Search

Salt intake reduction is a global concern. In particular, Japanese consume higher amounts of salt than those of other ethnicities. The sodium content is mentioned on the label of industrially prepared dishes with an intention of reducing salt intake. This study aimed to evaluate the difference between the actual sodium content and labeled salt value of industrially prepared Japanese single dishes. Samples labeled "estimated" were collected and classified as Japanese, Western, and Chinese cuisines. The sodium content ranged from 180 to 1011 mg/100 g. The sodium content was higher than their reported values in other countries. Specifically, Chinese dishes contained high amounts of sodium, although the chloride content was similar across cuisine styles. Further, the molar ratio (i.e., sodium/chloride) had no significant effect on the difference between the actual content and labeled value. The measured salt contents were 20% higher than the labeled values. The results of decision tree analysis indicated that if the labeled salt value of stir‐fried foods is determined by calculation, the actual sodium content is much higher than the labeled salt value. These findings are crucial for customers, dietitian, and researchers as they refer to the labeled salt value to determine the sodium content of industrially prepared foods. [ABSTRACT FROM AUTHOR]

Simple Summary: In the abdominal testes of most mammals, with the exception of elephants and a few animal species, spermatogenesis is characterised by an arrest of spermatogenesis due to heat stress. However, the mechanisms underlying normal spermatogenesis in the elephant abdominal testis remain unknown. We hypothesised that comparing the expression of heat stress tolerance molecules of elephant testes with that of other animals and investigating the mechanism of spermatogenesis in elephants might provide clues to overcome the causes of heat stress-induced spermatogenesis dysfunction. The results showed that the elephant's body cells, other than the testes, have a similar mechanism to those of other mammals but that the spermatogenic cells have a different mechanism, with different molecular immunoexpression under heat stress in the abdomen. These findings suggest that these molecules and their relationships may be important in future investigations of the mechanisms that overcome heat stress to maintain spermatogenesis in elephants. The abdominal testes of Asian elephants show normal spermatogenesis. Heat shock in cryptorchid testes elevates heat shock factor (HSF) expression, leading to germ cell apoptosis, while increased heat shock proteins (HSPs) levels provide protection. To investigate how heat shock affects elephant spermatogenic cells, focusing on heat shock-related molecules and the cell death mechanism, immunohistochemistry and TUNEL staining were employed to assess the immunoexpression of several heat shock-related molecules and the status of apoptosis in elephant fibroblasts (EF) induced by heat shock stimulus. Additionally, the immunoexpression of heat shock-related molecules and cell proliferation status in the elephant spermatogenic cells. Our finding indicated that heat shock-induced HSF1 immunoexpression in EF leads to apoptosis mediated by T-cell death-associated gene 51 (TDAG51) while also upregulating HSP70 to protect damaged cells. In elephant spermatogenic cells, immunostaining revealed a predominance of proliferating cell nuclear antigen (PCNA)-positive cells with minimal TDAG51- and TUNEL-positive cells, suggesting active proliferation and apoptosis suppression during normal spermatogenesis in the abdominal testis. Interestingly, spermatogonia co-immunoexpressed HSF1 and HSP90, potentially reducing apoptosis through protective mechanisms different from those observed in other mammals. Spermatogenic cells did not show immunolocalisation of HSP70, and hence, it may not contribute to protecting the spermatogonia from heat shock because the transcriptional activity of HSF1 is suppressed by HSP90A binding. This study provides insight into the specific heat shock response and defence mechanisms in elephant spermatogenic cells and may contribute to our understanding of species-specific adaptation to environmental stresses of the testis. [ABSTRACT FROM AUTHOR]

