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Conventional diagnostic methods for tuberculosis (TB) have limited sensitivity and specificity or are time-consuming.16S rDNA and 16S rRNA of Mycobacterium tuberculosis complex (MTC) were used as targets to develop sensitive and specific polymerase chain reactions (PCRs) to improve the diagnosis of MTC.We developed conventional and real-time PCRs targeting 16S rDNA and rRNA of MTC.PCRs targeting 16S rRNA had a 10-100 times lower limit of detection for M. tuberculosis than PCRs targeting 16S rDNA. The sensitivities of the 16S rDNA PCR, 16S rRNA reverse transcription PCR (RT-PCR), 16S rDNA real-time PCR and 16S rRNA real-time RT-PCR for sputum specimens were respectively 92%, 94.6%, 96% and 100%. Real-time PCR showed no cross-reactivity, but conventional PCR had cross-reactivity to M. avium, M. gastri and M. nonchromogenicum.PCRs targeting the 16S rRNA of MTC were more sensitive than those targeting 16S rDNA; 16S rRNA real-time RT-PCR showed the highest sensitivity and specificity for MTC.

Leakage current characteristics have been investigated for nitride-based InGaN light-emitting diodes (LEDs) with different levels of Si-doping conditions applied to the n-type InGaN/GaN layer. The tested LED samples had an emission wavelength of 445 nm. Based on the analysis of temperature-dependent characteristics measurements, the reverse leakage current, IR, was predominantly influenced by the lowering of Poole-Frenkel (PF) barrier for low electric field ( ;8 × 107 V/cm), phonon-assisted tunneling (PAT) dominates the IR mechanism. The IR is influenced by the Si-doping density of InGaN/GaN layer that may be present near the end of depletion regions of tested samples.

Dynamical systems may be either discrete or continuous, though it is often convenient to blur this distinction in order to formulate a continuum model of a large discrete system. For example, in systems in which macroscopic behavior alone is of interest, a continuum approach is justified because of the vast number of molecules present. But just how many molecules are necessary for a system to be accorded a continuum description? Due to recent advances in interrogating, fabricating, and controlling small collections of molecules [1], it now seems appropriate to consider the manner in which discrete physical systems acquire a continuum description. Such a study is not limited, of course, to molecular systems. As a preliminary question, we may ask, “How can the terms ‘small’ and ‘large’ be made precise?” To formulate this question in a physical context, we consider a model dynamical problem originally investigated by Fermi, Pasta, and Ulam (FPU) [2]. The model consists of a one-dimensional chain of point masses that interact with their nearest neighbors via nonlinear forces. FPU were surprised to find that even in the presence of nonlinearity the dynamics of the system was almost periodic, with a period known as the recurrence time. Further investigations of the lattice dynamics led to the discovery that the recurrence time has a power-law dependence on system parameters [3] (see also [4,5]). This can be explained in the continuum limit by soliton solutions of the Korteweg-de Vries (KdV) equation [6–9]. In this model, derived by letting the number of lattice points tend to infinity while keeping the total length of the lattice fixed [10], solitons flow around the lattice independently; the recurrences are due to the periodic return of these solitons to the initial configuration. The well-established scaling of recurrence time in the continuum limit makes the FPU lattice an ideal system to study the transition from a discrete lattice to a continuum system. We focus here on the behavior of the lattice as it passes through the twilight between “small” and “large” by examining the FPU recurrences as the system size changes. For large systems, when the dynamics is well modeled by the continuum KdV equation, we find that the recurrence can be characterized by a single parameter that measures the strength of the nonlinearity of the system. As the system size shrinks, however, a second parameter is required: the size of the system itself. The transition between these two regimes is well-defined and occurs at a critical value of the system size that appears to coincide with the onset of an instability in the dynamics. We are therefore able to formulate a precise criterion for where the transition between “small” and “large” lattices occurs. The FPU model consists of a one-dimensional lattice of point masses coupled by a linear force augmented by a quadratic nonlinearity and is described by the Hamiltonian

