic CPLANE1 mutations reported in 32 JS and 26 OFD6 patients. Since no clear correlation between paired genotypes and clinical outcomes could be determined, we concluded that patient's genetic background and gene modifiers may modify the penetrance and expressivity of CPLANE1 causal alleles. To conclude, our study provides a comprehensive view of the phenotypic range, the genetic basis and genotype-phenotype association in OFD6 and JS. The updated phenotype scoring system together with the identification of new CPLANE1 mutations will help clinicians and geneticists reach a more accurate diagnosis for JS-related disorders.     

Interdependence of measures in pavlovian conditioned freezing

Pavlovian conditioned freezing is an intensively utilized paradigm that has become a standard model of memory and cognition. Despite its widespread use, the interdependence among each measure commonly reported in fear conditioning studies has not been described. Using mice, we examine the relationship of each common freezing measure (Training Baseline, Post-Shock freezing, Contextual Fear, Tone Baseline, and Tone Fear), as well as baseline locomotor activity measures, to better understand the significance of each. Of particular interest, Post-Shock freezing appears to be a good measure of immediate contextual memory. In contrast, Tone Baseline freezing, as typically measured in a novel context, appears to be contaminated with multiple sources of fear. Finally, Contextual and Tone Fear show a weak interdependence.     

Validation of a Hamilton subscale for endogenomorphic depression

The current study examined the validity of a subscale for endogenomorphic depression derived from the Hamilton Rating Scale for Depression. In a sample of 147 women outpatients with primary depression, subscale (HES) scores were bimodally distributed around the mean score of 7.38. High-HES patients had significantly elevated scores on measures of depressive symptomatology, generalized symptomatic distress, and social impairment relative to low-HES patients. Classifications based on HES scores significantly predicted RDC and DSM-III subtype diagnoses of endogenous and nonendogenous depression. Diagnostic predictions based on the subscale's items were superior to predictions made using the 'non-endogenomorphic' Hamilton items. Potential applications for research are discussed.     

A modified epon embedding and counterstaining procedure for light microscopic examination of Golgi impregnated neurons in vertebrates

Tissue blocks impregnated with the Golgi-Kopsch method were embedded in a modified Epon formulation (1.5 parts DDSA, 1 part Epon 812, 1.9% DMP-30). Sections 150 μm thick were cut with a steel knife on a sliding microtome. The surface of the sections were counterstained with basic dyes to localize Golgi-impregnated neurons within cyto-architectonically defined areas. Neither the embedding nor counterstaining procedure appeared to adversely affect the quality of the Golgi preparation.     

Detection of virus and cellular-determined antigens in situ using [125I]protein A and autoradiography.

The present paper describes the use of [125I]Protein A, isolated from Staphylococcus aureus, in detecting antigen-antibody complexes by autoradiography on single cells. the method is relatively quick, reproducible, potentially more sensitive than immunofluorescence, and should be useful in combination with conventional radioimmuno-assays. We have used it to detect the cellular expression of IgM, kappa, lambda, and beta 2-microglobulin, as well as the expression of Epstein-Barr Virus (EBV)-associated antigens expressed in human lymphoblastoid cell lines.     

Raddeanin A inhibited epithelial-mesenchymal transition (EMT) and angiogenesis in glioblastoma by downregulating β-catenin expression

Raddeanin A (RA), an oleanane-type triterpenoid saponin derived from Anemone raddeana Regel, has been found to suppress the viability and metastasis of several cancers, including GBM, through various signaling pathways. However, the mechanisms underlying the anti-GBM properties of RA have not been fully elucidated. Epithelial to mesenchymal transition (EMT) and angiogenesis are important for the genesis and progression of GBM. These two crucial processes can be regulated by multiple molecular, including β-catenin, which has been demonstrated to act as a pro-tumorigenic molecular. In this study, we aimed to determine whether RA could suppress EMT and angiogenesis by inhibiting the action of β-catenin in GBM. We found that RA inhibited the proliferation, invasion and migratory properties of GBM cells. RA was also found to have downregulated the expressions of β-catenin and EMT-related biomarkers (N-cadherin, vimentin, and snail). In addition, the overexpression of β-catenin reversed the therapeutic effects of RA exerted on the EMT of GBM cells. RA restricted angiogenesis, as shown by the tube formation assay and CAM assay, while it downregulated VEGF levels in HUVECs. Moreover, massive β-catenin could reverse the suppression of angiogenesis induced by RA. Finally, we demonstrated that RA inhibited tumor growth and prolonged survival time in an intracranial U87 xenograft mouse model. Similar to the results in vitro, RA downregulated the expression of β-catenin, EMT makers and VEGF, and decreased vessel density in vivo. In summary, our results demonstrated that RA repressed GBM via downregulating β-catenin-mediated EMT and angiogenesis both in vitro and in vivo.     

Noninvolvement of chromosome 16 in karyotype evolution of acute myeloid leukemia in a patient with a heritable fragile site on 16q22

A fragile site on the long arm of chromosome #16 (q22) was detected in a 24-year-old man with pancytopenia. During the course of the disease he developed an inverted duplication of region q11-12 of chromosome #1 and a translocation between chromosomes #9 and #13: t(9;13)(p22;q32). These abnormalities, as well as an additional iso-like marker chromosome that consisted of one normal 9p and the abnormal 9p arm, were detected in Epstein-Barr nuclear antigen-positive B-cell cultures. Two years later, evolution of the abnormal clone with loss of chromosome #7 and, subsequently, chromosome #22 occurred in connection with development of acute myeloid leukemia. Although the heritable fragile site on chromosome #16 was present in all cell populations investigated, it was not involved in the evolution of the abnormal karyotype. This fragile chromosome #16 also was found in 4 of 11 family members in whom chromosome analysis was performed, thus suggesting this aberration was inherited in a dominant autosomal pattern. The incidence of the heritable fragile site in normal and leukemic cells of the patient, as well as stimulated blood cultures of his relatives, are reported. In addition, the possible relationship between this constitutional chromosome breakage syndrome and the occurrence of leukemia is analyzed.