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Este trabalho propõe a implementação e o monitoramento do plano de gerenciamento de resíduos em uma instituição, através da aplicação de auditorias e propostas de ações corretivas e preventivas, a fim de obter uma melhoria contínua no gerenciamento adequado dos resíduos. A metodologia PGR - Plano de Gerenciamento de Resíduos que une o ciclo de melhoria continua (PDCA) baseado nas etapas da Lei 12305/10 - Política Nacional de Resíduos Sólidos (PNRS), auditorias, políticas e legislações correlatas e ainda, o conceito 4 R’s (Reduzir, Reutilizar, Reciclar e Repensar) com a aplicação e objetivos e metas (MORAES et al, 2019), foi aplicada neste trabalho. Os estudos permitiram concluir que as auditorias são uma ferramenta de suma importância para o monitoramento de um Plano de Gerenciamento de Resíduos e para a implementação da PNRS, as quais devem ser realizadas anualmente, evitando penalidades e auxiliando no gerenciamento adequado dos resíduos sólidos e como subsídio para a gestão ambiental em instituições.

The effects of hydrogen on the ductility loss and fracture behavior of a nitrogen-doped duplex stainless steel were investigated via tensile testing after electrochemical hydrogen pre-charging. Post-mortem microstructure analyses were performed to characterize the microstructural damage evolution and fracture surface. Hydrogen charging of the steel resulted in quasi-cleavage and intergranular fractures, which were associated with transgranular austenite cracking and ferrite/austenite interface cracking, respectively. The crucial factors that resulted in brittle-like cracking were deformation twinning in austenite and nitrogen-related solution hardening.

Mycobacterium tuberculosis (M. tuberculosis) is the causative agent of tuberculosis in human. One of the major M. tuberculosis virulence factors is early secretory antigenic target of 6-kDa (ESAT-6), and EccB5 protein encoded by eccB5 is one of its components. EccB5 protein is a transmembrane protein in ESX-5 system. The aim of this study is to explore the characteristics of wild-type EccB5 and its mutant form N426I. We expressed the EccB5 protein by cloning the mutant and wild-type eccB5 gene in Escherichia coli (E. coli). We compared the protein structure of wild type and mutant form of EccB5 and found changes in structure around Asn426 (loop structure) in wild type and around Ile426 (β-strand) in the mutant. The truncated recombinant protein of EccB5 was successfully cloned and expressed using plasmid pCold I in E. coli DH5α and E. coli strain Rosetta-gami B (DE3) and purified as a 38.6 kDa protein by using the affinity column. There was no detectable adenosine triphosphatase activity in truncated forms of EccB5 and its mutant. In conclusion, our study reveals successful cloning and protein expression of truncated form of eccB5 gene of M. tuberculosis. EccB5 protein in ESX-5 system may be an important membrane component involved in the transport machinery of type VII secretion system, which is essential for growth and virulence.

Background Individuals with relapses of leprosy should be monitored carefully, however, with respect to paucibacillary (PB) leprosy, it is sometimes difficult to make a definitive diagnosis of relapse, because the bacillary index is often negative. To evaluate the usefulness of cytokine profiling in a patient with relapsed PB leprosy who tested negative for anti-phenolic glycolipid-I antibodies, we analyzed the Mycobacterium leprae protein-induced cytokine expression in peripheral blood mononuclear cells of the patient. Case presentation An 89-year-old-male relapsed PB patient, first treated for leprosy over 50 years prior, was examined. In April 2012, he noticed three skin lesions consisting of annular erythema in the thighs. Slit skin smear tests were negative, and skin biopsies revealed a pathology of indeterminate-to-borderline tuberculoid leprosy. He received 600 mg of rifampicin once per month and 75 mg of dapsone daily for 12 months. The annular erythemas disappeared after starting treatment. Before treatment, and 6 and 12 months after starting treatment, the Th1/Th2 cytokine profiles in the supernatant of mononuclear cells from the patient before and after stimulation with Mycobacterium leprae soluble protein (MLS) were examined using a Cytometric Bead Array (CBA) Human Th1/Th2 Cytokine Kit II. The CBA Enhanced Sensitivity Flex Set system was applied to detect small amounts of cytokines in the serum just before treatment and one year before relapse. In the culture supernatant, just before treatment, increases in IFN-γ level and the IFN-γ/IL-10 ratio and a decreased IL-6 level were observed without stimulation. Upon stimulation with MLS, just before treatment, both the IFN-γ and TNF levels increased markedly, and twelve months after starting treatment, the IFN-γ and TNF levels decreased greatly. In the serum, just before treatment, increases in IFN-γ and TNF levels and the IFN-γ/IL-10 ratio were evident compared with those measured one year before relapse. Conclusions Cytokine profiling using culture supernatants and serum samples may be useful for the diagnosis of relapsed PB leprosy.