The Ni-dot/Ag/Pt layer, where Ni-dot layer is formed of nanometer sized Ni dots, has been used to improve the reflectivity from the surface of p-type GaN in a light emitting diode (LED). Comparing with Ni/Ag/Pt layer, where Ni layer is a thin film, the Ni-dot/Ag/Pt structure shows significantly improved reflectivity with stable contact resistivity. The optical output power and external quantum efficiency of InGaN LEDs with Ni-dot/Ag/Pt structure for p-metal have improved by 28% and 29%, respectively, over the results of Ni/Ag/Pt structure.

Extracellular newly identified RAGE-binding protein (EN-RAGE) is a ligand of the receptor for advanced glycation endproducts (RAGE) and has been termed S100A12. The ligation of EN-RAGE with RAGE on the endothelium, mononuclear phagocytes and lymphocytes triggers cellular activation with the generation of the key proinflammatory mediators interleukin (IL)-1beta and tumour necrosis factor (TNF)-alpha.The aim of this study was to investigate the presence of RAGE and EN-RAGE, their spatial localization and their coexpression in leprosy lesions.Immunohistochemistry and confocal laser scanning microscopy were used to evaluate the expression of RAGE and EN-RAGE in leprosy. By enzyme-linked immunosorbent assay, RAGE and EN-RAGE were detected in the serum.(1) In the multibacillary (MB) and paucibacillary (PB) groups, the level of RAGE production was significantly higher than in patients with atypical mycobacterial infection or sarcoidosis (P0.01). In the MB group, the production of RAGE was higher than in the PB group (P0.01), and it was higher in patients without the lepra reaction than in patients with the lepra reaction (P0.05). (2) In MB, PB and atypical mycobacterial infection, the level of EN-RAGE production was significantly higher than in sarcoidosis (P0.01). (3) In the confocal laser scanning microscopic examination, the RAGE and EN-RAGE proteins were detected in lepromatous leprosy. These proteins are spatially colocalized along the cell surface, which is in agreement with their receptor-ligand interaction. (4) A comparable amount of EN-RAGE was detected in the serum of the MB and PB groups. Patients with the reaction showed a higher level of EN-RAGE than patients without the reaction in leprosy.Our data suggest that in leprosy, RAGE and EN-RAGE may be involved in the proinflammatory process rather than the antimycobacterial activity, especially during the lepra reaction. The blockade of the interaction of RAGE and EN-RAGE at the early stage of the inflammatory process may minimize the inflammatory response and consequent tissue damage or the sequelae of leprosy.

The single base change at the 94th codon of inhA has been referred to as the event that confers resistance on the drugs isoniazid (INH) and ethionamide (ETH) in Mycobacterium smegmatis and M. bovis. From this observation, it has been anticipated that some of the INH-resistant clinical isolates of M. tuberculosis would carry missense mutations in the same region of the gene. However, few polymorphisms have been identified in this region among INH-resistant isolates.To understand the molecular basis for M. tuberculosis resistance to INH and ETH.The sequence polymorphism at the 94th codon of inhA among M. tuberculosis isolates from Korea was analyzed by polymerase chain reaction (PCR) cloning and sequence analysis.No nucleotide change at the 94th codon of inhA was detected in any of the 24 INH-resistant isolates analyzed in this study. On the other hand, a point mutation was found exclusively at the regulatory region flanking a putative ribosome-binding site of the inhA locus in 14 isolates. Interestingly, all the mutations were of the same kind, which substitutes C to T. Among 14 isolates, 12 were resistant to INH as well as to ETH, while two were resistant to INH only.It seems that mutations previously found at the 94th codon of inhA have no particular relationship with the mechanism involved in the resistance of M. tuberculosis to INH and/or ETH. On the other hand, the resistance mechanism of M. tuberculosis to INH/ETH may involve an altered level of InhA, an expression which may have been influenced by the sequence change in the regulatory region of the inhA locus.