Multiple‐site optical recordings with NK2761, a voltage‐sensitive absorption dye, were applied to the embryonic chick olfactory system, and the functional development of olfactory nerve (N.I)‐related neural circuits was examined in the forebrain. The stimulation of the N. I elicited neural responses in N.I‐olfactory bulb (OB)‐forebrain preparations at the embryonic 8–12 day (E8‐E12) stages. At the E11 stage, we functionally identified two circuits projecting from the OB to the forebrain. The first circuit passed through the ventral side of the forebrain and spread in the dorso‐caudal direction, whereas the second circuit passed through the dorsal side to the first circuit. Pharmacological experiments showed that N‐methyl‐D‐aspartate (NMDA) receptor function was more significant for the transfer of sensory information in these circuits. The functional development of N.I‐related circuits was investigated, and the results obtained revealed that the ventral circuit was generated earlier than the dorsal circuit. Neural responses in the ventral circuit were detected from the E9 stage in normal physiological solution and the E8 stage in Mg2+‐free solution, which activated NMDA receptor function. At the E10 stage, neural responses in the dorsal circuit were clearly recognised in addition to ventral responses. We attempted to identify possible candidates for relay nuclei in the forebrain by comparing contour line maps of the optical signal amplitude with previously reported neuroanatomical data. The results suggest that N.I‐related neural circuits from the periphery to the subpallium functionally mature earlier than those to the pallium during ontogenesis. [ABSTRACT FROM AUTHOR]

Simple Summary: Diagnosis of neurodegenerative diseases requires examination of a variety of characteristics. A definitive diagnosis is obtained using a comprehensive evaluation of family history, neurological findings, brain imaging, genetic testing, and other medical information. Multiple-system atrophy (MSA) is a neurodegenerative disease associated with autonomic dysfunction, parkinsonism, and cerebellar ataxia, and early diagnosis is difficult because the disease changes over time. The aims of this study were to examine whether machine learning can improve diagnostic accuracy using MSA case data from a national survey, and to identify the features that are important for differentiation among MSA subtypes using machine learning. Multiple-system atrophy (MSA) is primarily an autonomic disorder with parkinsonism or cerebellar ataxia. Clinical diagnosis of MSA at an early stage is challenging because the symptoms change over the course of the disease. Recently, various artificial intelligence-based programs have been developed to improve the diagnostic accuracy of neurodegenerative diseases, but most are limited to the evaluation of diagnostic imaging. In this study, we examined the validity of diagnosis of MSA using a pointwise linear model (deep learning-based method). The goal of the study was to identify features associated with disease differentiation that were found to be important in deep learning. A total of 3377 registered MSA cases from FY2004 to FY2008 were used to train the model. The diagnostic probabilities of SND (striatonigral degeneration), SDS (Shy-Drager syndrome), and OPCA (olivopontocerebellar atrophy) were estimated to be 0.852 ± 0.107, 0.650 ± 0.235, and 0.858 ± 0.270, respectively. In the pointwise linear model used to identify and visualize features involved in individual subtypes, autonomic dysfunction was found to be a more prominent component of SDS compared to SND and OPCA. Similarly, respiratory failure was identified as a characteristic of SDS, dysphagia was identified as a characteristic of SND, and brain-stem atrophy was identified as a characteristic of OPCA. [ABSTRACT FROM AUTHOR]

Pregnancy and postpartum are periods in which women develop psychosocially. However, becoming a mother is stressful, and mood disorders related to anxiety and depression often develop. In recent years, research on interoception—sensations related to the body's internal physiological state—has attracted attention. Interoception has multifaceted characteristics. It involves directly perceiving information in the body while also inferring and evaluating it. In this study, we examined interoception, anxiety, and depression in Japanese pregnant women. Empirical examinations and questionnaire surveys were used to measure interoception in 32 pregnant women not at high risk of pregnancy. A Japanese adaption of the Multidimensional Assessment of Interoceptive Awareness was used to measure interoceptive sensibility, and a heartbeat counting task performance was used to measure interoceptive accuracy. Anxiety and depression were measured using the Japanese versions of the State-Trait Anxiety Inventory and the Edinburgh Postnatal Depression Scale, respectively. A correlation analysis was performed between interoception, anxiety and depression and between differences between sensibility and accuracy of interoception, anxiety and depression. We revealed that interoceptive sensibility and differences between sensibility and accuracy of interoception were associated with anxiety. Based on results of this pilot study, it is necessary to investigate using longitudinal studies whether interoception might be an effective predictor tool for early detection of anxiety during pregnancy and postpartum. [ABSTRACT FROM AUTHOR]

