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Background and Purpose: Elevation of glutamate, an excitatory amino acid, during inflammation and injury plays a crucial role in the reception and transmission of sensory information via ionotropic and metabotropic receptors. This study aimed to investigate the mechanisms underlying the biphasic effects of metabotropic glutamate mGlu5 receptor activation on responses to noxious heat.Experimental Approach: We assessed the effects of intraplantar quisqualate, a non-selective glutamate receptor agonist, on heat and mechanical pain behaviours in mice. In addition, the effects of quisqualate on the intracellular calcium response and on membrane currents mediated by TRPV1 channels, were examined in cultured dorsal root ganglion neurons from mice.Key Results: Activation of mGlu5 receptors in hind paw transiently increased, then decreased, the response to noxious heat. In sensory neurons, activation of mGlu5 receptors potentiated TRPV1-mediated intracellular calcium elevation, while terminating activation of mGlu5 receptors depressed it. TRPV1-induced currents were potentiated by activation of mGlu5 receptors under voltage clamp conditions and these disappeared after washout. However, voltage-gated calcium currents were inhibited by the mGlu5 receptor agonist, even after washout.Conclusions and Implications: These results suggest that, in sensory neurons, mGlu5 receptors biphasically modulate TRPV1-mediated intracellular calcium response via transient potentiation of TRPV1 channel-induced currents and persistent inhibition of voltage-gated calcium currents, contributing to heat hyper- and hypoalgesia. [ABSTRACT FROM AUTHOR]

BACKGROUND AND PURPOSE: Elevation of glutamate, an excitatory amino acid, during inflammation and injury plays a crucial role in the reception and transmission of sensory information via ionotropic and metabotropic receptors. This study aimed to investigate the mechanisms underlying the biphasic effects of metabotropic glutamate mGlu5 receptor activation on responses to noxious heat. EXPERIMENTAL APPROACH: We assessed the effects of intraplantar quisqualate, a non-selective glutamate receptor agonist, on heat and mechanical pain behaviours in mice. In addition, the effects of quisqualate on the intracellular calcium response and on membrane currents mediated by TRPV1 channels, were examined in cultured dorsal root ganglion neurons from mice. KEY RESULTS: Activation of mGlu5 receptors in hind paw transiently increased, then decreased, the response to noxious heat. In sensory neurons, activation of mGlu5 receptors potentiated TRPV1-mediated intracellular calcium elevation, while terminating activation of mGlu5 receptors depressed it. TRPV1-induced currents were potentiated by activation of mGlu5 receptors under voltage clamp conditions and these disappeared after washout. However, voltage-gated calcium currents were inhibited by the mGlu5 receptor agonist, even after washout. CONCLUSIONS AND IMPLICATIONS: These results suggest that, in sensory neurons, mGlu5 receptors biphasically modulate TRPV1-mediated intracellular calcium response via transient potentiation of TRPV1 channel-induced currents and persistent inhibition of voltage-gated calcium currents, contributing to heat hyper- and hypoalgesia. [ABSTRACT FROM AUTHOR]

Aims To describe the burden of diabetes mellitus and impaired fasting glucose in middle-aged residents (35-64 years) in an urban area of Sri Lanka. Methods A cross-sectional survey was conducted in the Ragama Medical Officer of Health area, from which 2986 participants (1349 men and 1637 women) were randomly selected from the electoral registry between January and December 2007. The participants underwent a physical examination and had their height, weight, waist and hip circumferences and blood pressure measured by trained personnel. Fasting blood samples were taken for measurement of glucose, HbA1c and lipids. The prevalence of diabetes (fasting plasma glucose > 7 mmol/l) and impaired fasting glycaemia (fasting plasma glucose 5.6-6.9 mmol/l) and major predictors of diabetes in Sri Lanka were estimated from the population-based data. Results Age-adjusted prevalence of diabetes mellitus in this urban population was 20.3% in men and 19.8% in women. Through the present screening, 263 patients with diabetes and 1262 with impaired fasting glucose levels were identified. The prevalence of newly detected diabetes was 35.7% of all patients with diabetes. Among patients with diabetes, only 23.8% were optimally controlled. In the regression models, high BMI, high waist circumference, high blood pressure and hypercholesterolaemia increased the fasting plasma glucose concentration, independent of age, sex and a family history of diabetes. Conclusions Our data demonstrate the heavy burden of diabetes in this urban population. Short- and long-term control strategies are required, not only for optimal therapy among those affected, but also for nationwide primary prevention of diabetes. [ABSTRACT FROM AUTHOR]

Background. The prevalence of metabolic syndrome (MetS) within individual cohorts varies with the definition used. The aim of this study was to compare the prevalence of MetS between IDF and revised NCEP ATP III criteria in an urban Sri Lankan population and to investigate the characteristics of discrepant cases. Methods. 2985 individuals, aged 35-65 years, were recruited to the study. Anthropometric and blood pressure measurements and laboratory investigations were carried out following standard protocols. Results. Age and sex-adjusted prevalences of MetS were 46.1% and 38.9% by revised NCEP and IDF definitions, respectively. IDF criteria failed to identify 21% of men and 7% of women identified by the revised NCEP criteria. The discrepant group had more adverse metabolic profiles despite having a lower waist circumference than those diagnosed by both criteria. Conclusion. MetS is common in this urban Sri Lankan cohort regardless of the definition used. The revised NCEP definition was more appropriate in identifying the metabolically abnormal but nonobese individuals, especially among the males predisposed to type 2 diabetes or cardiovascular disease. Further research is needed to determine the suitability of the currently accepted Asian-specific cut-offs for waist circumference in Sri Lankan adults. [ABSTRACT FROM AUTHOR]

