Neuropeptides concentrations in the skin of a murine (NC/Nga mice) model of atopic dermatitis

BACKGROUND: It has been reported that the expression of neuropeptides (NPs), and the density and structure of peripheral nerves in atopic dermatitis (AD) are different from those in normal skin. OBJECTIVE: We investigated the role of NPs, in the development of AD with quantitative study of substance P (SP) and calcitonin gene-related peptide (CGRP) in the skin of AD-model mice. METHODS: We measured the NPs in the skin of mice (NC/Nga as AD-model mice, BALB/c and C57BL/6 as control) by enzyme-linked immunosorbentassay (ELISA). Peripheral nerve fibers and SP in the skin were stained by immunohistochemical staining, using anti-PGP9.5 antibody and anti-SP antibody. RESULTS: Under conventional condition, SP concentration in AD-like skin lesions of NC/Nga mice was higher than that in non-affected skin of the same mice. Under specific pathogen-free condition, SP concentration in the skin of NC/Nga mice was higher than that in the skin of BALB/c and C57BL/6 mice. In contrast, CGRP concentration in the skin lesions was lower than that in non-affected skin of NC/Nga mice. SP was detected not only in the nerve fibers in the dermis but also in mast cells in the inflammatory areas. CONCLUSIONS: The skin of NC/Nga mice contains more SP congenitally, and environmental factors may aggravate this abnormal condition. We hypothesize that increase of SP accompanied with a decrease of CGRP in the skin may play important roles in the pathogenesis and development of AD.     

Variations in early gross motor milestones and in the age of walking in Japanese children

Background: Gross motor development is usually assessed in terms of age of achievement of motor milestones. Although there is generally an impression of faster development if the milestones are achieved at younger ages, no longitudinal studies have been done on the associations between the milestones, especially in Japan. As a part of the Japan Children's Study, the purpose of the present study was to determine whether the achievement of gross motor milestones in infancy is related with the age of walking. Methods: This was a prospective cohort study of 290 healthy and term infants born in a district of Osaka City, Japan. Three milestones (rolling over, sitting, and crawling) were observed in the laboratory for infants aged at 4 and 9 months by a pediatrician and a developmental psychologist, and the age of walking was confirmed in questionnaires filled in by the parents at 18 and 27 months. Results: Children who could roll over at 4 months, and sit and crawl at 9 months, walked earlier than children who could not roll over, sit and crawl, respectively. With regard to crawling, children who were creeping had a 1 month delay in walking, and those who could not move forward had a 2 month delay compared to typical crawlers. On multiple regression analysis these three milestones were positively associated with walking: rolling over (β= 0.567), sitting (β= 1.973) and crawling (β= 1.473). Conclusion: The age and the patterns of sitting, crawling and rolling over were all related to the age of independent walking among Japanese infants. Consideration of milestone definition and variations is essential in medical check‐up.     

Preliminary study on evaluation of the pancreatic tail observable limit of transabdominal ultrasonography using a position sensor and CT-fusion image

Background and aim

Transabdominal ultrasonography (US) is commonly used for the initial screening of bilio-pancreatic diseases in Asian countries due to its widespread availability, the non-invasiveness and the cost-effectiveness. However, it is considered that US has limits to observe the area, namely the blind area. The observation of the pancreatic tail is particularly difficult. The goal of this study was to examine the pancreatic tail region that cannot be visualized on transverse scanning of the upper abdomen using US with spatial positional information and factors related to visualization, and observation of the tail from the splenic hilum.

Methods

Thirty-nine patients with pancreatic/biliary tract disease underwent CT and US with GPS-like technology and fusion imaging for measurement of the real pancreatic length and the predicted/real unobservable (PU and RU) length of the pancreatic tail. RU from US on transverse scanning and the real pancreatic length were used to determine the unobservable area (UA: RU/the real pancreatic length). Relationships of RU with physical and hematological variables that might influence visualization of the pancreatic tail were investigated.

Results

The real pancreatic length was 160.9 ± 16.4 mm, RU was 41.0 ± 17.8 mm, and UA was 25.3 ± 10.4%. RU was correlated with BMI (R = 0.446, P = 0.004) and waist circumferences (R = 0.354, P = 0.027), and strongly correlated with PU (R = 0.788, P < 0.001). The pancreatic tail was visible from the splenic hilum in 22 (56%) subjects and was completely identified in 13 (33%) subjects.

Conclusions

Combined GPS-like technology with fusion imaging was useful for the objective estimation of the pancreatic blind area.     

Translation,cultural adaptation,and validation of the American Skindex-29 quality of life index

BACKGROUND

Measuring the quality of life measure of patients with dermatologic diseases is an important concern. The instruments to evaluate it are commonly originally written in English and need to be translated and validated to be used in different cultures.

OBJECTIVE

The purpose of this paper is to translate and validate the Skindex-29 questionnaire to Brazilian Portuguese to be used in our country as a quality of life assessment instrument in dermatologic patients.

