Molecular screening of multidrug-resistance tuberculosis by a designated public health laboratory in Taiwan.

This manuscript describes our experience in early identifying MDR-TB cases in high-risk populations by setting up a single-referral molecular diagnosis laboratory in Taiwan. Taiwan Centers for Disease Control designated a single-referral laboratory to provide the GenoType MTBDRplus test for screening high-risk MDR-TB populations nationwide in 2012-2015. A total of 5,838 sputum specimens from 3,308 patients were tested within 3 days turnaround time. Compared with the conventional culture and drug susceptibility testing, the overall performance of the GenoType MTBDRplus test for detecting TB infection showed accuracy of 70.7%, sensitivity of 85.9%, specificity of 65.7%, positive predictive value of 45.5%, and negative predictive value of 93.3%. And the accuracy of detecting rifampin (RIF) resistance, isoniazid (INH) resistance, and MDR-TB (resistant to at least RIF and INH) were 96.5%, 95.2%, and 97.7%, respectively. MDR-TB contacts presented a higher rate of mutated codons 513-519, GenoType MTBDRplus banding pattern: rpoB WT3(-), and rpoB WT4(-) than the treatment failure group. The MDR-TB contact group also had a higher rate of inhA C15T mutation, banding pattern: inhA WT1(-), and inhA MUT1(+) than the recurrent group. Resistance profiles of MDR-TB isolates also varied geographically. The referral molecular diagnosis system contributed to rapid detection and initiation of appropriate therapy.

Effect of high glucose, Porphyromonas gingivalis lipopolysaccharide and advanced glycation end-products on production of interleukin-6/-8 by gingival fibroblasts.

BACKGROUND AND OBJECTIVE: It is known that chronic periodontal infection can magnify the cytokine responses in patients with diabetes. Hyperglycemia increases the proinflammatory status, including the levels of advanced glycation end-products (AGEs), in patients with periodontitis. However, whether AGEs have additional effects on the production of those proinflammatory cytokines in diabetic patients with periodontitis is still unknown. To examine in vitro the effect of hyperglycemia and AGEs on the amounts of interleukin (IL)-6 and IL-8 produced in periodontally infected gingiva, human gingival fibroblasts (HGFs) were stimulated with glucose, AGE-modified bovine serum albumin (AGE-BSA) and Porphyromonas gingivalis LPS in the present study. MATERIAL AND METHODS: Primary culture of HGFs was incubated with various concentrations of AGE-BSA (0, 50, 100 and 200 µg/mL) and LPS (0, 10, 100 or 1000 ng/mL) at two different glucose concentrations - normal glucose (5 mm) and high glucose (25 mm). The amounts of IL-6 and IL-8 produced by HGFs were evaluated using ELISA. Expression of the AGE receptor on HGFs was determined by flow cytometry. RESULTS: High glucose stimulated a significant increase in the production of IL-6 and IL-8 by HGFs compared with normal glucose. This enhanced production of IL-6 and IL-8 could also be observed in the presence of LPS and/or AGE-BSA. When both LPS and AGE-BSA were present, especially at high concentrations (≥ 500 µg/mL of LPS and ≥ 25 µg/mL of AGE-BSA), a synergistic effect on IL-8 production was found in the high-glucose condition. CONCLUSIONS: A synergistic effect of the production of IL-8 could be induced in HGFs with the combination of high glucose, LPS and AGEs.

A possible case of hantavirus infection in a Borneo orangutan and its conservation implication.

BACKGROUND: Natural infection of hantavirus in orangutans has never been reported. METHODS: Enzyme-linked immunosorbent assays (ELISA), Indirect immunofluorescence assay (IFA), and RT-PCR were used to diagnosis a suspected case in a pet orangutan in southern Taiwan. RESULTS: Although the RT-PCR result was negative, the high IgG titer in the beginning and its dramatic drop after treatments suggested a recent Seoul-type hantavirus infection. CONCLUSIONS: Hantavirus transmission and its potential damage to wild orangutans should not be overlooked.

Analysis of bcl-2 expression in normal, inflamed, dysplastic nasopharyngeal epithelia, and nasopharyngeal carcinoma: association with p53 expression.