Carpal Tunnel Syndrome (CTS) is a median nerve entrapment neuropathy that alters primary somatosensory cortex (S1) organization. While electro-acupuncture (EA), a form of peripheral neuromodulation, has been shown to improve clinical and neurophysiological CTS outcomes, the role of EA-evoked brain response during therapy (within and beyond S1) for improved outcomes is unknown. We investigated S1-associated whole brain fMRI connectivity during both a resting and sustained EA stimulation state in age-matched healthy controls (N = 28) and CTS patients (N = 64), at baseline and after 8 weeks of acupuncture therapy (local, distal, or sham EA). Compared to healthy controls, CTS patients at baseline showed decreased resting state functional connectivity between S1 and thalamic pulvinar nucleus. Increases in S1/pulvinar connectivity strength following verum EA therapy (combined local and distal) were correlated with improvements in median nerve velocity (r = 0.38, p = 0.035). During sustained local EA, compared to healthy controls, CTS patients demonstrated increased functional connectivity between S1 and anterior hippocampus (aHipp). Following 8 weeks of local EA therapy, S1/aHipp connectivity significantly decreased and greater decrease was associated with improvement in patients' functional status (r = 0.64, p = 0.01) and increased median nerve velocity (r = −0.62, p = 0.013). Thus, connectivity between S1 and other brain areas is also disrupted in CTS patients and may be improved following EA therapy. Furthermore, stimulus-evoked fMRI connectivity adds therapy-specific, mechanistic insight to more common resting state connectivity approaches. Specifically, local EA modulates S1 connectivity to sensory and affective processing regions, linked to patient function and median nerve health.

The present study concerns the cryogenic processing of metals with simultaneous analysis of x-ray diffraction in a synchrotron ring. The mechanical properties improvement related to cryogenic processing of metals is attributed to the partial suppression of dynamic recovery. Thus, commercially pure metals with different stacking fault energies (silver, copper and aluminum) were deformed by uniaxial tensile tests and characterized by in situ x-ray diffraction, at room (293 K) and cryogenic (77 K) temperatures. The cryogenic processing allows a simultaneous improvement in ductility and strength for silver and copper and an improvement in strength for aluminum. This difference in mechanical properties was investigated by means of variations in crystallite size, microstrain and also the amount and size of dimples on the fracture surface. The microstructural refinement at cryogenic temperatures shows a tendency related to the stacking fault energies.

Mycobacterium leprae (M. leprae) is the etiological agent of leprosy, and the skin lesions of lepromatous leprosy are filled with numerous foamy or xanthomatous histiocytes that are parasitized by M. leprae. Lipids are an important nutrient for the intracellular survival of M. leprae. In this study, we attempted to determine the intracellular lipid composition and underlying mechanisms for changes in host cell lipid metabolism induced by M. leprae infection. Using high-performance thin-layer chromatography (HPTLC), we demonstrated specific induction of triacylglycerol (TAG) production in human macrophage THP-1 cells following M. leprae infection. We then used [14C] stearic acid tracing to show incorporation of this newly synthesized host cell TAG into M. leprae. In parallel with TAG accumulation, expression of host glycerol-3-phosphate acyltransferase 3 (GPAT3), a key enzyme in de novo TAG synthesis, was significantly increased in M. leprae-infected cells. CRISPR/Cas9 genome editing of GPAT3 in THP-1 cells (GPAT3 KO) dramatically reduced accumulation of TAG following M. leprae infection, intracellular mycobacterial load, and bacteria viability. These results together suggest that M. leprae induces host GPAT3 expression to facilitate TAG accumulation within macrophages to maintain a suitable environment that is crucial for intracellular survival of these bacilli.

The pre-deformation effects of hydrogen embrittlement on nitrogen-doped duplex stainless steel were investigated using tensile testing. Hydrogen pre-charging and pre-straining were adopted, which increased the yield strength, but decreased the ductility. Silver decoration revealed that diffusible hydrogen was preferentially located in the ferrite or grain boundaries of the undeformed steel, whereas the 22% pre-strained steel had diffusible hydrogen in austenite. When deformed, the preferential plastic deformation path of the steel was in the ferrite; hence, the apparent diffusivity in ferrite decreased, while the austenite had a relatively high apparent hydrogen flux. Pre-straining suppressed macroscopic hydrogen diffusion and local diffusion in ferrite and austenite by increasing the dislocation density. The degree of the suppression of the local diffusion was more significant in ferrite than austenite. The changes in dislocation and hydrogen behaviors associated with pre-straining altered the cracking sites in the austenite and ferrite, and decreased hydrogen embrittlement susceptibility.

Carpal Tunnel Syndrome (CTS) is an entrapment neuropathy of the median nerve that alters the organization of the brain's primary somatosensory cortex (S1), both structurally and functionally. Our prior research also demonstrated that electro-acupuncture (EA), both local and remote/distal to the wrist, but not sham acupuncture, improved S1 somatotopy and peripheral (median) nerve conduction. However, the mechanisms linking EA-evoked brain response during therapy with improved clinical outcomes are unknown. We investigated functional connectivity during both rest and sustained electro-acupuncture at baseline in healthy controls and CTS patients and after 8 weeks of acupuncture therapy (local, distal, or sham EA) in CTS patients. Six minutes of 3T BOLD fMRI data were collected at rest and during EA. Data were processed with FMRIPREP and FSL to generate maps of connectivity between the S1 hand region and the rest of the brain (z > 2.3, pFWE

Mycobacterium bovis BCG has been the only practical vaccine for tuberculosis. However, BCG cannot fully prevent adult pulmonary tuberculosis. Therefore, the improvement of BCG vaccine is necessary. We previously produced recombinant (r) BCG (BCG-PEST) for the better control of tuberculosis. BCG-PEST was developed by introducing PEST-Heat Shock Protein (HSP)70-Major Membrane Protein (MMP)-II-PEST fusion gene into urease-deficient rBCG using antibiotic-resistant gene for the selection of rBCG. HSP70-MMPII fusion protein is highly immunogenic and PEST sequence was added to enhance processing of the fusion protein. Although BCG-PEST effectively inhibited intrapulmonary growth of Mycobacterium tuberculosis (MTB), BCG with antibiotic-resistant gene is not appropriate for human use. Therefore, we produced antibiotic-resistant gene-free rBCG. We generated leucine-biosynthetic gene (leuD)-deficient BCG and introduced the fusion gene with leuD as the selection marker and named this rBCG as BCG-LeuPH. BCG-LeuPH activated human naïve T cells of both CD4 and CD8 subsets and efficiently inhibited aerosol-challenged MTB in mice. These results indicate that leuD can replace antibiotic-resistant gene for the selection of vaccine candidates of rBCG for human use.