METHODS

The first step was the translation from English to Brazilian Portuguese and the back-translation by two native speakers. The translated version was then used for the second step, when three questionnaires were applied to 75 patients (43 of whom were classified as lightly affected and 32 as heavily affected by their dermatologic conditions): an identification questionnaire, the translated version of Skindex-29, and the Brazilian Portuguese version of Dermatologic Life Quality Index (DLQI). Additionally, the generic questionnaire Short Form 36 (SF-36) was applied to 41 of these patients. The last step to evaluate reproducibility was repeating the Skindex-29 questionnaire by the same researcher one week later in 44 patients.

RESULTS

Reliability was observed in global Skindex-29 scale (α=0.934), and its domains emotions (α=0.926), symptoms (α=0,702), and psycosocial functioning (α=0.860). The reproducibility showed high intraclass correlations. High intra class correlations was observed, thus validating reliability.

CONCLUSIONS

The Skindex-29 quality of life questionnaire was properly translated and validated to Brazilian Portuguese.     

An expression of within-plate uncertainty in sandwich ELISA

This paper puts forward a method to describe an equation of the within-plate uncertainty (relative standard deviation (R.S.D.) of measurements) as a function of analyte concentration in sandwich enzyme-linked immunosorbent assay (ELISA). A kit for thyroid stimulating hormone is taken as an example. The pipetting procedures of analyte solution and chromogen-substrate solution and absorbance inherent to the wells of a microplate are identified as major error sources and their variability is included as parameters in the uncertainty equation. These parameters can be determined by the experiments with distilled water. The theoretical R.S.D. is shown to be in good agreement with the results of the repeated experiments using the real samples. Since the theory gives a continuous plot of R.S.D. against concentration, the uncertainty structure of the ELISA kit can be recognized over a wide concentration range and the detection limit and quantitation range can easily be determined on the plot.     

Interactions between migraine and tension-type headache and alcohol drinking, alcohol flushing, and hangover in Japanese

The aim of the study was to investigate associations between headache types and alcohol drinking, alcohol flushing, and hangover. Alcohol consumption is inhibited by the presence of inactive aldehyde dehydrogenase-2 (ALDH2) whose carriers are susceptible to alcohol flushing and hangovers. We conducted a cross-sectional study of the 2,577 subjects (men/women: 1,018/1,559) who reported having ever experienced headaches unrelated to common colds and alcohol hangovers among 5,408 (2,778/2,630) Tokyo health checkup examinees. We used a questionnaire inquiring about current and past facial flushing after drinking a glass of beer which identifies the presence of inactive ALDH2 with a sensitivity and specificity of approximately 90%. Based on ICHD-II criteria migraine was diagnosed in 419 (75/344) subjects, and tension-type headache (TTH) in 613 (249/364). We classified the headaches of the remaining 1,545 (694/851) of headaches sufferers into the category “other headaches (OH)”. The migraineurs drank alcohol less frequently than the subjects with TTH among current/past alcohol flushers and than the subjects with OH regardless of flushing category. No such difference in drinking frequency was observed between TTH and OH. Current/past flushers drank alcohol less frequently than never flushers, and the likelihood that male migraineurs would avoid alcohol drinking than men with TTH or OH was stronger among current/past flushers than among never flushers. Flushers and women were more susceptible to hangover than never flushers and men, respectively, regardless of headache type. Among never flushers, women with migraine were more susceptible to hangover than women with OH. The difference in alcohol sensitivity may partly explain less alcohol consumption by migraineurs.     

New strain typing method with Sporothrix schenckii using mitochondrial DNA and polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) technique

The complete sequences of mitochondrial DNA (mtDNA) from two strains of different genotypes, American Type Culture Collection 10268 of mtDNA type 1 and KMU2025 of mtDNA type 4, were determined. These are circular molecules, 27 125 and 26 095 bp in length, respectively. The greatest difference between the two strains was found in the region encompassed by atp9 and cox2 genes, which was amplified with polymerase chain reaction (PCR) and used for preliminary restriction fragment length polymorphism (RFLP) analysis. Eight isolates of five mtDNA types were used and RFLP patterns obtained with the restriction enzyme AseI showed that this method seems to have greater discrimination power than the other PCR-RFLP typing method using internal transcribed spacer regions of nuclear DNA.     

Psychological stress as a risk factor for postoperative keloid recurrence

ObjectiveTo investigate psychological stress on the prognosis of the postoperative recurrence of keloids.MethodsPatients with keloids (n = 25), candidates for surgical resection and postoperative radiotherapy, had their psychological stress evaluated on the day before the surgical procedure. The parameters evaluated were pain and itching (Visual Numerical Scale), quality of life (Questionnaire QualiFibro/Cirurgia Plástica-UNIFESP), perceived stress (Perceived Stress Scale), depression and anxiety (Hospital Depression and Anxiety Scale), salivary cortisol and minimum and maximum galvanic skin responses (GSR) at rest and under stress (i.e., while the questionnaires were being filled out). Patients were evaluated during the 3rd, 6th, 9th and 12th months of postoperative care. During each return visit, two experts classified the lesions as non-recurrent and recurrent.ResultsThe recurrence group presented the greatest values in GSR during a stressful situation. The chance of recurrence increased by 34% at each increase of 1000 arbitrary units in maximum GSR during stress.ConclusionPsychological stress influenced the recurrence of keloids.     