Agglutinating antibodies to neuraminidase-treated red blood cells (anti-T agglutinins) are known to be reduced in patients with gastric cancer. The antigenic determinant of anti-T agglutinin is known to have a disaccharide structure [Gal(beta1-3)GalNAc], the same specificity as peanut agglutinin (PNA). We examined sera of 27 patients with gastric cancer and 30 controls for anti-T agglutinins, anti-T antibodies and PNA-binding glycoproteins. Anti-T agglutinins were titrated by a microtiter hemagglutination method. Levels of anti-T antibodies were determined by enzyme immunoassay using synthetic glycoconjugate [Gal(beta1-3)GalNAc O-alpha-linked to human serum albumin] as an antigen. Levels of PNA-binding glycoproteins in sera were measured by sandwich enzyme-linked lectin assay using wheat germ agglutinin and peroxidase-conjugated PNA. Titers of anti-T agglutinins were significantly lower in patients with gastric cancer than in controls (P = 0.041). Levels of anti-T antibodies were not significantly different in patients with gastric cancer and controls; however, decreased levels of anti-T antibodies were more frequent in patients with gastric cancer than in controls (P = 0. 001). Levels of PNA-binding glycoproteins were significantly higher in sera of patients with gastric cancer than in controls (P = 0.001). The levels of anti-T antibodies inversely correlated with the levels of PNA-binding glycoproteins in sera of patients with gastric cancer (r = -0.44, P = 0.021). These results suggest that the decrease in anti-T antibodies in sera of patients with gastric cancer might be due to immune complex formation between circulating PNA-binding glycoproteins and anti-T antibodies.

To assess the molecular mechanism of rifampin (RMP) resistance in clinical strains of Mycobacterium tuberculosis.The molecular nature of a part of the rpoB gene in 77 M. tuberculosis clinical strains isolated in Korea was analyzed using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and PCR-sequence analysis.Among 67 RMP-resistant isolates, 50 showed SSCP profiles different from that of an RMP-sensitive control strain, M. tuberculosis H37Rv, indicating the possible existence of a sequence alteration in this region of the rpoB gene, while 17 resistant isolates displayed SSCP profiles indistinguishable from that of the sensitive control strain. Subsequently, 17 clinical isolates whose SSCP profiles were difficult to distinguish from the control strain were subjected to sequence analysis. The analysis revealed that all 17 isolates did indeed contain mutations in the 81 bp region of the rpoB gene, which is associated with RMP resistance.The results from our study clearly indicate that the molecular mechanism of RMP resistance in M. tuberculosis isolates from Korea involves alterations in the rpoB gene. In addition, this study suggests that PCR-direct sequence analysis works more efficiently and accurately than PCR-SSCP analysis for rapid screening of RMP-resistant M. tuberculosis clinical isolates.

Using sera from 4 pairs of mangabey monkeys inoculated with titrated doses of Mycobacterium leprae we demonstrated that IgA antibodies against M. leprae specific PGL-I antigen were present in 75% of inoculated monkey's sera. High IgA antibody was detected in 50% (3/6) of infected animals and all three developed lepromatous leprosy (LL). Antibody titers correlated with PGL-I antigen in serum. The highest IgA peak appeared late and corresponded to the beginning of treatment, and in two of them appeared shortly after or corresponded with neurological damage. Low IgA response was found in the other 3 monkeys (50%-3/6), two of which developed indeterminate leprosy (I) and the other one LL. Low IgA levels appeared late after IgG and IgM, and shortly after neurologic signs. Both I monkeys were negative for PGL-I in serum. The remaining 2 monkeys (25%-2/8) did not show an IgA response; one of them developed LL but the disease regressed to I. IgM seemed to correspond to the appearance of PGL-I in serum. The other animal did not develop clinical symptoms of leprosy, and PGL-I in serum was negative. Although there was no clear relation between the development of anti-PGL-I IgA and experimental leprosy, the finding of a high IgA response in some animals suggests that further studies are needed to evaluate the role of antigen-specific IgA in the disease process.