Aim Colonoscopy may need to be rescheduled because of inadequate bowel preparation. We evaluated the effectiveness of colonoscopic enema as rescue for an inadequate 1-day bowel preparation before colonoscopy. Method Patients referred for afternoon colonoscopy were prospectively enrolled in the study during a 1-year period. Patients took bowel preparation (polyethylene glycol) solution on the morning of the endoscopy. If during colonoscopy the bowel preparation was poor, an enema of polyethylene glycol solution (500 ml) was instilled into the colon at the level of the hepatic flexure via the biopsy channel of the colonoscope which was then removed. The patient was allowed to recover from the propofol sedation and used the bathroom to evacuate the enema. The colonoscope was then introduced and the examination continued. Results Of 504 patients undergoing colonoscopy, 26 (4.9%) received an enema. The median age was 59 (29-79) years and 19 (73%) were female. A subsequent successful colonoscopy was achieved in 25/26 (96%). There were no complications. The mean time spent for the entire colonoscopy from the initial preparation to the end of the examination including the enema was 7.6 ± 1.1 h (5.4 h preparation, 0.2 h first colonoscopy + enema, 0.66 h waiting in the lavatory, 0.33 h second colonoscopy and 1 h for recovery). Conclusion Colonoscopic enema was highly successful as rescue for patients with inadequate bowel preparation and avoided postponement of the procedure. [ABSTRACT FROM AUTHOR]

Ancient glass vessel fragments belonging to the seventh to ninth centuries ad, from the Ko Kho Khao, Laem Pho and Khuan Lukpad sites in southern Thailand, were studied. The glass vessel fragment samples are a collection belonging to the Department of Archaeology, the 15th Regional Office of Fine Arts of Thailand. The chemical compositions of the glasses were analysed using a modified portable energy-dispersive X-ray fluorescence spectrometer (OURSTEX 100FA-II) by the introduction of a MOXTEK® AP3.3 polymer window (5 mm2∅) to the KETEK silicon drift detector for the measurement of light elements. The non-destructive analysis was performed at the National Museum, Phuket, in Thailand. It is shown that the glass chemical compositions belong to mineral and plant-ash based soda-lime-silicate glass. The origins of the glass artefacts are discussed in terms of raw materials and glass decoration, and compared with previously reported similar typological glasses from sites in the port city of Rāya and the Monastery of Wadi al-Tur in Egypt. [ABSTRACT FROM AUTHOR]

Aim Endoscopic decompression of malignant colorectal obstruction is often dealt with using expandable metallic stents. Endoscopic decompression of benign large bowel obstruction is more difficult. We report the technique and outcome of transanal endoscopic decompression for benign large bowel obstruction. Method From January 2001 to June 2010, endoscopic decompression using a transanal drainage tube placement was attempted in consecutive patients with benign large bowel obstruction. The clinical features, technical success, complications, treatment after the tube placement and clinical success were retrospectively evaluated. Results There were 13 patients (seven males, age 47-87, mean 69 years). The sites of obstruction were transverse colon [5 (38%)], sigmoid colon [3 (23%)], ileocecal valve [2 (15%)], splenic flexure [1 (8%)], descending colon [1 (8%)] and rectum [1 (8%)]. The most common cause of obstruction was anastomotic stricture [9 (69%)]. In 12 (92%) patients transanal decompression was technically successful with one perforation. An overtube, the reinsertion of colonoscope along the decompression tube, or the use of a small-diameter endoscope was required for the tube placement in seven (54%). In seven (54%) patients tube placement alone resulted in relief of bowel obstruction without operation. Conclusion Endoscopic decompression using a transanal drainage tube is effective for the management of benign large bowel obstruction. [ABSTRACT FROM AUTHOR]

N. Kato, K. Narutomi, M. Fukase and T. Motoyama Hollow spheroids in ascites of ovarian clear cell carcinoma: how are they formed and how do they behave? Objective: Although the multicellular aggregates (spheroids) in malignant ascites are usually solid throughout, they sometimes have acellular hollow spaces, especially in ascites of ovarian clear cell carcinoma. The purpose of this study is to analyse the origin and behaviour of hollow spheroids. Methods: Archival cytological and histological specimens of 32 ovarian carcinomas, including 12 clear cell carcinomas, were reviewed. HAC-2, a clear cell carcinoma cell line, was injected into the abdominal cavity of nude mice for direct comparison of ascitic cytology and tumour histology. Spheroids that were collected from nude mice ascites were cultured in vitro to observe their behaviour. Results: Five of six clear cell carcinomas with hollow spheroids showed spherule-like hyaluronan-rich stroma in their tumour tissue, whereas those without hollow spheroids did not. After heterotransplantation, both ascites and tumour imprints showed small or large hollow spheroids. Hyaluronan was detected in the former but not in the latter. The abdominal tumours showed compact spherule-like hyaluronan-rich stroma, enlarged oedematous stroma or intermediate stroma. In both size and hyaluronan status, small and large hollow spheroids were approximately comparable to spherule-like hyaluronan-rich stroma and oedematous stroma, respectively. During culture in vitro, hollow spheroids were maintained as hollow spheroids in suspension, and produced daughter hollow spheroids. Conclusions: The hollow space in the spheroids originates from spherule-like hyaluronan-rich stroma, where water trapping by hyaluronan causes enlargement of the space. The matrix within the hollow space serves as a scaffold that regulates cell polarity and matrix production. [ABSTRACT FROM AUTHOR]