Enhancement of the T cell-stimulating ability of Mycobacterium bovis BCG (BCG) is necessary to develop an effective tuberculosis vaccine. For this purpose, we introduced the PEST-HSP70-major membrane protein-II (MMPII)-PEST fusion gene into ureC-gene depleted recombinant (r) BCG to produce BCG-PEST. The PEST sequence is involved in the proteasomal processing of antigens. BCG-PEST secreted the PEST-HSP70-MMPII-PEST fusion protein and more efficiently activated human monocyte-derived dendritic cells (DCs) in terms of phenotypic changes and cytokine productions than an empty-vector-introduced BCG or HSP70-MMPII gene-introduced ureC gene-depleted BCG (BCG-DHTM). Autologous human naive CD8+ T cells and naive CD4+ T cells were effectively activated by BCG-PEST and produced IFN-γ in an antigen-specific manner through DCs. These T cell activations were closely associated with phagosomal maturation and intraproteasomal protein degradation in antigen-presenting cells. Furthermore, BCG-PEST produced long-lasting memory-type T cells in C57BL/6 mice more efficiently than control rBCGs. Moreover, a single subcutaneous injection of BCG-PEST more effectively reduced the multiplication of subsequent aerosol-challenged Mycobacterium tuberculosis of the standard H37Rv strain and clinically isolated Beijing strain in the lungs than control rBCGs. The vaccination effect of BCG-PEST lasted for at least 6 months. These results indicate that BCG-PEST may be able to efficiently control the spread of tuberculosis in human.

Objective The aim of this study was to establish reference values for the width of the interval between the anterior and middle scalene muscles using ultrasonography during varying degrees of glenohumeral joint (GH) abduction. Reliability and body mass index (BMI) data were also assessed. Methods Interscalene triangles of asymptomatic participants were scanned bilaterally in the transverse plane. Images were obtained at 0°, 90°, and 150° of GH abduction with the participant seated. Width measurements were taken between the anterior and middle scalene muscle borders by bisecting the C6 nerve root as it passed superficial to the posterior tubercle of the C7 transverse process. Intra- and interexaminer reliability and BMI correlation were studied. Statistical significance was defined as P ≤ .05. Results Images of 42 scalene intervals were included from 21 participants (11 female). Mean participant age was 25.3 ± 3.9 years; mean BMI was 25.4 ± 2.7 kg/m 2 . Scalene interval measurements at 0°, 90°, and 150° of GH abduction were 4.5 ± 0.5 mm, 4.6 ± 0.5 mm, and 4.4 ± 0.7 mm, respectively, without a significant difference ( P = .07). Intraexaminer reliability was excellent (0°: intraclass correlation coefficient [ICC] = 0.82; 90°: ICC=0.89; 150°: ICC=0.90). Interexaminer reliability was good to excellent (0°: ICC=0.59; 90°: ICC=0.85; 150°: ICC=0.89). Body mass index was positively correlated only at 0° of GH abduction. Conclusions This study establishes previously unreported reference ultrasonography values for the width of the scalene interval. Intraexaminer reliability was excellent at all glenohumeral positions, and interexaminer reliability was determined to be good to excellent. Body mass index was positively correlated only at 0° of GH abduction.

Background and Aim CpG motifs in DNA sequences are recognized by TLR9 and activate immune cells. Bacterial genomic DNA (gDNA) has modified cytosine bases (5-methylcytosine (5mC)) and modified adenine bases (6-methyladenine (6mA)). It is well-known that 5mC inhibits immune activation by CpG DNA; however, it is still unclear whether 6mA inhibits immune activation by CpG DNA. In this study, we tried to examine whether the existence of 6mA affected the immunostimulatory activity of gDNA. Materials and Methods We focused on adenine methyltransferases (MTases) expressed by Mycobacterium bovis BCG (BCG). Each adenine MTases has specific target sites. Therefore, we used oligodeoxynucleotides (ODNs) with 6mA on the target sequences of MTases from BCG. We stimulated murine bone-marrow derived macrophages (BMDMs) with ODNs harboring 6mA or ODNs without 6mA. We then compared the immunostimulatory activity of ODNs with 6mA to ODNs without 6mA using. We also developed adenine MTase-deficient BCG. We analyzed whether 6mA on gDNA from BCG is involved in the immunostimulatory activity of BCG gDNA. Results Our results showed that 6mA located at the target sequence of mamA, an adenine MTase from BCG, enhanced IL-12p40 production from BMDMs stimulated with CpG DNA. We also observed that gDNA from mamA-deficient BCG induced less IL-12p40 from BMDMs than gDNA from mamA-intact BCG. Conclusion We concluded from these results that adenine methylation on ODNs may enhance immune activity induced by bacterial gDNA.