Local synthesis of immunoglobulin G (IgG) in the central nervous system was investigated in 10 patients with tuberculous meningitis (TBM), 15 patients with aseptic meningitis (AM), and 15 patients with pulmonary tuberculosis only (PTBO). The IgG synthesis rate for patients with TBM was 56.4 +/- 18.9 mg/day (mean +/- standard deviation), which was significantly higher than that for patients with AM (8.0 +/- 6.7 mg/day, P < 0.001) and that for patients with PTBO (7.5 +/- 4.4 mg/day, P < 0.001). Therefore, the increased IgG synthesis rate in the central nervous system provided supporting evidence for differentiating the diagnosis of TBM from that of AM (sensitivity, 100%; specificity, 83.3%). Simultaneous measurement by enzyme-linked immunosorbent assay of IgG seroreactivity to lipoarabinomannan and purified protein derivative antigens in cerebrospinal fluid (CSF) demonstrated seropositivity in all 6 patients with TBM, 4 of 15 patients with AM, and 4 of 10 patients with PBTO. All patients showing false-positive reactivity in CSF demonstrated seropositivity in sera and normal ranges for IgG synthesis rates in CSF. Also, the semiquantitive measurement of IgG antibody (Ab) titers in these patients demonstrated higher IgG Ab titers in serum than in CSF except for one patient with a highly elevated albumin quotient, suggesting a leaky blood-brain barrier. The results strongly suggested that the Mycobacterium tuberculosis-specific IgG Abs were diffusible through the blood-brain barrier, which addresses the pitfall of serological tests for the early diagnosis of TBM. The serological detection of IgG Abs to lipoarabinomannan and purified protein derivative antigens in CSF could be misleading in the presence of simultaneously elevated of IgG Abs in serum.

Eight sooty mangabey monkeys were inoculated intravenously and intradermally with varying doses of Mycobacterium leprae from 4.8 x 10(7) to 4.8 x 10(10). Serum samples were obtained from the animals at intervals of about 3 months for 90 months, and were examined for IgM and IgG antibodies to nerve antigens, including ceramide, galactocerebroside (GC), and asialo-GM1 (AGM1), using an enzyme-linked immunosorbent assay (ELISA). The serological results were then compared with clinical findings, particularly nerve involvement. Of 8 mangabey monkeys inoculated with M. leprae, 7 animals had clinical leprosy; 6 of them had nerve damage, including neurologic deformities in 4 monkeys and nerve enlargement in 2. Median time for the initial signs of leprosy was 10 months postinoculation (p.i.), a range from 4 to 35 months. In contrast, nerve damage was noted rather late, about 35 to 86 months p.i. (median 54 months). The major immunoglobulin class to ceramide, GC, and AGM1 antigens was IgM, and the antibody responses to the nerve antigens appeared from 15 to 63 months p.i. (median 37 months). Antineural antibodies were thus detectable about 18 months (range -2 to 60 months) prior to observable nerve damage. In addition, elevation of antineural antibody levels were predictive of clinical exacerbation of the disease and neuritic damage. This study suggests that antineural antibodies are produced during the course of M. leprae infection and may be indicative of nerve damage, such as neurological deformities or nerve enlargement, in leprosy patients.

P-type AlGaN/GaN/InGaN superlattices were incorporated in a InGaN based blue light emitting diode as electron blocking layer to minimize the temperature dependence on optical output power. For the characteristic temperatures in range of 10 to 100 °C and at operation current of 350 mA, the external quantum efficiency varied by less than 0.5%. For the presented device, the negative characteristic temperature was shown to occur below temperature of 50 °C. The improved temperature stability in optical output power is thought to be attributed to (1) the efficiency of hole carrier transport in AlGaN/GaN/InGaN superlattices and (2) the enhanced blocking of electron overflow between multiple quantum wells and AlGaN/GaN/InGaN superlattices.