Marui T, Funatogawa I, Koishi S, Yamamoto K, Matsumoto H, Hashimoto O, Jinde S, Nishida H, Sugiyama T, Kasai K, Watanabe K, Kano Y, Kato N. The NADH-ubiquinone oxidoreductase 1 alpha subcomplex 5 ( NDUFA5) gene variants are associated with autism. Autism appears to have a strong genetic component. The product of the NADH-ubiquinone oxidoreductase 1 alpha subcomplex 5 ( NDUFA5) gene is included in the mitochondrial electron transport chain. We performed a case-control study of 235 patients with autism and 214 controls and examined three single-nucleotide polymorphisms (SNPs) within this gene in a Japanese population. We then conducted a transmission disequilibrium test (TDT) analysis in 148 autistic trios. In the case-control study, two SNPs (rs12666974 and rs3779262) showed a significant association with autism ( P = 0.00064 and 0.00046 respectively). Furthermore, a haplotype containing these two SNPs showed a significant association ( P-global = 0.0013, individual haplotype : P = 0.010). In TDT analysis, the global and haplotype P-values also indicated significant associations. Minor allele and genotype frequencies were decreased in the autistic subjects. We found significant association between the NDFA5 gene and autism. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To investigate the molecular mechanisms underlying particle-induced osteolysis, we focused on osteopontin (OPN), a cytokine and cell-attachment protein that is associated with macrophage chemoattractant and osteoclast activation. METHODS: We compared OPN protein levels in human periprosthetic osteolysis tissues with those in osteoarthritis (OA) synovial tissues. To investigate the functions of OPN during particle-induced osteolysis in vivo, titanium particles were implanted onto the calvaria of OPN-deficient mice and their wild-type (WT) littermates. Mice were killed on day 10 and evaluated immunohistologically. The effects of OPN deficiency on the secretion of inflammatory cytokines were examined using cultured bone marrow-derived macrophages (BMMs). BMMs from OPN-deficient and WT mice were cultured with titanium particles for 12 hours, and the concentrations of inflammatory cytokines in the conditioned media were measured by enzyme-linked immunosorbent assay. RESULTS: Expression of OPN protein was enhanced in human periprosthetic osteolysis tissues as compared with OA synovial tissues. In the particle-induced model of osteolysis of the calvaria, bone resorption was significantly suppressed by OPN deficiency via inhibition of osteoclastogenesis, whereas an inflammatory reaction was observed regardless of the genotype. Results of immunostaining indicated that OPN protein was highly expressed in the membrane and bone surface at the area of bone resorption in WT mice. When BMMs were exposed to titanium particles, the concentration of proinflammatory cytokines, such as tumor necrosis factor alpha, interleukin-1alpha (IL-1alpha), IL-1beta, and IL-6, as well as chemotactic factors, such as monocyte chemoattractant protein 1 and macrophage inflammatory protein 1alpha, in the conditioned medium were significantly reduced by OPN deficiency. Whereas phagocytic activity of BMMs was not attenuated by OPN deficiency, phagocytosis-mediated NF-kappaB activation was impaired in OPN-deficient BMMs. These data indicated that OPN was implicated in the development of particle-induced osteolysis via the orchestration of pro-/antiinflammatory cytokines secreted from macrophages. CONCLUSION: OPN plays critical roles in wear debris-induced osteolysis, suggesting that OPN is a candidate therapeutic target for periprosthetic osteolysis. [ABSTRACT FROM AUTHOR]

It is controversial whether past hepatitis B virus infection constitutes an additional risk of hepatocellular carcinoma (HCC) among patients with hepatitis C virus (HCV). The incidence of HCC between 1994 and 2004 was analysed among 1262 patients who were only positive for HCV. The cumulative incidence of HCC was assessed by Kaplan–Meier analysis and the difference between two groups was assessed by the log-rank test. The effect of anti-HBc positivity on the risk of HCC was assessed with multivariate Cox proportional analysis. Anti-HBc was positive in 522 (41.4%) patients. The proportion of male patients (56.7 vs 46.8%, P < 0.001) and mean age (60.8 vs 56.9 years, P < 0.001) were significantly higher in the anti-HBc positive group. HCC developed in 339 patients (mean follow-up 7.0 years), with cumulative incidence rates at 3, 5 and 10 years of 12.7, 24.5 and 41.9% in the anti-HBc positive group and 10.6, 17.7 and 33.4% in the negative group, respectively ( P = 0.005). However, anti-HBc seropositivity did not reach statistical significance in multivariate analysis including age and gender (hazard ratio, 1.06; 95% CI, 0.85–1.31; P = 0.63). Anti-HBc positivity and HCC incidence were confounded by male gender and older age. [ABSTRACT FROM AUTHOR]

Macrophage migration inhibitory factor (MIF) may play an important role in the pathogenesis of acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT), as MIF plays an important role to regulate the production of tumor necrosis factor-α (TNF-α), one of the inflammatory cytokines which induces and exacerbates aGVHD. We examined the association between serum MIF levels and aGVHD vs. chronic GVHD (cGVHD) in allo-SCT patients in this study. We found a significant increase in the peak serum MIF (14.46 ng ± 1.47 ng/ml) at onset in patients that developed aGVHD ( n = 23, P = 0.009). We also found that mean serum MIF levels in patients who developed extensive type cGVHD within 6 months (12.58 ± 2.18 ng/ml, n = 13) were significantly higher than MIF levels before allo-HSCT (7.86 ± 1.17 ng/ml, n = 19, P = 0.04). Therefore, we speculated that serum MIF levels increase during the active phase of both aGVHD and cGVHD. [ABSTRACT FROM AUTHOR]