To further characterize bcl-2 expression in nasopharyngeal carcinoma (NPC), the authors analyzed bcl-2 expression immunohistochemically in biopsy specimens from 101 cases of NPC, of which 65 had the component of normal nasopharyngeal epithelium (NPE), 24 with dysplastic lesions adjacent to carcinoma, and 14 with both primary and metastatic lesions. An additional 25 nasopharyngeal biopsies of NPE from patients with chronic inflammation of nasopharynx were also included. The percentage of detectable bcl-2 expression shown in NPC (80%) and adjacent dysplastic lesions (71%) was significantly higher than in adjacent NPE (37%) and NPE from patients with chronic inflammation of the nasopharynx (30%) (P < .05). In both normal and inflamed NPE, the detectable bcl-2 expression was restricted to the basal cells; however, in dysplastic lesions, the bcl-2 staining distribution was increased with the dysplastic cell layers, and in entire layers of epithelium in severe dysplasia or carcinoma in situ. In addition, the staining intensity of bcl-2 in carcinomas and adjacent dysplastic lesions was generally stronger than that of adjacent NPE. These observations suggest that the expression of bcl-2 in dysplasia and carcinoma is enhanced relative to that of adjacent NPE. Enhanced bcl-2 expression to prevent apoptosis seems to occur from the early stages and may play an important role in the carcinogenesis of NPC. Furthermore, up to 77% of NPC with the coexpression of bcl-2 and p53 was observed and suggested that the association of bcl-2 and p53 expression seems to occur from the early stages of the development of NPC. The overexpression of p53 protein in NPC suggests that the mutation of p53 gene or altered function of wild-type p53 protein may contribute to the pathogenesis. It is conceivable that the presence of both enhanced bcl-2 expression and altered p53 functions may play a crucial synergistic effect in the carcinogenesis of NPC.

Effect of positioning on pulmonary function of newborns: comparison of supine and prone position.

The effect of positioning on pulmonary function has been previously evaluated, and the prone position has been reported to be preferable for neonates with various respiratory diseases. Studies in healthy neonates have yielded conflicting results. Using a crying pulmonary function test, we examined the effect of positioning on pulmonary function in healthy full-term neonates. Thirty-nine infants with a mean birthweight (+/- SD) of 3,140 +/- 379 g and a mean gestational age (+/- SD) of 39.8 +/- 1.6 weeks were investigated during the first 6 hours of life. Measurements were obtained in both supine and prone positions using a computerized volume-flow system. There were statistically significant decreases in crying vital capacity (CVC) and peak expiratory flow rate (PEF) in the prone compared with the supine position. However, there were no significant differences in forced expiratory flow rate at 75% (V(75)), 50% (V(55)), and 25% (V(25)) of vital capacity between the two positions. These results suggest that prone positioning decreases lung volume and increases resistance of upper airways. We conclude that healthy neonates should be in the supine posture for optimal ventilation.

Gingival overgrowth and dental alveolar alterations: possible mechanisms of cyclosporin-induced tooth migration. An experimental study in the rat.

The inter-incisal distance and dimension of the interdental papilla between the mandibular incisors were examined in cyclosporin A (CSA) fed rats over 6 weeks. Rats in the test group received CSA daily in mineral oil by gastric feeding (30 mg/kg body weight); the control group received mineral oil only. The inter-incisal distance and gingival dimensions, including bucco-lingual width and vertical height, were assessed biweekly from alginate impressed stone models. Animals were sacrificed at the end of the study and tissue sections were obtained from the anterior region of the mandible for histopathological evaluation. Both the inter-incisal distance and the dimension of the interdental papilla were significantly greater in CSA-exposed animals compared to control. The significant alteration appeared earlier in the papillary dimensions than that in the interdental distance. Particular histopathological alterations of the soft and hard tissues of the periodontium were observed in CSA-exposed animals. Within limitations of the study, we suggest that the CSA-induced gingival overgrowth may offer an active force contributing to observed tooth movement, however, remarkable alveolar remodeling should be considered as an undetermined factor for the movement.

[Trial vaginal delivery for women with previous cesarean section].