FUS/TLS-immunoreactive neuronal and glial cell inclusions increase with disease duration in familial amyotrophic lateral sclerosis with an R521C FUS/TLS mutation

Basophilic inclusions (BIs) are pathological features of a subset of frontotemporal lobar degeneration disorders, including sporadic amyotrophic lateral sclerosis (ALS) and familial ALS (FALS). Mutations in the fused in sarcoma/translocated in liposarcoma (FUS/TLS) gene have recently been identified as a cause of FALS. The FUS/TLS-immunoreactive inclusions are consistently found in cases of frontotemporal lobar degeneration with BIs; however, the association between ALS cases with BIs and FUS/TLS accumulation is not well understood. We used immunohistochemistry to analyze 3 autopsy cases of FALS with the FUS/TLS mutation and with BIs using anti-FUS/TLS antibodies. The disease durations were 1, 3, and 9 years. As the disease duration becomes longer, there were broader distributions of neuronal and glial FUS/TLS-immunoreactive inclusions. As early as 1 year after the onset, BIs, neuronal cytoplasmic inclusions and glial cytoplasmic inclusions were found in the substantia nigra in addition to the anterior horn of the spinal cord. Glial cytoplasmic inclusions are found earlier and in a wider distribution than neuronal cytoplasmic inclusions. The distribution of FUS/TLS-immunoreactive inclusions in FUS/TLS-mutated FALS with BIs was broader than that of BIs alone, suggesting that the pathogenetic mechanism may have originated from the FUS/TLS proteinopathy.     

Selection of Heterogeneous Vancomycin-Intermediate Staphylococcus aureus by Imipenem

Vancomycin (VAN)-intermediate Staphylococcus aureus (VISA) and heterogeneous VISA (hVISA) isolates are considered to have emerged from VAN-susceptible S. aureus (VSSA) by spontaneous mutation during VAN exposure. We previously reported that laboratory mutant H14, obtained from VSSA strain ΔIP by exposure to imipenem (IPM), showed overexpression of the vraSR two-component system and a typical hVISA phenotype. In the present study, to elucidate the mechanism of VSSA conversion to hVISA, we further characterized strain H14 by determining its whole-genome sequence, morphology, cell wall synthetic activity, and gene expression. Genome sequencing revealed that H14 harbored a mutated vraS (designated vraS_H14) that caused an amino acid substitution (S₃₂₉→L). This mutation is different from the VraS mutation (N₅→I) identified in representative clinical hVISA strain Mu3. However, H14 exhibited a phenotype similar to that of Mu3, including heterogeneous resistance to VAN, enhanced cell wall synthetic activity, and vraSR overexpression. Replacement of the vraS gene of ΔIP with the mutated vraS_H14 gene confirmed that the S₃₂₉→L substitution was responsible for both the upregulation of vraSR and conversion to the hVISA phenotype. This conversion was also achieved by using the vraS gene of Mu3, which carries a mutation (N₅→I), but not with the native vraS gene of strain N315. Finally, we carried out a study to analyze the appearance of hVISA from VSSA by exposure of ΔIP to selective concentrations of VAN and beta-lactam antibiotics. A total of 8 and 5 hVISA isolates were detected among 50 isolates selected with VAN and IPM, respectively. Among the 13 hVISA mutants, mutation in vraSR was detected only in mutant strain H14, suggesting that additional mutational mechanisms can be responsible for evolution to the hVISA phenotype. We conclude that exposure not only to VAN but also to beta-lactams may select for reduced glycopeptide susceptibility in S. aureus.Methicillin (meticillin)-resistant Staphylococcus aureus (MRSA) is a major cause of serious nosocomial infections, and the emergence of virulent MRSA strains in the community is of particular concern (6). Vancomycin (VAN) still serves as the main therapeutic agent for infections caused by multiresistant MRSA strains (17). However, MRSA strains with various degrees of reduced susceptibility to glycopeptides, vancomycin-intermediate S. aureus (VISA) and heterogeneous VISA (hVISA) strains, have emerged among multidrug-resistant MRSA clinical strains (9, 16).Recently, we identified several genes that are overexpressed in VISA strain Mu50 and hVISA Mu3 compared to their levels of expression in their isogenic VAN-susceptible S. aureus (VSSA) strain, strain Mu50Ω (13); and among these, we found the overexpression of the vraSR two-component system (TCS), an upregulator of the S. aureus cell wall biosynthesis pathway (12, 13). We also demonstrated that the vraS gene is overexpressed in ΔIP-H14 (H14), a laboratory-derived hVISA strain obtained by selecting VSSA strain N315ΔIP (ΔIP) with 8 mg/liter of imipenem (IPM) (12) and showed that a single amino acid substitution in VraS was present in H14, Mu3, and Mu50 (10a).In the study described here, we further characterized hVISA strain H14, investigated the role of the vraS mutation on the phenotype of H14, and evaluated the rates of selection of hVISA from VSSA ΔIP following exposure to VAN and beta-lactam antibiotics.