Since phenolic glycolipid-I (PGL-I) is an unequivocal marker of Mycobacterium leprae, the antigen has been a good candidate for the serodiagnosis and monitoring the effectiveness of leprosy chemotherapy. As an effort to define the kinetics of the PGL-I antigen and its antibodies in leprosy patients, this study was initiated to examine the serum specimens obtained serially from lepromatous patients under chemotherapy trials. PGL-I was detectable in 64 (94.1%) of 68 new lepromatous (bacterial index, BI = 3.2 to 5.8) and in 26 (78.8%) of 33 relapsed lepromatous patients (BI = 3.0 to 5.3). Meanwhile, virtually all of the new and relapsed patients were strongly seropositive to PGL-I. PGL-I was not detectable in any of the patients about 18 months after chemotherapy was initiated; however, anti-PGL-I reactivity declined by 50% at 2 years and by about 70% at 5 years after chemotherapy regardless of the drug regimens under study. Considering the rapid disappearance of the PGL-I antigen and steady decrease in anti-PGL-I IgM antibodies following chemotherapy, the PGL-I-based serology may be useful for monitoring the effectiveness of treatment, at both the early and late stages, in leprosy patients whose initial sera contain a significant level of PGL-I antigen or antibodies.

The current approaches in serological methods for the diagnosis of tuberculosis have been towards the use of M. tuberculosis-specific antigens or epitopes of protein antigens. Although sensitivity by using such antigens might be similar to that obtained by using crude or semipurified antigens, specificity was somewhat improved to over 95%. Among serological tools, ELISA has been the method of choice for detecting serum antibodies and mycobacterial antigens from clinical specimens because of its high sensitivity, simplicity, reproducibility and versatility in screening a large number of specimens. Further simple and rapid tests such as the agglutination test by coating various particles with purified or synthetic M. tuberculosis-specific antigens or antibodies will soon become available for field evaluation in endemic areas of tuberculosis. However, all these efforts can be justified only if serological tests give any meaningful information for the diagnosis and management of tuberculosis or for determining the degree of M. tuberculosis transmission in a community as a complement to the current tuberculin test, chest X-ray, or sputum examination. Although the detection of mycobacterial antigens from CSF or sputum specimens using polyclonal or monoclonal antibodies gave more direct evidence for M. tuberculosis infection, the information by the assays was not sufficient to determine whether these immunological tools could be used in clinical laboratories for the diagnosis of tuberculosis. Meanwhile, recent introduction of the very sensitive polymerase chain reaction (PCR) technology for the detection of M. tuberculosis DNA from clinical specimens (37-40) may eventually replace the immunological assays for the detection of mycobacterial antigens.(ABSTRACT TRUNCATED AT 250 WORDS)

Sequential monitoring of 724 sera for antibodies to a neoantigen based on phenolic glycolipid-I (PGL-I) and native lipoarabinomannan (LAM) in 90 leprosy patients undergoing therapy in San Francisco was conducted. Untreated lepromatous patients frequently (91%) had significant antibodies to both moieties. Antibodies were less frequently found in tuberculoid patients (74% to neoantigen and 37% to LAM). In the first 3 years of treatment, average serum antibodies to both moieties fell significantly. Antibodies to LAM fell during each of the first 4 years of therapy, but decreasing antibody levels to the PGL-I neoantigen did not appear to fall consistently after the third year of treatment. A wide variation in the rate of fall of serum antibodies was noted. Sequential changes in the amounts of serum antibodies to the neoantigen and LAM in general paralleled one another but were at times discrepant. Both in San Francisco and Malaysia, skin-smear negative, long-term treated, lepromatous leprosy patients frequently harbored significant antibodies to both PGL-I and LAM.