Background and Objective: The role of human leukocyte histocompatibility antigen (HLA) class II molecules on non-antigen-presenting cells has been a matter of controversy. We previously reported that HLA-II molecules on human gingival fibroblasts (GF) do not present antigens, but transduce signals into the cells, resulting in the expression of several cytokines, such as interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), regulated upon activation, normal T-cell expressed and secreted (RANTES) and IL-8. However, the exact role of these cytokines, as well as other cytokines which are potentially secreted from GF, in the pathogenesis of chronic periodontal inflammation is not fully understood. The aim of this study was to observe the effects of HLA-II-induced cytokines on the proliferation of human umbilical vein endothelial cells (HUVEC). Material and Methods: Antibody-based cytokine-microarray analyses were performed to detect potential cytokines associated with angiogenesis. Next, cytokine productivity was confirmed by quantitative methods. Then, cell proliferation assay was performed to see whether these cytokines promoted the proliferation of HUVEC. Results: Besides IL-6, MCP-1, RANTES and IL-8, growth-related gene product (GRO) was newly identified as an HLA-II-induced cytokine released from GF. This was confirmed by a quantitative method. Cell culture supernatant from HLA-II-stimulated GF cultures promoted the growth of HUVEC. Addition of anti-IL-8 neutralizing antibody, anti-CXC receptor (CXCR)1 antibody and anti-MCP-1 antibody inhibited the growth of HUVEC in a dose-dependent manner, while addition of anti-GROα antibody did not. Conclusion: The HLA-II-induced IL-8, via CXCR1, as well as MCP-1 from GF, promotes endothelial cell proliferation, which is possibly associated with enhanced angiogenesis in chronic periodontal lesions. [ABSTRACT FROM AUTHOR]

Aims:  Transcription factor hepatocyte nuclear factor (HNF)-1β is selectively expressed in clear cell carcinoma (CCC) of the ovary. One of the potential HNF-1β target genes is osteopontin ( OPN). Although elevation of OPN mRNA has been reported in CCC, it remains unclear whether CCC cells overexpress OPN protein. The aim was to investigate the expression of OPN protein and its correlation with HNF-1β status in CCC. Methods and results:  Three CCC and two serous adenocarcinoma (SA) cell lines were evaluated for expression of OPN by reverse transcriptase-polymerase chain reaction and immunocytochemistry. OPN expression, at both the mRNA and protein levels, was higher in the three CCCs than in the two SAs. HNF-1β expression was detected in the CCCs but not in the SAs. Subsequently, 60 surgical specimens (30 CCCs and 30 SAs) were examined immunohistochemically for expression of OPN and HNF-1β. All 30 CCCs showed immunopositivity for both OPN and HNF-1β. The 12 (40%) CCCs with a high OPN score all had a high HNF-1β score. In contrast, SAs rarely showed immunoreactivity for OPN or HNF-1β. Conclusions:  OPN expression is elevated in ovarian CCC and is closely associated with HNF-1β overexpression. HNF-1β is likely to participate in OPN up-regulation in CCC. [ABSTRACT FROM AUTHOR]

A 65-year-old Japanese male was diagnosed as multiple myeloma with Bence Jones kappa type, clinical stage IIIA. His disease status reached partial remission after chemotherapy. Thereafter, he received tandem transplantation, consisting of high-dose chemotherapy with autologous stem cell transplantation (ASCT), followed by unrelated cord blood transplantation (U-CBT). U-CBT with a reduced-intensity conditioning regimen (RI-CBT) was performed in August 2003. HLA mismatch between the patient and the CBT donor was present at two serological loci (B and DR). A total nucleated CBT cell dose of 2.45 × 107/kg body weight was infused on day 0. Graft- vs.-host disease (GVHD) prophylaxis consisted of cyclosporine A and short-term methotrexate. Neutrophil engraftment (>0.5 × 109/l) was obtained on day 46. He developed positive cytomegalovirus antigenemia, grade II acute GVHD involving skin and liver, varicella–zoster virus infection, septic shock, hemorrhagic cystitis caused by adenovirus and acute hepatitis B virus infection after U-CBT. We retrospectively analyzed T-cell receptor (TCR) repertoire diversity and found that TCR repertoire diversity decreased continuously after U-CBT. Therefore, low-TCR repertoire diversity in this patient appears to be associated with various infections caused by immunodeficiency. [ABSTRACT FROM AUTHOR]

Background An increased understanding of the ocular surface alterations at the cellular level in the conjunctiva and the cornea, may help explain the pathogenesis and the subsequent clinical appearance of atopic ocular allergies, which may be potentially blinding. Purpose To investigate MUC 1, 2 and 4 alterations, tear function and the ocular surface disorder in patients with atopic keratoconjunctivitis. Methods Twenty-eight eyes of 14 atopic keratoconjunctivitis patients as well as 22 eyes of 11 age-and sex-matched normal subjects were studied. The subjects underwent corneal sensitivity measurements, Schirmer's test, tear film break-up time (BUT), fluorescein and Rose Bengal staining of the ocular surface, conjunctival impression cytology and brush cytology. Impression cytology samples underwent periodic acid-Schiff and immunohistochemical staining with MUC 1, 2 and 4 antibodies. Brush cytology specimens underwent evaluation for inflammatory cell numbers and quantitative real-time-PCR for MUC 1, 2 and 4 mRNA expression. Patient eyes with fluorescein and Rose Bengal scores greater than four points were regarded to have significant epithelial disease in this study. Results The mean corneal sensitivity and BUT values were significantly lower in atopic patients with significant epithelial disease, compared with patients with insignificant epithelial disease and controls ( P<0.01). Brush cytology specimens from patients with significant epithelial disease revealed significantly higher numbers of inflammatory cells ( P<0.01). Specimens from patient eyes showed positive staining for MUC 1, 2 and 4. MUC 1, 2 and 4 mRNA expressions were significantly higher in eyes with significant epithelial disease compared with eyes with insignificant epithelial disease and eyes of control subjects. Conclusion Ocular surface inflammation, decline in corneal sensitivity, tear film instability, changes in conjunctival epithelial MUC 1, 2 and 4 mRNA expressions were thought to be important in the pathogenesis of atopic ocular surface disease. [ABSTRACT FROM AUTHOR]

