Immunohistochemical Localization of the Aquaporins AQP1, AQP3, AQP4, and AQP5 in the Mouse Respiratory System

Aquaporins are membrane water channel proteins that function mainly in water transfer across cellular membranes. In our present study, we investigated the immunohistochemical distribution of aquaporin 1 (AQP1), AQP3, AQP4, and AQP5 in the mouse respiratory system by immunofluorescence, immunoperoxidase, and immunoelectron microscopy. AQP3, AQP4, and AQP5 are expressed in epithelial cells, whereas AQP1 is expressed in subepithelial connective tissues and capillaries. In the airway surface epithelia from the nasal cavity to the intrapulmonary bronchioles, AQP5 was found to be mainly localized to the luminal side and both AQP3 and AQP4 to the abluminal side. In the alveolar epithelium, AQP5 is localized to the apical membranes of both type I and type II alveolar cells. Compared with the previous studies on the rat respiratory system, in which AQP5 is restricted to the alveolar type I cells and absent from the airway surface epithelia, we found that AQP5 in the mouse is much more widely distributed throughout the surface epithelia. These results suggest that AQP5 has a critical role in water-handling, such as the maintenance of airway surface liquid and clearance of alveolar fluid in the mouse respiratory system

Differences in strategies for coping with menopausal symptoms in full-time workers and part-time workers in Japan

Working hours and working conditions are different for full-time workers and part-time workers. In this study, it was hypothesized that the strategies for coping with menopausal symptoms, such as hot flashes, night sweats and insomnia, and the proportion of women receiving clinical care differ between full-time and part-time workers. Four hundred and eighty-seven Japanese female workers aged 45 to 60 years responded to a health questionnaire about their understanding of menopause and their strategies for coping with menopausal symptoms. Based on the surveyed responses, the proportions of women with menopausal symptoms were 43.0% (80/186) in full-time workers and 48.2% (145/301) in part-time workers. There was no significant difference in the female worker response rate, nor were there significant differences in the proportions of full-time workers and part-time workers who had an understanding of menopause causes and treatments and who were able to cope with menopausal-related symptoms. However, there were significantly more full-time workers than part-time workers who received routine clinical care and who visited a hospital. On the other hand, significantly more part-time workers than full-time workers exercised and took dietary supplements as a means of coping with their symptoms. There are differences between female full-time and part-time workers regarding the strategies employed for coping with menopausal symptoms. The proportion of women receiving clinical care for their symptoms is greater among full-time employees than part-time employees. A physical examination can be a valuable opportunity for working women with menopausal symptoms to receive advice from a doctor or a nurse practitioner

Recent Advances in Fluorescent Labeling Techniques for Fluorescence Microscopy

Tremendous progress in recent computer-controlled systems for fluorescence and laser-confocal microscopy has provided us with powerful tools to visualize and analyze molecular events in the cells. Various fluorescent staining and labeling techniques have also been developed to be used with these powerful instruments. Fluorescent proteins such as green fluorescent protein (GFP) allow us to directly label particular proteins of interest in living cells. This technique has been extended over a large area of cell biology, and a variety of fluorescent protein-derived techniques have been developed to visualize the functions and conditions of the molecules within living cells. In this review, we summarize the techniques for fluorescent staining and labeling for recent fluorescence microscopy

Localization of Myosin and Actin in the Pelage and Whisker Hair Follicles of Rat

The combined effects of myosin II and actin enable muscle and nonmuscle cells to generate forces required for muscle contraction, cell division, cell migration, cellular morphological changes, the maintenance of cellular tension and polarity, and so on. However, except for the case of muscle contraction, the details are poorly understood. We focus on nonmuscle myosin and actin in the formation and maintenance of hair and skin, which include highly active processes in mammalian life with respect to the cellular proliferation, differentiation, and movement. The localization of nonmuscle myosin II and actin in neonatal rat dorsal skin, mystacial pad, hair follicles, and vibrissal follicles was studied by immunohistochemical technique to provide the basis for the elucidation of the roles of these proteins. Specificities of the antibodies were verified by using samples from the relevant tissues and subjecting them to immunoblotting test prior to morphological analyses. The myosin and actin were abundant and colocalized in the spinous and granular layers but scarce in the basal layer of the dorsal and mystacial epidermis. In hair and vibrissal follicles, nonmuscle myosin and actin were colocalized in the outer root sheath and some hair matrix cells adjoining dermal papillae. In contrast, most areas of the inner root sheath and hair matrix appeared to comprise very small amounts of myosin and actin. Hair shaft may comprise significant myosin during the course of its keratinization. These results suggest that the actin-myosin system plays a part in cell movement, differentiation, protection and other key functions of skin and hair cells