CpG motifs in DNA sequences are recognized by Toll-like receptor 9 and activate immune cells. Bacterial genomic DNA (gDNA) has modified cytosine bases (5-methylcytosine [5 mC]) and modified adenine bases (6-methyladenine [6 mA]). 5 mC inhibits immune activation by CpG DNA; however, it is unclear whether 6 mA inhibits immune activation by CpG DNA. Mycobacterium bovis BCG (BCG) has three adenine methyltransferases (MTases) that act on specific target sequences. In this study, we examined whether the 6 mA at the target sites of adenine MTases affected the immunostimulatory activity of CpG DNA. Our results showed that only 6 mA located at the target sequence of mamA, an adenine MTase from BCG, enhanced interleukin (IL)-12p40 production from murine bone marrow-derived macrophages (BMDMs) stimulated with CpG DNA. Enhancement of IL-12p40 production in BMDMs was also observed when BMDMs were stimulated with CpG DNA ligated to oligodeoxynucleotides (ODNs) harboring 6 mA. Accordingly, we then evaluated whether gDNA from adenine MTase-deficient BCG was less efficient with regard to stimulation of BMDMs. Indeed, gDNA from a mamA-deficient BCG had less ability to activate BMDMs than that from wild-type BCG. We concluded from these results that adenine methylation on ODNs and bacterial gDNA may enhance immune activity induced by CpG DNA.

Oral manifestation of diffuse large B-cell lymphoma (DLBCL) usually represents late involvement of disseminated and advanced disease. In this case, the oral lesion was the first sign of the disease. A 66-year-old male patient complained of difficulty adapting the upper total prosthesis for the last month. No underlying diseases have been reported except recent fatigue and weight loss complaint. Physical examination revealed a submucosal tumor in the maxillary region, which was 4 cm, exophytic, painless, and of fibrous consistency, and mucosa of recoating with areas of ulceration. Computed tomography showed an infiltrative, unilocular, isodense, expansive lesion with maxillary bone alteration as well as multiple bilateral supra and infradiaphragmatic lymphadenopathy. An incisional biopsy of the lesion was performed, and the morphologic profile associated with the immunohistochemical profile favored the diagnosis of DLBCL. The patient was submitted to the rituximab, cyclophosphamide, doxorubicin (hydroxydaunorubicin), vincristine (Oncovin), and prednisone regimen associated with intrathecal chemotherapy with complete remission of the oral lesion.

More than 100 counties, mainly in southwest China, report incidence rates of leprosy >1/100,000. The current study analysed the epidemiology of leprosy in southwest China to improve our understanding of the transmission pattern and improve control programs. 207 counties were selected in southwest China. Leprosy patients and their household contacts were recruited. The data from the medical interview and the serological antileprosy antibody of the leprosy patients were analysed. A total of 2,353 new cases of leprosy were interviewed. The distribution of leprosy patients was partly associated with local natural and economic conditions, especially several pocket areas. A total of 53 from 6643 household contacts developed leprosy, and the incidence rate of leprosy in the household contacts was 364/100,000 person-years. We found that NDO-BSA attained higher positive rates than MMP-II and LID-1 regardless of clinical types, disability and infection time in leprosy patients. By means of combination of antigens, 88.4% patients of multibacillary leprosy were detected, in contrast to 59.9% in paucibacillary leprosy. Household contacts should be given close attention for the early diagnosis, disruption of disease transmission and precise control. Applications of serology for multi-antigens were recommended for effective coverage and monitoring in leprosy control.

Sustained elimination of leprosy as a global health concern likely requires a vaccine. The current standard, BCG, confers only partial protection and precipitates paucibacillary (PB) disease in some instances. When injected into mice with the T helper 1 (Th1)-biasing adjuvant formulation Glucopyranosyl Lipid Adjuvant in stable emulsion (GLA-SE), a cocktail of three prioritized antigens (ML2055, ML2380 and ML2028) reduced M. leprae infection levels. Recognition and protective efficacy of a single chimeric fusion protein incorporating these antigens, LEP-F1, was confirmed in similar experiments. The impact of post-exposure immunization was then assessed in nine-banded armadillos that demonstrate a functional recapitulation of leprosy. Armadillos were infected with M. leprae 1 month before the initiation of post-exposure prophylaxis. While BCG precipitated motor nerve conduction abnormalities more rapidly and severely than observed for control infected armadillos, motor nerve injury in armadillos treated three times, at monthly intervals with LepVax was appreciably delayed. Biopsy of cutaneous nerves indicated that epidermal nerve fiber density was not significantly altered in M. leprae-infected animals although Remak Schwann cells of the cutaneous nerves in the distal leg were denser in the infected armadillos. Importantly, LepVax immunization did not exacerbate cutaneous nerve involvement due to M. leprae infection, indicating its safe use. There was no intraneural inflammation but a reduction of intra axonal edema suggested that LepVax treatment might restore some early sensory axonal function. These data indicate that post-exposure prophylaxis with LepVax not only appears safe but, unlike BCG, alleviates and delays the neurologic disruptions caused by M. leprae infection., Leprosy: Working towards a safe and effective vaccine A leprosy vaccine candidate has been developed that raises immune responses against targets gleaned from naturally resistant individuals. Researchers from the United States and Japan, led by Malcolm Duthie, of Seattle’s Infectious Disease Research Institute, tested a Mycobacterium leprae vaccine candidate that generated immune responses mimicking those found in partially-resistant patients, and immune co-inhabitants of the severely infected. The candidate, dubbed LepVax, inhibited infection in mice and, when administered post-infection, delayed and mitigated nerve damage in armadillos. This contrasts with the current vaccine, BCG, which can precipitate leprosy symptoms when given after infection. This study also revealed that M. leprae infection can induce ‘silent’ pre-clinical nerve aberations. High-risk populations may already be infected with M. leprae, making safe and effective post-exposure prophylaxis a landmark step in combating both the individual and global burden of leprosy.