A retrospective analysis was made on 85 cases of trial labor after previous cesarean section (CS) and compared with 100 primiparous vaginal deliveries (as control) during the last 10 years. Oxytocin infusion has been used in 61 trial cases to induce and/or to accelerate labor, and valium, procaine and atropine administered during the labor process in 23 of them. 65 of the 85 cases (76.5%) successfully delivered through the vagina (VD group), while 20 cases (23.5%) had repeat CS following failure in trial labor (CS group), and 3 women had reatened uterine rupture. There was 1 neonatal death in the VD trial labor group. No obvious difference in the Apgar scores of newborns was found between the CS group and the controls, and the duration of the labor process was much shorter in the VD trial group than that in the primiparae (7.51 +/- 2.44 hrs vs. 9.10 +/- 3.75 hrs) (P < 0.01). This study indicated that following a previous uneventful transverse lower segment cesarean section, trial labor should be encouraged under supervision.

[Outcome of gestation of 32 pregnant women with toxoplasma and cytomegalovirus infection].

In an attempt to investigate the harmful effects of the infection of Toxoplasma and associated with cytomegalovirus on the fetus infection acquired transplacentally during their mothers' pregnancy sera from 1000 pregnant women were tested for antibodies to both pathogens and the circulating antigen of Toxoplasma by using ELISA. The test was positive (titre > or = 1:60400) in 32 of 1000 cases. Judging from the sero-positive results together with the case history, clinical manifestations and B-scan results, the 32 cases were advised to end their pregnancy. Necropsy and pathological examinations of the foetuses revealed stillborn foetus, hydrocephalus, acephalus, pleural effusion, ascites and other congenital malformations. Toxoplasma gondii were found in the tissues of 21 fetuses.

A role for maternal physiological state in preserving auditory cortical plasticity for salient infant calls.

A growing interest in sensory system plasticity in the natural context of motherhood has created the need to investigate how intrinsic physiological state (e.g., hormonal, motivational, etc.) interacts with sensory experience to drive adaptive cortical plasticity for behaviorally relevant stimuli. Using a maternal mouse model of auditory cortical inhibitory plasticity for ultrasonic pup calls, we examined the role of pup care versus maternal physiological state in the long-term retention of this plasticity. Very recent experience caring for pups by Early Cocarers, which are virgins, produced stronger call-evoked lateral-band inhibition in auditory cortex. However, this plasticity was absent when measured post-weaning in Cocarers, even though it was present at the same time point in Mothers, whose pup experience occurred under a maternal physiological state. A two-alternative choice phonotaxis task revealed that the same animal groups (Early Cocarers and Mothers) demonstrating stronger lateral-band inhibition also preferred pup calls over a neutral sound, a correlation consistent with the hypothesis that this inhibitory mechanism may play a mnemonic role and is engaged to process sounds that are particularly salient. Our electrophysiological data hint at a possible mechanism through which the maternal physiological state may act to preserve the cortical plasticity: selectively suppressing detrimental spontaneous activity in neurons that are responsive to calls, an effect observed only in Mothers. Taken together, the maternal physiological state during the care of pups may help maintain the memory trace of behaviorally salient infant cues within core auditory cortex, potentially ensuring a more rapid induction of future maternal behavior.

Alterations of gingival morphology in nifedipine-fed rats.

BACKGROUND: Gingival enlargement induced by nifedipine (NIF), a calcium antagonist, has been reported in human and animal studies. However, three-dimensional morphologic measurements of gingivae have never been used to describe this type of enlargement. We previously established an animal model of cyclosporin-induced gingival enlargement. The present study used morphologic measurements to examine whether or not NIF-induced gingival enlargement occurs in the animal model. METHODS: Eighty male Sprague-Dawley rats were divided into four groups. Rats in each group received daily NIF, at a dosage of 0, 10, 30 or 50 mg/kg body weight, by gastric feeding for nine weeks. Gingival dimensions (including buccolingual and mesiodistal widths, and vertical height) were assessed from stone models obtained from the mandibular incisor regions every three weeks. RESULTS: Over the course of the nine-week experiments, significant dimensional changes of the gingivae were observed according to two main factors: 1) the dose or, 2) the treatment duration. Gingival dimensions (including buccolingual and mesiodistal widths, and vertical height) significantly increased with the duration of NIF treatment. Dimensional alterations of gingivae were noted among the different dosage groups, but significant differences were mainly observed in those groups compared to the 50 mg/kg group. CONCLUSIONS: The finding of increased gingival morphology in NIF-treated rats in this study shows that gingival enlargement can be induced by NIF in rats.