Abstract: Objective: To identify adequate weight gain ranges during pregnancy in Japanese women. Method: Obstetric records from 2001 to 2002 for 46,659 term, singleton, vaginally delivered live births was used to estimate IUGR and macrosomia risk. Total maternal weight gain was grouped according to gestational age-specific percentile values of weight gain as follows: “very low” (under the 25th), “low” (25th to 49th), “moderate” (50th to 74th), “high” (75th to 89th), and “very high” (90th and over). Results: About 6% of infants were identified as having IUGR and 0.9% as macrosomia. IUGR risk was elevated with low weight gains. Macrosomia risk was related to high weight gains and previous spontaneous abortions. Conclusion: Achieving weight gains between the 50th and 75th percentiles for gestational age was considered adequate for optimal fetal growth in Japanese pregnant women. [Copyright &y& Elsevier]

Phosphohexomutases reversibly catalyse the transfer of the phosphate group of a glycosyl phosphate between the C6 and C1 positions, and uridine diphosphate (UDP)-hexose pyrophosphorylases catalyse the synthesis of UDP-hexose from uridine triphosphate (UTP) and hexose-1-phosphate. Both enzyme families are essential for nucleoside diphosphate hexose biosynthesis and are therefore critical for various physiological functions in the midgut of mosquitoes after a blood meal. We cloned and sequenced three phosphohexomutase and two UDP-hexose pyrophosphorylase cDNAs from Aedes aegypti. The products of the cDNAs were expressed and substrate specificities were examined. Herein we describe Ae. aegypti phosphoglucomutase 1, phosphoglucomutase 2, phosphoacetylglucosamine mutase, UDP-glucose pyrophosphorylase, and UDP- N-acetylglucosamine pyrophosphorylase. Transcripts of the genes expressing the enzymes are constitutively present in all life stages and blood-feeding does not seem to influence transcript abundance. [ABSTRACT FROM AUTHOR]

Kato N, Ohyama H, Nishimura F, Matsushita S, Takashiba S, Murayama Y. Role of helper T cells in the humoral immune responses against 53-kDa outer membrane protein fromPorphyromonas gingivalis.Oral Microbiol Immunol 2005: 20: 112–117.© Blackwell Munksgaard, 2005.Outer membrane protein with a 53-kDa molecular weight (Ag53) isolated fromPorphyromonas gingivalisevokes strong humoral immune responses in many periodontitis patients. To examine the effects of cytokines produced by Ag53-specific Th cells on the IgG production against Ag53, we established Ag53-specific Th-cell lines from patients with early onset periodontitis and from healthy volunteers. We then developed a mixed lymphocyte culture system between Ag53-specific Th cells and auto- or allo-derived T-cell-depleted leukocytes produced from the subjects whose HLA class IIhaplotypes were completely matched. Interferon-γ production was observed in all Th cell lines from patients and healthy subjects. As for Th2 type cytokines, interleukin (IL)-4, IL-5, IL-6 and IL-10 production varied greatly in Th cells regardless of the periodontal condition of the donor. Only Th cell lines with a high Th2/Th1 ratio induced Ag53-specific IgG production when cocultured with T-cell-depleted leukocytes. Thus, the difference in Th2/Th1 balance may regulate the Ag53-specific IgG production. [ABSTRACT FROM AUTHOR]

To clarify the birth length of twins according to gestational age.We studied a total of 51 910 live-birth–live-birth pairs of twins, 4561 triplet live births and 256 quadruplet live births, using data obtained from corresponding birth certificates. The birth length of twins was analyzed according to gestational age.Compared to singleton neonates, the median birth length of twins was approximately 0.5 cm smaller after the gestational age of 34 weeks, increasing to approximately 2.0 cm at 42 weeks of gestation. The median birth length according to gestational age was found to be the greatest in twins, lower in triplets and the lowest in quadruplets, in which the difference was<2.0 cm.The birth length of twins was smaller than that of singletons, but the difference was smaller than the difference in birthweight between twins and singletons. [ABSTRACT FROM AUTHOR]

Abstract Glucosamine:fructose-6-phosphate aminotransferase (GFAT) catalyses the formation of glucosamine 6-phosphate and is the first and rate-limiting enzyme of the hexosamine biosynthetic pathway. The final product of the hexosamine pathway, UDP-N -acetyl glucosamine, is an active precursor of numerous macromolecules containing amino sugars, including chitin in fungi and arthropods. Chitin is one of the essential components of insect cuticle and peritrophic matrix. The peritrophic matrix is produced in the midgut of mosquitoes in response to bloodfeeding, and may affect vector competence by serving as a physical barrier to pathogens. It is hypothesized that GFAT plays a regulatory role in biosynthesis of chitin and peritrophic matrix formation in insects. We cloned and sequenced the GFAT gene (AeGfat-1 ) and its 5′ regulatory region from Aedes aegypti . There is no intron in AeGfat-1 and there are two potential transcription start sites. AeGfat-1 cDNA is 3.4 kb in length and its putative translation product is 75.4 kDa. The amino acid sequence of GFAT is highly conserved in lower and higher eukaryotes, as well as in bacteria. AeGfat-1 message is constitutively expressed but is gradually up-regulated in the midgut after bloodfeeding. The putative regulatory region of the gene contains the ecdysone response element, E74, and Broad complex motifs, similar to what is found in the glutamine synthetase gene in Ae. aegypti. Results suggest that Ae. aegypti GFAT-1 may have a regulatory role in chitin biosynthesis and peritrophic matrix formation, and probably is under the regulation of ecdysteroids. [ABSTRACT FROM AUTHOR]