Sustained elimination of leprosy as a global health concern likely requires a vaccine. The current standard, BCG, confers only partial protection and precipitates paucibacillary (PB) disease in some instances. When injected into mice with the T helper 1 (Th1)-biasing adjuvant formulation Glucopyranosyl Lipid Adjuvant in stable emulsion (GLA-SE), a cocktail of three prioritized antigens (ML2055, ML2380 and ML2028) reduced M. leprae infection levels. Recognition and protective efficacy of a single chimeric fusion protein incorporating these antigens, LEP-F1, was confirmed in similar experiments. The impact of post-exposure immunization was then assessed in nine-banded armadillos that demonstrate a functional recapitulation of leprosy. Armadillos were infected with M. leprae 1 month before the initiation of post-exposure prophylaxis. While BCG precipitated motor nerve conduction abnormalities more rapidly and severely than observed for control infected armadillos, motor nerve injury in armadillos treated three times, at monthly intervals with LepVax was appreciably delayed. Biopsy of cutaneous nerves indicated that epidermal nerve fiber density was not significantly altered in M. leprae-infected animals although Remak Schwann cells of the cutaneous nerves in the distal leg were denser in the infected armadillos. Importantly, LepVax immunization did not exacerbate cutaneous nerve involvement due to M. leprae infection, indicating its safe use. There was no intraneural inflammation but a reduction of intra axonal edema suggested that LepVax treatment might restore some early sensory axonal function. These data indicate that post-exposure prophylaxis with LepVax not only appears safe but, unlike BCG, alleviates and delays the neurologic disruptions caused by M. leprae infection.

Aluminum, copper and silver samples, all of them face-centered cubic (FCC) metals, were rolled at room and cryogenic temperatures until equivalent strains (ε) were between 3.23 and 4.13. The cryogenic temperature (CT) and room temperature (RT) rolled samples were evaluated by hardness tests and X-ray diffraction (XRD), which indicate influence of stacking fault energy (SFE) on process. Lower SFE metals tend to exhibit dislocation densities significantly increased and as consequence, hardness too. It was also noted that after sometime exposed to RT, the materials rolled at CT present hardness decrease.

Antibodies to phenolic glycolipid (PGL)-I and major membrane protein (MMP)-II were evaluated for serodiagnosis of leprosy in Southwest China, and the role in predicting the occurrence of the disease in household contacts (HHCs) of leprosy was examined. Using PGL-I (natural disaccharide-octyl-bovine serum albumin) antigen-based diagnosis (IgM antibodies), we could detect 94.9% of multibacillary (MB) leprosy and 38.9% paucibacillary (PB) leprosy patients, whereas using MMP-II (IgG antibody), 88.1% of MB and 61.1% of PB patients were positive. By combining the 2 tests and considering either test positive as positive, 100% of MB patients and 72.2% of PB patients were found to test positive. Of the HHCs of leprosy, 28.3% and 30% had positive levels of PGL-I and MMP-II Abs, respectively. Seven out of 21 HHCs, who had high Ab titer to either antigen, developed leprosy during the follow-up period of 3 years. These data suggest that the measurement of both anti-PGL-I as well as anti-MMP-II antibodies could facilitate early detection of leprosy.

In the present paper a preliminary study was performed on the influence of mechanical milling on the synthesis of niobium nitrade. The niobium metal powder sample, passing # 635 mesh sieve, was processed by mechanical milling in SPEX mill for 8 hours using a ball-to-powder ratio of 7:1 and a nitrogen atmosphere. The powder was annealed at different temperatures, 900 °C, 1000 °C, 1100 °C and 1200 °C for 1 hour in a hydrogen and argon atmosphere to study their crystallization, which then were formed into blanks for analysis of the compressibility curves. These samples were also subjected to X-ray diffraction and the data were compared between the annealing temperatures. The compressibility curves of niobium samples with and without grinding were also evaluated, showing high compacting capacity. These samples were subjected to X-ray diffraction and X-ray fluorescence. As the formation of nitrides (Nb2N) was observed in SPEX type mill, the interest in studying the synthesis of nitrides came up, using mechanical milling in Attritor type mill. Same previous results of Attritor processing indicate a Nb2N and NbN synthesis after annealing treatments.

We have previously shown that the serodiagnosis using major membrane protein-II (MMP-II) is quite efficient in diagnosing leprosy. However, the detection rate of pauci-bacillary (PB) leprosy patients is still low. In this study, we examined the usefulness of major membrane protein-I (MMP-I) from Mycobacterium leprae. The MMP-I-based serodiagnosis did not show significantly high detection rate. However, when the mixture of MMP-I and MMP-II antigens was used, we detected 94.4% of multi-bacillary leprosy and 39.7% of PB patients. There were little correlation between the titers of anti-MMP-I antibodies (Abs) and that of anti-MMP-II Abs in PB patients' sera. Ten out of 46 MMP-II-negative PB leprosy patients were MMP-I positive, so that the detection rate of PB leprosy patient increased from 39.7% to 53.8% by taking either test positive strategy. We concluded that MMP-I can complement the MMP-II-based serodiagnosis of leprosy.