Evidence that polymorphism of the angiotensin I converting enzyme gene may be related to idiopathic dilated cardiomyopathy in the Chinese population.

Angiotensin I-converting enzyme (ACE) is responsible for the production of angiotension II and the breakdown of kinins, leading to increased blood pressure (BP), induction of vascular smooth muscle cell proliferation, and the stimulation of myocardial-cell hypertrophy. A 287 bp insertion/deletion polymorphism in intron 16 of the ACE gene was examined by polymerase chain reaction in a cross-sectional study of 35 patients with idiopathic dilated cardiomyopathy (IDC) and 35 patients with normally functioning hearts (NT). Compared with the deletion/deletion (D/D) frequency in the control population, the frequency of the deletion allele was 0.757 in IDC patients and 0.600 in NTs; the difference between observed alleles in all subjects in each group was significant (x2 = 3.96; P < 0.05). The data thus provide evidence in favor of an association between idiopathic dilated cardiomyopathy and a polymorphism at the ACE locus (17q23), thus implicating this locus, and possibly a genetic variant of ACE, itself, in human idiopathic dilated cardiomyopathy.

Downregulation of phospholipase C delta3 by cAMP and calcium.

Four different isoforms of mammalian phospholipase C delta (PLCdelta) have been described. PLCdelta1, the best-understood isoform, is activated by an atypical GTP-binding protein. It has been suggested that it is a calcium signal amplifier. However, very less is known about other subtypes, including PLCdelta3. Therefore, in the present study, we examined the expression of PLCdelta3 in different human tissues. Moreover, the cellular underlying regulation for PLCdelta3 was studied in different cell lines. Our study showed that the mRNA and protein levels differed significantly among human tissues. The human PLCdelta3 gene was composed of 15 exons and 1 putative cAMP response element in the 5'-end promoter region. PLCdelta3 mRNA expression was downregulated by cAMP and calcium in both the human normal embryonic lung tissue diploid WI38 cell line and the glioblastoma/astrocytoma U373 cell line. However, mRNA expression showed no impact by PKC activators or inhibitors. This study shows the human PLCdelta3 expression pattern and is the first report that PLCdelta3 gene expression is downregulation by cAMP and calcium.

Interleukin-1 receptor antagonist modulates the progression of a spontaneously occurring IgA nephropathy in mice.

Cytokines, such as interleukin-1 (IL-1), may play a key role in the pathogenesis of IgA nephropathy (IgAN). This study was conducted to evaluate the effects of IL-1 receptor antagonist (IL-1ra) in the treatment of a spontaneously occurring experimental IgAN in established phase. ddY mice (12/group) were injected twice daily with 3 mg/kg of IL-1ra, intraperitoneally, for 8 consecutive weeks. The placebo mice were injected with saline only. As normal controls, ddY mice, which were not treated with IL-1ra or saline, were killed at 6 weeks of age. Results showed a significant reduction of proteinuria in the IL-1ra-treated mice, compared with saline-treated mice (urinary albumin/creatinine, 0.24 +/- 0.04 v 0.39 +/- 0.03, P < 0.001). A significant improvement of renal 51Cr-EDTA (ethylenediaminetetra-acetic acid) clearance was observed in the IL-1ra-treated mice (t1/2, 12 +/- 2.7 minutes, compared with saline-treated mice 25 +/- 2.0 minutes, P < 0.001). Similarly, serum levels of creatinine (1.0 +/- 0.4 v 2.4 +/- 0.3 mg/dL, P < 0.001) and urea nitrogen (46 +/- 6 v 58 +/- 2 mg/dL, P < 0.01) were significantly lower in IL-1ra-treated mice than in saline-treated mice. In renal tissue studies, the IL-1ra-treated mice exhibited significantly decreased mesangial cell proliferation, compared with saline-treated mice (P < 0.001), as shown by light and electron microscopy. In addition, the IL-1ra-treated mice showed significantly lower glomerular expression of collagen type IV, fibronectin, laminin, and IL-6 (P < 0.001) than saline-treated mice, although they still showed higher glomerular expression of collagen type IV (P < 0.01), fibronectin (P < 0.01), laminin (P < 0.001), IL-1 (P < 0.001), and IL-6 (P < 0.01) than did normal control mice. Meanwhile, glomerular C3 deposition was significantly lower in IL-1ra-treated mice than in saline-treated mice (P < 0.001). These findings indicate that IL-1ra partially prevented the progression of spontaneously occurring IgAN in this experimental model. Data from these experiments also confirm the pathogenic effects of IL-1 in the established phase of IgAN in ddY mice.