SummaryConcern about the healthy growth and development of an only child has been voiced since the 1970s, when the Chinese government launched the only child policy. In this study, the physical and personality traits of rural Chinese preschool only-children (onlies) whose age ranged from 3 to 6 years old were evaluated. The sample included 197 onlies and 367 children with siblings who came from seven kindergartens in rural areas in Fuzhou, Fujian province. The results showed no statistically significant differences in height, mass or BMI between the onlies and siblings. Regarding the personality traits, the significant difference was that the onlies exhibited more somatic complaints, however, the data didn’t indicate any other undesirable personality traits for the onlies. These results suggest that Chinese preschool children grow normally with or without siblings. [ABSTRACT FROM AUTHOR]

A case of a 63-year-old man with ameloblastoma with basal cell carcinoma (BCC)-like features clinically emerging as a nasal polyp is reported. The left nasal cavity was filled with a solid mass, which seemed to be a sinusitis-associated nasal polyp. The polyp was covered by parakeratotic squamous epithelium which was directly connected to the BCC-like tumor nest. The BCC-like features gradually changed to adamantinoid features. The polyp was connected with a huge mass filling the maxillary sinus and the molar area, which consisted of conventional ameloblastoma features. Although the tumor was finally diagnosed as an ameloblastoma of the maxilla, the biopsy specimen forced us to face the problem of differential diagnosis, ameloblastoma with BCC-like features or adamantinoid basal cell carcinoma (BCC). Immunohistochemical examination revealed that tumor cells of the ameloblastoma reacted with anticytokeratin antibody KL-1 but not with antiepithelial antibody Ber-EP4, and these reaction patterns were completely contrary to those of BCC. It is emphasized that immunohistochemical examination using anticytokeratin antibody KL-1 and antiepithelial antibody Ber-EP4 is a good tool for distinguishing ameloblastoma with BCC-like features from adamantinoid BCC. [ABSTRACT FROM AUTHOR]

Previously, we found that Kiebsiella O3 lipopolysaceharide (KO3 LPS) exhibits very strong adjuvant activity in augmenting the delayed-type hypersensitivity and antibody responses to protein antigens, and also strong ability to enlarge the regional lymph node in mice. By hydrolysis in 1% acetic acid at 100 for 1 hr. KO3 LPS was dissociated into 54-60% polysaccharide fraction and 25-27% lipid A fraction. The lipid A fraction thus prepared retained lethal toxicity for mice, whereas the polysaccharide fraction was essentially non-toxic. However, neither the lipid A fraction nor the polysaceharide fraction could reproduce the strong adjuvanticity of KO3 LPS, and a simple mixture of these two fractions also failed to do so. It is therefore concluded that, for expression of the strong adjuvanticity of KO3 LPS, both the lipid A and polysaccharide moieties are indispensable and must be combined in the form of LPS. By contrast, administration of relatively large amounts of the lipid A fraction alone could elicit enlargement of the regional lymph node to the same degree as that attained by KO3 LPS. The regional lymph node-enlarging activity of the lipid A fraction in lesser amounts was enhanced by addition of the polysaccharide fraction. It is therefore likely that there is no direct correlation between adjuvant activity of the LPS or its derivatives, and their regional lymph node-enlarging activity. [ABSTRACT FROM AUTHOR]

In mice primed with a mixture of bovine serum albumin (BSA) and adjuvant (capsular polysaccharide of Klebsiella pneumoniae (CPS-K)) cell-associated immunological memory was increased secondarily after a second injection of BSA alone, whereas a primary injection of BSA alone into normal unprimed mice did not result in detectable memory. The optimum antigen doses for expression of the primary and secondary memories of adoptively transferred cells from unboosted prime donors or boosted donors in vivo culture systems were very similar, although those observed in intact mice were very different, as reported previously. The size of the secondary memory of adoptively transferred cells from boosted donors was more than ten times greater than that of the primary memory of adoptively transferred cells from unboosted primed donors. The lag period for increase of the secondary memory was shorter than that for the primary memory. Both primary and secondary memories increased during a long period (up to 3 months) after the antigenic stimulus. From the results of this study it was concluded that cell-associated immunological memory could be amplified in a secondary fashion upon contact with a second stimulus.

Intramuscular injection of a non-immunogenic dose (1 mg) of the capsular polysaccharide of Klebsiella pneumoniae (CPS-K) into mice caused a marked and prolonged increase in the amount of the serum IgM and IgG. The increase of IgM after this injection was over 100 times greater than that of antigen-specific IgM antibodies produced by injection of an immunogenic dose of CPS-K (5 μg) or sheep red blood cells SRBC) (2 × 108). The increase in the amount of immunoglobulins was not related to induction of immunological paralysis to the major subcomponent of CPS-K (acidic CPS-K) by. a large dose of antigen, but was induced by non-antigenic stimulation by the minor subcomponent of CPS-K (neutral CPS-K), which was non-immunogenic at any dose. In mice injected with a non-immunogenic dose of CPS-K, antibody levels in tile serum to three kinds of non-cross-reacting antigens, SRBC, bovine serum albumin (BSA) and a possibly autochthonous IgG (IgM-IgG mixed type cryoglobulin) were also increased. The number of plaque-forming cells (PFC) for SRBC in the spleens of mice was also markedly increased after CPS-K injection. These antibodies to SRBC and BSA and PFC for SRBC were exclusively of IgM type and increased to as high levels as those found after specific antigenic stimulation. This marked increase in IgM antibody was neither followed by an increase in IgG antibody nor by immunological memory for a secondary IgG response. [ABSTRACT FROM AUTHOR]