The purpose of this study was to investigate the reliability of the Goutallier classification system (GCS) for grading muscle fatty degeneration in the lumbar multifidus (LM) using magnetic resonance imaging (MRI) examinations.Lumbar spine MRI scans were obtained retrospectively from the radiology department imaging system. Two examiners (a chiropractic diagnostic imaging resident and a board certified chiropractic radiologist with 30 years of experience) independently graded each LM at the L4/5 and L5/S1 intervertebral level. ImageJ pixel analysis software (version 1.47; National Institutes of Health, Bethesda, MD) was used independently by 2 observers to quantify the percent fat of the LM and allow correlation between LM percent fat and GCS grade. Twenty-five subject MRIs were randomly selected. Magnetic resonance imaging scans were included if they were obtained using a 1.5 T imaging system and were excluded if there was evidence of spinal infection, tumor, fracture, or postoperative changes. For all tests, P.05 was defined as significant.Intraobserver reliability grading LM fat ranged from a weighted κ (κw) of 0.71 to 0.93. Mean interobserver reliability grading LM fat was κ(w), 0.76 to κ(w), 0.85. There was a significant (P.001) correlation between LM percent fat and GCS grade. Furthermore, interobserver reliability in determining percent fat was between intraclass correlation coefficient, 0.73 to intraclass correlation coefficient, 0.90.In this study, the GCS was reliable in grading LM fatty degeneration and correlated positively with a quantified percent fat value. In addition, ImageJ software (National Institutes of Health) was reliable between raters when quantifying LM percent fat.

For the purpose of obtainingMycobacterium bovisbacillus Calmette-Guérin (BCG) capable of activating human naive T cells, urease-deficient BCG expressing a fusion protein composed ofMycobacterium tuberculosis-derived major membrane protein II (MMP-II) and heat shock protein 70 (HSP70) of BCG (BCG-DHTM) was produced. BCG-DHTM secreted the HSP70-MMP-II fusion protein and effectively activated human monocyte-derived dendritic cells (DCs) by inducing phenotypic changes and enhanced cytokine production. BCG-DHTM-infected DCs activated naive T cells of both CD4 and naive CD8 subsets, in an antigen (Ag)-dependent manner. The T cell activation induced by BCG-DHTM was inhibited by the pretreatment of DCs with chloroquine. The naive CD8+T cell activation was mediated by the transporter associated with antigen presentation (TAP) and the proteosome-dependent cytosolic cross-priming pathway. Memory CD8+T cells and perforin-producing effector CD8+T cells were efficiently produced from the naive T cell population by BCG-DHTM stimulation. Single primary infection with BCG-DHTM in C57BL/6 mice efficiently produced T cells responsive toin vitrosecondary stimulation with HSP70, MMP-II, andM. tuberculosis-derived cytosolic protein and inhibited the multiplication of subsequently aerosol-challengedM. tuberculosismore efficiently than did vector control BCG. These results indicate that the introduction of MMP-II and HSP70 into urease-deficient BCG may be useful for improving BCG for control of tuberculosis.

Background Migraine is a neurovascular disorder in which altered functional connectivity between pain-modulating circuits and the limbic system may play a role. Cortical spreading depression (CSD), which underlies migraine aura (MWA), induces C-fos expression in the amygdala. The role of CSD and amygdala connectivity in migraine without aura (MwoA) is less clear and may differentiate migraine from other chronic pain disorders. Methods Using resting-state functional MRI, we compared functional connectivity between the amygdala and the cortex in MWA and MWoA patients as well as in healthy subjects and in two other chronic pain conditions not associated with CSD: trigeminal neuralgia (TGN) and carpal tunnel syndrome (CTS). Results Amygdala connectivity in both MWA and MWoA was increased to the visceroceptive insula relative to all other groups examined. Conclusion The observed increased connectivity within the limbic/viscerosensory network, present only in migraineurs, adds to the evidence of a neurolimbic pain network dysfunction and may reflect repetitive episodes of CSD leading to the development of migraine pain.

Objective Carpal tunnel syndrome (CTS) is a common median nerve entrapment neuropathy characterized by pain, paresthesias, diminished peripheral nerve conduction velocity (NCV) and maladaptive functional brain neuroplasticity. We evaluated structural reorganization in brain gray matter (GM) and white matter (WM) and whether such plasticity is linked to altered median nerve function in CTS. Methods We performed NCV testing, T1-weighted structural MRI, and diffusion tensor imaging (DTI) in 28 CTS and 28 age-matched healthy controls (HC). Voxel-based morphometry (VBM) contrasted regional GM volume for CTS versus HC. Significant clusters were correlated with clinical metrics and served as seeds to define associated WM tracts using DTI data and probabilistic tractography. Within these WM tracts, fractional anisotropy (FA), axial (AD) and radial (RD) diffusivity were evaluated for group differences and correlations with clinical metrics. Results For CTS subjects, GM volume was significantly reduced in contralesional S1 (hand-area), pulvinar and frontal pole. GM volume in contralesional S1 correlated with median NCV. NCV was also correlated with RD and was negatively correlated with FA within U-fiber cortico-cortical association tracts identified from the contralesional S1 VBM seed. Conclusions Our study identified clear morphometric changes in the CTS brain. This central morphometric change is likely secondary to peripheral nerve pathology and altered somatosensory afference. Enhanced axonal coherence and myelination within cortico-cortical tracts connecting primary somatosensory and motor areas may accompany peripheral nerve deafferentation. As structural plasticity was correlated with NCV and not symptomatology, the former may be a better determinant of appropriate clinical intervention for CTS, including surgery., Highlights ► CTS demonstrated reduced GM volume in S1, pulvinar, and frontal pole. ► GM volume in S1 was significantly correlated with median nerve conduction velocity. ► Median nerve velocity was linked to WM microstructure in tracts connected to S1.