Down regulation of bcl-2 by p53 in nasopharyngeal carcinoma and lack of detection of its specific t(14;18) chromosomal translocation in fixed tissues.

High levels of bcl-2 protein have been found in a wide variety of human cancers. Since p53 gene inactivation occurs in over half of human cancers, it is possible that loss of p53-mediated repression of bcl-2 gene expression accounts, at least in part, for the frequent abnormalities in bcl-2 protein production seen in tumours. By using immunohistochemical methods, we have analysed thirty-three nasopharyngeal carcinomas for p53 and bcl-2 expression. We found an inverse correlation between the expression of these two proteins (P < 0.001). Moreover, we utilized universal oligonucleotide primers of a region 5' to the bcl-2 MBR and at the 3' end of JH segments to initiate a DNA polymerase chain reaction that amplified these bcl-2-JH junctures. Of the twelve nasopharyngeal carcinomas expressing bcl-2, none showed a t(14;18) chromosome translocation. These findings may indicate potential mechanisms by which bcl-2 regulates apoptosis.

Involvement of immunopathogenic mechanisms in a spontaneously occurring glomerulopathy in mice.

Mice have been found to be susceptible to spontaneous renal localization of immune deposits. However, the significance of these immune deposits is still debated. We investigated the immunopathogenesis of a naturally occurring glomerulopathy associated with progressive proteinuria and glomerulosclerosis in 75 BALB/c mice. The mice were divided into five groups of 15 and killed at the age of 1, 3, 6, 12, or 18 months for laboratory and renal pathologic studies. These mice showed persistently increasing serum levels of immune complexes, a marked increase of glomerular immune deposits which were capable of fixing C3, and interstitial infiltration of lymphocytes and plasma cells, followed by the occurrence of proteinuria, mesangiopathy, and glomerulosclerosis. Our findings suggest that an immune system mediated process occurred in the kidneys of the mice tested.

Soluble CD44 isoforms in serum as potential markers of metastatic gastric carcinoma.

A splice variant of CD44 (exon V4-V7) confers metastatic behavior in a rat carcinoma model; aberrant expression of splice variants has been detected on a variety of human tumor cell lines as well as primary and metastatic human tumors, including lymphomas, carcinomas (colon, thyroid, mamma, bladder), and glioma. We used enzyme-linked immunosorbent assay to determine the concentration of soluble CD44 in the serum samples of 10 normal individuals and 41 patients with various stages of gastric cancer. Soluble CD44S and its isoforms, V5 and V6, were present in the serum of normal individuals (288.53 +/- 18.33, 25.49 +/- 1.70, and 148.32 +/- 3.15 ng/ml, respectively). The concentrations of soluble CD44 V5 and V6 were elevated in patients with advanced gastric carcinoma (69.39 +/- 6.06 and 216.62 +/- 32.98 ng/ml, respectively). Serum CD44 V5 concentrations correlated with the extent of tumor invasion (T), the status of lymph node involvement (N), and distant metastasis (M) (TNM staging) (p < 0.05), whereas CD44S did not. These results suggest that detection of abnormal regulation of CD44 splicing could be helpful in gastric cancer diagnosis and disease evaluation.