Klebsiella 03 lipopolysaccharide (LPS) and Escherichia coli 09 LPS were previously shown to have potent adjuvant activities in augmenting antibody response and delayed-type hypersensitivity to protein antigens. The R form LPS extracted from the O-specific polysaccharide-less mutants derived from Klebsiella 03 and E. coli 09 strains were characterized by chemical and electrophoretic analyses. The adjuvant activities of the R-LPS in augmenting antibody response and DTH to protein antigens were much weaker than those of the parental LPS. The strength of the adjuvant activities of the R-LPS was similar to that of the activities of Salmonella minnesota LPS and Ra-LPS. By contrast, polyclonal B-cell activation effects of the R-LPS were stronger than those of the parental LPS. The common feature of the parental LPS is that their O-specific polysaccharides are mannose homopolymers (mannans). From these results, it is suggested that mannose homopolymers as the O side-chains of LPS contribute to the action of LPS in enhancing strongly the T-cell dependent immune responses to protein antigens. [ABSTRACT FROM AUTHOR]

The role of carrier in both elicitation of anti-hapten antibody responds and generation of hapten-specific memory i n chickens were studied using dinitrophenyl (DNP) conjugate of T-independent antigen carrier (DNP-Ficoll), that of isologous serum protein DNP-chicken serum albumin (CSA), and heavily conjugated DNP-bovine gammaglobulin (BGG), All of these DNP-conjugates could generate DNP specific memory as markedly as moderately conjugated DNP-BGG. Both DNP-Ficoll and DNP-CSA hardly elicited the primary and secondary anti-DNP antibody responses. Heavily conjugated DNP-BGG did not elicit a significant primary anti-DNP response, whereas it elicited as high secondary anti-DNP response as moderately conjugated DNP-BGG. It is suggested therefore that, when chickens are stimulated with hapten-carrier conjugates, hapten-specific memory is generated independently of T-cell function, but the elicitation of both the primary and secondary anti-hapten antibody responses is somehow relevant to T-cell function, although the T-cell dependence is less marked in the secondary than in the primary response. [ABSTRACT FROM AUTHOR]

Using the capsular polysaccharide of Kiebsiella pneumoniae (CPS-K) as a polyclonal B-cell activator (PBA) and sheep red blood cells (SRBC) as a T-dependent antigen, the correlation of the actions of PBA and T-dependent antigen on B cells in induction and amplification of immunological memory was studied. B-memory cell function, as judged by anti-SRBC responsiveness in vitro of spleen cells to CPS-K, was amplified by the secondary injection of SRBC into SRBC-primed mice, whereas it was decreased markedly by injection of CPS-K. When CPS-K was injected simultaneously with, or 1 or 2 days before the secondary injection of SRBC, B-memory cell function was also decreased markedly. On the other hand, CPS-K did not inhibit induction of B-memory cell function when injected simultaneously with the primary injection of SRBC. However, CPS-K inhibited induction of B-memory cell function when injected 3 days before the primary injection of SRBC. The inhibition by CPS-K of amplification of B-memory cell function in response to SRBC when CPS-K was injected simultaneously with the secondary injection of SRBC occurred markedly in mice primed with SRBC 8 days or longer before the secondary injection, whereas it was not detectable in mice primed 3 days before. It is concluded that the CPS-K-mediated signal and the SRBC-mediated signal act competitively on the same subpopulations of B cells in induction and amplification of memory, and that the susceptibility of B cells to the CPS-K-mediated negative signal changes correspondingly with their maturation stage. [ABSTRACT FROM AUTHOR]

We have demonstrated that the number of rosette-forming cells (RFC) in the spleens of mice primed with sheep red blood cells (SRBC) was markedly decreased by administration of a polyclonal B-cell activator (PBA) such as the capsular polysaccharide of Klebsiella pneumoniae (CPS-K). Most of the RFC estimated were shown to be of B-cell type with Ig-receptors specific for SRBC. The precursor activity for the generation of anti-SRBC antibody-forming cells (AFC) (plaque-forming cells (PFC)) was closely associated with these RFC. Moreover, the precursor activity for the generation of AFC of RFC in the spleens of mice primed with SRBC and then treated with CPS-K (SRBC-primed and CPS-K-treated mice), as estimated by anti-SRBC PFC responsiveness in vitro to CPS-K, was much less than that of the same number of RFC in the spleens of SRBC-primed mice not treated with CPS-K especially at an early stage after injection of CPS-K. This low anti-SRBC PFC responsiveness of individual RFC in the spleens of SRBC-primed and CPS-K-treated mice resulted neither from an increase in some suppressing activity in the spleens of these mice nor from a relative increase in the number of RFC of non-B-cell type or non-SRBC-specific RFC. The percentage of the number of rosette-forming PFC in the total number of RFC seemed to be slightly increased in SRBC-primed and CPS-K-treated mice. However, this cannot totally explain the mechanism of the low responsiveness of RFC-fraction of spleen cells from SRBC-primed and CPS-K-treated mice. It has been concluded from these results that the signal mediated by PBA such as CPS-K acts on B cells bearing Ig-receptors specific for antigen (RFC) and changes a large number of them to the cells lacking Ig-receptors (non-RFC) and at least in part to the cells bearing Ig-receptors but showing low responsiveness to generate AFC following further stimulus (modified RFC). [ABSTRACT FROM AUTHOR]