We describe a case of bloodstream infection (BSI) caused by Campylobacter lari in a 58-year-old man diagnosed with lumbar pyogenic spondylitis. Anaerobic blood cultures, taken on the day of admission and on hospital day 4, were positive after 30 h of incubation, although no bacteria were detected by Gram staining. After subculture on 5 % sheep blood agar for 2 days at 35 °C in a 5 % CO2 environment, capnophilic, curved, gram-negative bacteria were recovered. The bacteria were identified as C. lari using a combination of phenotypic identification methods and partial 16S rRNA gene sequencing. The BSI was eradicated following combination therapy with intravenous tazobactam/piperacillin, oral erythromycin, and sulfamethoxazole/trimethoprim. These results suggest that accurate identification, to the species level, is important to determine effective treatment of BSI caused by Campylobacter spp. and can help us to understand the epidemiology.

Mycobacterium leprae is the causative agent of leprosy and also known to possess unique features such as inability to proliferate in vitro. Among the cellular components of M. leprae, various glycolipids present on the cell envelope are well characterized and some of them are identified to be pathogenic factors responsible for intracellular survival in host cells, while other intracellular metabolites, assumed to be associated with basic physiological feature, remain largely unknown. In the present study, to elucidate the comprehensive profile of intracellular metabolites, we performed the capillary electrophoresis-mass spectrometry (CE-MS) analysis on M. leprae and compared to that of M. bovis BCG. Interestingly, comparison of these two profiles showed that, in M. leprae, amino acids and their derivatives are significantly accumulated, but most of intermediates related to central carbon metabolism markedly decreased, implying that M. leprae possess unique metabolic features. The present study is the first report demonstrating the unique profiles of M. leprae metabolites and these insights might contribute to understanding undefined metabolism of M. leprae as well as pathogenic characteristics related to the manifestation of the disease., Author Summary Mycobacterium leprae, the causative agent of leprosy, has unique physiological features including being uncultivable in artificial media. This fact raises the possibility that M. leprae possesses specific metabolism that are different from other cultivable mycobacteria. Among the components of M. leprae, the glycolipids are known to be involved in pathogenicity, while the dynamics of intracellular metabolites such as organic acids, amino acids and nucleic acids remain unclear. Aiming to understand the metabolism of M. leprae, we characterized the profile of intracellular metabolites. Unexpectedly, we found that amino acid species are significantly accumulated, while most of intermediates related to central carbon metabolism markedly decreased in the metabolite fraction of M. leprae, as compared with that of other mycobacteria. These specific metabolic features of M. leprae was presented for the first time and these insights may contribute to understanding the mechanism of physiology including obligate growth in vivo, which is one of the key characteristics of leprosy.

Paresthesia-dominant and pain-dominant subgroups have been noted in carpal tunnel syndrome (CTS), a peripheral neuropathic disorder characterized by altered primary somatosensory/motor (S1/M1) physiology. We aimed to investigate whether brain morphometry dissociates these subgroups. Subjects with CTS were evaluated with nerve conduction studies, whereas symptom severity ratings were used to allocate subjects into paresthesia-dominant (CTS-paresthesia), pain-dominant (CTS-pain), and pain/paresthesia nondominant (not included in further analysis) subgroups. Structural brain magnetic resonance imaging data were acquired at 3T using a multiecho MPRAGE T1-weighted pulse sequence, and gray matter cortical thickness was calculated across the entire brain using validated, automated methods. CTS-paresthesia subjects demonstrated reduced median sensory nerve conduction velocity (P = 0.05) compared with CTS-pain subjects. In addition, cortical thickness in precentral and postcentral gyri (S1/M1 hand area) contralateral to the more affected hand was significantly reduced in CTS-paresthesia subgroup compared with CTS-pain subgroup. Moreover, in CTS-paresthesia subjects, precentral cortical thickness was negatively correlated with paresthesia severity (r(34) = -0.40, P = 0.016) and positively correlated with median nerve sensory velocity (r(36) = 0.51, P = 0.001), but not with pain severity. Conversely, in CTS-pain subjects, contralesional S1 (r(9) = 0.62, P = 0.042) and M1 (r(9) = 0.61, P = 0.046) cortical thickness were correlated with pain severity, but not median nerve velocity or paresthesia severity. This double dissociation in somatotopically specific S1/M1 areas suggests a neuroanatomical substrate for symptom-based CTS subgroups. Such fine-grained subgrouping of CTS may lead to improved personalized therapeutic approaches, based on superior characterization of the linkage between peripheral and central neuroplasticity.

Autonomic nervous system (ANS) response to acupuncture has been investigated by multiple studies; however, the brain circuitry underlying this response is not well understood. We applied event-related fMRI (er-fMRI) in conjunction with ANS recording (heart rate, HR; skin conductance response, SCR). Brief manual acupuncture stimuli were delivered at acupoints ST36 and SP9, while sham stimuli were delivered at control location, SH1. Acupuncture produced activation in S2, insula, and mid-cingulate cortex, and deactivation in default mode network (DMN) areas. On average, HR deceleration (HR-) and SCR were noted following both real and sham acupuncture, though magnitude of response was greater following real acupuncture and inter-subject magnitude of response correlated with evoked sensation intensity. Acupuncture events with strong SCR also produced greater anterior insula activation than without SCR. Moreover, acupuncture at SP9, which produced greater SCR, also produced stronger sharp pain sensation, and greater anterior insula activation. Conversely, acupuncture-induced HR- was associated with greater DMN deactivation. Between-event correlation demonstrated that this association was strongest for ST36, which also produced more robust HR-. In fact, DMN deactivation was significantly more pronounced across acupuncture stimuli producing HR-, versus those events characterized by acceleration (HR+). Thus, differential brain response underlying acupuncture stimuli may be related to differential autonomic outflows and may result from heterogeneity in evoked sensations. Our er-fMRI approach suggests that ANS response to acupuncture, consistent with previously characterized orienting and startle/defense responses, arises from activity within distinct subregions of the more general brain circuitry responding to acupuncture stimuli.