日本の看護師における更年期症状と仕事関連ストレス要因との関連

Objective: The main objective was to ascertain the typical menopausal symptoms and job-related stress factors in Japanese nurses during the menopausal transition, and the associations of menopausal symptoms with job-related stress. A supplementary objective was to determine whether there were any differences in menopausal symptoms and job-related stress factors among nurses in managerial positions. Methods: One thousand seven hundred female registered nurses aged 45–60 years who were working in hospitals in Japan were asked to complete a self-administered survey that included Greene’s Climacteric Scale and the Brief Job Stress Questionnaire. Results: The proportions of nurses who reported feelings of tiredness, irritability and difficulty in concentration were higher than the proportions with other menopausal symptoms. The proportions of nurses reporting feeling unhappy or depressed and having crying spells were higher among nurses in managerial positions than among other nurses. Stresses related to ‘quantitative overload’ on the Brief Job Stress Questionnaire among nurses in managerial positions were significantly greater than among nurses not in managerial positions, while stresses related to ‘physical overload’, ‘job control’, ‘skill discretion’, ‘workplace environment’ and ‘job satisfaction’ among nurses not in managerial positions were significantly greater than they were among nurses in managerial positions. Psychological symptoms were significantly correlated with poor job-related interpersonal relationships. Conclusions: Health care practitioners should be aware that menopausal symptoms are associated with job-related stress during the menopausal transition. Information on the differences in these associations between nurses in managerial positions and other nurses is important as it will allow their health care to be managed on a more individual basis

Molecular cloning and characterization of the human p19INK4d gene promoter

Abstractp19INK4d, a member of the INK4 family of cyclin-dependent kinase (CDK) inhibitors, negatively regulates the cyclin D–CDK4/6 complexes, which promote G1/S transition by phosphorylating the retinoblastoma tumor-suppressor gene product. To investigate the mechanism of transcriptional regulation of the p19INK4d gene, we characterized the 5′-flanking region of the human p19INK4d gene. The cap-site hunting method revealed that the transcription starts at −16 nucleotide (nt) upstream of the initiation codon. The 5′-flanking region of the human p19INK4d gene was ligated to a luciferase reporter gene and possessed functional promoter activity. Luciferase assay with a series of truncated 5′-flanking regions indicated that the region from −81 to −2 nt could drive the transcription of the p19INK4d gene. Several Sp1 and activating protein 2 binding sites are located within the region from −81 to −2 nt. Mutation of the second Sp1 binding site from −33 to −25 nt decreased the promoter activity. Collectively, it was demonstrated that the human p19INK4d gene is under the control of TATA-less promoter and the Sp1 binding site is involved in the transcription

Localization of Reversion-Induced LIM Protein (RIL) in the Rat Central Nervous System

Reversion-induced LIM protein (RIL) is a member of the ALP (actinin-associated LIM protein) subfamily of the PDZ/LIM protein family. RIL serves as an adaptor protein and seems to regulate cytoskeletons. Immunoblotting suggested that RIL is concentrated in the astrocytes in the central nervous system. We then examined the expression and localization of RIL in the rat central nervous system and compared it with that of water channel aquaporin 4 (AQP4). RIL was concentrated in the cells of ependyma lining the ventricles in the brain and the central canal in the spinal cord. In most parts of the central nervous system, RIL was expressed in the astrocytes that expressed AQP4. Double-labeling studies showed that RIL was concentrated in the cytoplasm of astrocytes where glial fibrillary acidic protein was enriched as well as in the AQP4-enriched regions such as the endfeet or glia limitans. RIL was also present in some neurons such as Purkinje cells in the cerebellum and some neurons in the brain stem. Differential expression of RIL suggests that it may be involved in the regulation of the central nervous system

MEGADOCK 3.0: a high-performance protein-protein interaction prediction software using hybrid parallel computing for petascale supercomputing environments

BACKGROUND: Protein-protein interaction (PPI) plays a core role in cellular functions. Massively parallel supercomputing systems have been actively developed over the past few years, which enable large-scale biological problems to be solved, such as PPI network prediction based on tertiary structures. RESULTS: We have developed a high throughput and ultra-fast PPI prediction system based on rigid docking, “MEGADOCK”, by employing a hybrid parallelization (MPI/OpenMP) technique assuming usages on massively parallel supercomputing systems. MEGADOCK displays significantly faster processing speed in the rigid-body docking process that leads to full utilization of protein tertiary structural data for large-scale and network-level problems in systems biology. Moreover, the system was scalable as shown by measurements carried out on two supercomputing environments. We then conducted prediction of biological PPI networks using the post-docking analysis. CONCLUSIONS: We present a new protein-protein docking engine aimed at exhaustive docking of mega-order numbers of protein pairs. The system was shown to be scalable by running on thousands of nodes. The software package is available at: http://www.bi.cs.titech.ac.jp/megadock/k/