Using the capsular polysaccharide of Klebsiella pneumoniae (CPS-K) as a polyclonal B-cell activator (PBA) and sheep red blood cells (SRBC) as a T cell-dependent antigen, we compared the ability of PBA and antigen to differentiate (generate antibody-foxing cells, AFC) and proliferate (generate immunological memory) virgin B cells and B memory cells. In vitro CPS-K induced the differentiation of IgM virgin B cells, IgM B memory cells and IgG B memory cells to AFC, as well as or better than SRBC. The differentiation of B memory cells to AFC by CPS-K did not require the participation of macrophages or T cells, whereas the action of SRBC depended strictly upon the helper actions of these cells. The responsiveness to CPS-K and SRBC of normal and antigen-primed spleen cells as judged by anti-SRBC PFC responses in vitro was markedly decreased after stimulation of virgin B cells and B memory cells in vivo by CPS-K injection into normal or primed mice but greatly increased after the injection of SRBC. The decrease in the responsiveness to CPS-K of spleen cells from mice treated with CPS-K appeared principally due to exhaustion of the functions of B cells and B memory cells. From the present data it has been concluded that the signals required for the differentiation and proliferation of B cells or B memory cells are different from each other, the signal for differentiation being provided by either antigen (SRBC) or PBA (CPS-K), while the signal for proliferation is delivered only by antigen. [ABSTRACT FROM AUTHOR]

We have cloned a 13 kb Escherichia coli DNA fragment which complemented the rfe mutation to recover the biosynthesis of E. coli O9 polysaccharide. Using Tn5 insertion inactivation, the rfe gene was localized at the 1.5kb HindIII-EcoRI region flanking the rho gene. We constructed an rfe-deficient E. coli K-12 mutant by site-directed inactivation using a DNA fragment of the cloned 1.5 kb rfe gene. This also confirmed the presence of the rfe gene in the 1.5kb region. By simultaneous introduction of both the rfe plasmid and the plasmid of our previously cloned E. coli 09 rfb into this rfe mutant, we succeeded in achieving in vivo reconstitution of O9 polysaccharide biosynthesis. From sequence analysis of the rfe gene, a putative promoter followed by an open reading frame (ORF) was identified downstream of the rho gene. This ORF coincided with the position of the rfe gene determined by Tn5 analysis and site-directed mutagenesis. Furthermore, we identified the rff genes in the 10.5kb DNA flanking the rfe gene. We recognized at least two functional domains on this cloned rff region. Region I complemented a newly found K-12 rff mutant, A238, to synthesize the enterobacterial common antigen (ECA). Deletion of region II resulted in the synthesis of ECAs with shorter sugar chains. When the 10.5kb rff genes of the plasmid were inactivated by either deletion or Tn5 insertion, the plasmid lost its ability to give rise to transformants of the rfe mutants. [ABSTRACT FROM AUTHOR]

Comparative studies were made of the increase in the numbers of plaque-forming cells (PFC), rosette-forming cells (RFC) and haemolytic foci for erythrocyte antigens in the spleens of mice given a non-specific stimulus (the capsular polysaccharide of Klebsiella pneumoniae (CPS-K)) and an antigenic stimulus (sheep red blood cells (SRBC)). The number of direct PFC for SRBC was increased by injection of CPS-K to as high a level as that obtained by injection of SRBC. In contrast, by injection of CPS-K the numbers of indirect PFC, RFC (probably the antibody-forming cell precursors) and haemolytic foci were not increased significantly, whereas all of them were increased markedly by injection of SRBC. The maximal number of PFC in mice injected with CPS-K was approximated to the number of background RFC of the same mice. Injection of CPS-K generated 25-130 times more direct PFC for each of three kinds of erythrocyte antigens, SRBC, rabbit red blood cells and chick red blood cells, than background PFC, whereas the total number of spleen cells was not increased significantly or increased very slightly. Repeated injections of CPS-K were not significantly more effective for increase in the number of direct PFC than a single injection of CPS-K. Injection of CPS-K could generate many direct PFC in mice which had been thymectomized, irradiated and reconstituted with foetal liver cells. In mice injected with CPS-K, increase in (or maintenance of) the numbers of direct PFC and RFC were inhibited by injection of a mitogen inhibitor, vinblastine sulphate, but their sensitivities to the drug were less than those found in mice immunized with SRBC. It has been concluded from these results that in mice injected with CPS-K a large number of antibody-forming cell precursors are differentiated to direct PFC through one division or a few divisions of the individual cells, and that the inability of CPS-K to induce sufficient cell divisions of the individual precursor cells is the cause of the lack of increase in the number of indirect PFC and in immunological memory for secondary PFC responses in mice injected with CPS-K. [ABSTRACT FROM AUTHOR]

Injection of bovine serum albumin (BSA) with the capsular polysaccharide of Klebsiella pneumoniae (CPS-K) as adjuvant strongly primes mice for booster injections of BSA alone. The best doses of antigen for booster injections varied according to the frequency of antigen injections and the time interval between antigen injections. Increase in memory following the injection of BSA mixed with CPS-K adjuvant was first detected 10 days later. Memory continued to increase as the time interval after priming was increased to 90 days. Memory for the tertiary response increased after the optimum secondary antigenic stimulus. Increase in memory after the secondary antigenic stimulus was detected as early as 7 days after injection. Even secondary injection of the high dose of 50 mg of BSA did not exhaust immunological memory. Memory was also increased after the tertiary antigenic stimulus. Extremely high antibody responses (up to 492 mg protein per ml of antibody) could be obtained by eliciting tertiary and quaternary antibody responses optimally at the time memory was developed to the maximum level. [ABSTRACT FROM AUTHOR]