Pentraxin 3 (PTX3) is an acute-phase reactant that is involved in amplification of the inflammatory response and innate immunity. In the present study, we evaluated the relationship between PTX3 and serum amyloid A (SAA), another acute-phase reactant, in rheumatoid synoviocytes.PTX3 mRNA expression was examined by reverse transcription polymerase chain reaction, and PTX3 protein was measured by enzyme-linked immunosorbent assay.SAA induced PTX3 mRNA and PTX3 protein expression in rheumatoid synoviocytes. SAA-induced PTX3 expression was attenuated when rheumatoid synoviocytes were nucleofected with N-formyl peptide receptor ligand-1 (FPRL-1)-specific siRNA, suggesting the involvement of FPRL-1. Furthermore, SAA-induced PTX3 expression was inhibited by NF-κB or mitogen-activated protein kinase-specific inhibitors. Neither soluble TNF receptor (etanercept) nor recombinant IL-1 receptor antagonist affected PTX3 production by SAA-stimulated synoviocytes, suggesting that SAA directly induces PTX3.Our data suggest that SAA plays a role in the proinflammatory and immune responses in rheumatoid synovium by inducing PTX3. We provide the first evidence that the acute-phase reactant SAA, which is produced systemically by hepatocytes, perpetuates the rheumatoid inflammatory processes by inducing another proinflammatory molecule, PTX3, locally in rheumatoid synovial tissues.

Objective.Inhibition of intracellular signal transduction is considered to be a therapeutic target for chronic inflammation. The new Janus kinase (JAK)3 inhibitor CP690,550 has shown efficacy in the treatment of rheumatoid arthritis (RA). We investigated the influence of JAK/STAT inhibition using CP690,550 on the induction of acute-phase serum amyloid A (SAA), which is triggered by interleukin 6 (IL-6) stimulation in rheumatoid fibroblast-like synoviocytes (RA-FLS).Methods.IL-6-stimulated gene expression of the acute-phase serum amyloid A genes (A-SAA; encoded by SAA1+SAA2) and SAA4 was analyzed by reverse transcriptase-polymerase chain reaction. The intracellular signaling pathway mediating the effects of CP690,550 on IL-6-stimulated JAK/STAT activation was assessed by measuring the phosphorylation levels using Western blots.Results.IL-6 trans-signaling induced A-SAA messenger RNA (mRNA) expression in RA-FLS. By contrast IL-6 stimulation did not affect SAA4 mRNA expression, which is expressed constitutively in RA-FLS. IL-6 stimulation elicited rapid phosphorylation of JAK2 and STAT3, which was blunted by CP690,550. CP690,550 abrogated IL-6-mediated A-SAA mRNA expression in RA-FLS. Similarly, CP690,550 inhibited IL-6-mediated A-SAA mRNA expression in human hepatocytes.Conclusion.Our data indicated that CP690,550 blocked IL-6-induced JAK2/STAT3 activation, as well as the induction of A-SAA. Inhibition of IL-6-mediated proinflammatory signaling pathways by CP690,550 may represent a new antiinflammatory therapeutic strategy for RA and AA amyloidosis.

Previously, we observed that both major membrane protein II of Mycobacterium leprae (MMP-ML) and its fusion with M. bovis BCG (BCG)-derived heat shock protein 70 (HSP70) (Fusion-ML) are immunogenic and that recombinant BCG secreting either of these proteins effectively inhibits the multiplication of M. leprae in mice. Here, we purified M. tuberculosis -derived major membrane protein II (MMP-MTB) and its fusion with HSP70 (Fusion-MTB) in a lipopolysaccharide-free condition and evaluated their immunostimulatory abilities. Both MMP-MTB and Fusion-MTB activated monocyte-derived dendritic cells (DC) in terms of phenotype and interleukin-12 (IL-12) production, but Fusion-MTB more efficiently activated them than MMP-MTB did. The IL-12 production was a consequence of the ligation of those recombinant proteins with Toll-like receptor 2. The M. tuberculosis -derived and M. leprae -derived recombinant proteins activated naïve T cells of both CD4 and CD8 subsets, but M. tuberculosis -derived proteins were superior to M. leprae -derived proteins and fusion proteins were superior to MMP, regardless of the origin of the protein. Memory-type CD4 + T cells obtained from BCG-vaccinated healthy individuals seem to be primed with MMP-MTB by the vaccination, and both M. tuberculosis -derived recombinant proteins produced perforin-producing CD8 + T cells from memory-type CD8 + T cells. Further, infection of DC and macrophages with M. tuberculosis H37Ra and H37Rv induced the expression of MMP on their surface. These results indicate that M. tuberculosis -derived MMP, as a sole protein or as part of a fusion protein, may be useful for developing new vaccinating agents against tuberculosis.