Some gastric epithelial neoplasms show predominant chief cell differentiation (oxyntic gland neoplasms), in which the entity of "gastric adenocarcinoma of fundic gland type" was firstly designated, whereas a possible more aggressive subgroup "gastric adenocarcinoma of fundic gland mucosa type" (GA-FGM) was subsequently proposed. However, the histopathologic progression mode of these neoplasms has not been sufficiently reported. In this article, we describe a case of GA-FGM in which we could observe its progression during 5 years. The tumor was removed by endoscopic submucosal dissection 5 years after the first biopsy, which had already shown a feature of oxyntic gland neoplasm. During the follow-up period, the endoscopy revealed little change in the tumor appearance. However, the histology of endoscopic submucosal dissection showed submucosal extension with its histological progression. Besides, other oxyntic gland neoplasms of the stomach were observed metachronously or synchronously, giving an implication about a common pathogenetic basis of these lesions. [ABSTRACT FROM AUTHOR]

Objectives: Dry socket and post-extraction pain are typical discomforts experienced by patients after tooth extraction. In this study, we inserted gauze coated with oxytetracycline-hydrocortisone ointment into the extraction socket immediately after lower third molar extraction and then evaluated the occurrence of dry socket and post-extraction pain compared with gauze non-insertion. Methods: This retrospective study was carried out on patients undergoing lower third molar extraction in the Department of Oral Surgery at Shizuoka Prefectural General Hospital in Shizuoka, Japan from November 2018 to October 2019. A comparison was carried out between a gauze-insertion group and a non-insertion group. The occurrence versus non-occurrence of dry socket was determined, and degree of pain was assessed based on a visual analogue scale (VAS) and on patients reporting the number of loxoprofen sodium oral analgesic tablets (60mg/tablet) that they had taken. Dry socket was defined as patient-reported spontaneous pain that did not subside 1 to 3 days postoperatively. Spontaneous post-extraction pain was recorded four times: on the operative day, on the first postoperative day (POD1), on POD3, and during suture removal (POD7). Results: The occurrence of dry socket was lower in the gauze-insertion group than in the non-insertion group (0.9%, 2/215 vs. 19.6%, 9/46, p<0.001). The results also showed that both VAS-defined pain level and the number of analgesic tablets taken were lower in the gauze-insertion group than in the non-insertion group on POD3 and POD7. Conclusions and clinical relevance: Inserting gauze coated with oxytetracycline-hydrocortisone ointment into the extraction socket immediately after third molar extraction reduces the occurrence of both dry socket and post-extraction pain. [ABSTRACT FROM AUTHOR]

In Mongolia, yak (Bos grunniens) are able to live in alpine areas and their products greatly influence the lives of the local people. Increased vigour in hybridized yak and cattle can offer benefits for livestock farmers. However, male hybrids show reproductive defects resulting from spermatogenesis arrest, affecting the conservation and maintenance of dominant traits in the next generation. The underlying mechanisms involved in hybrid cattle–yak infertility have recently been investigated; however, the genetic cause is still unclear. Androgens and androgen receptor (AR) signalling are required for spermatogenesis. We, therefore, evaluated the expression of AR, 3β‐hydroxysteroid dehydrogenase (3βHSD) and 5α‐reductase 2 (SRD5A2) in Leydig cells to investigate their function in cattle–yak spermatogenesis. Testicular tissues from yaks (1–3 years old) and hybrids (F1–F3, 2 years old) were collected and subjected to immunohistochemistry and image analyses to investigate the expression of each parameter in the Leydig cells. After maturation at 2 years, the expression levels of AR increased and the levels of 3βHSD decreased, but the SRD5A2 levels remained constant in yak. However, the cattle–yak hybrid F2 showed immature testicular development and significantly different expression levels of AR and 3βHSD compared with mature yak. These results suggest that the decreased expression of AR and increased expression of 3βHSD in the Leydig cells of cattle–yak hybrid testes may represent one of the causes of infertility. Our study might help in solving the problem of infertility in crossbreeding. [ABSTRACT FROM AUTHOR]

Facial motor neurons of the rat embryo are first generated in rhombomere 4 and then migrate in the caudo‐ventral direction. This migration forms a unique axonal trajectory called the genu, a loop of facial motor axons around the abducens nucleus. It is still unclear when and how this unique structure is functionally established during ontogenesis. Using voltage‐sensitive dye (VSD) recording and the DiI staining method, we identified neural responses evoked by facial nerve (N.VII) stimulation and examined developmental processes of the facial motor nucleus in E12‐E17 rat brainstems. We identified two types of fast spike‐like signals; a long‐duration signal, which corresponded to the action potential in the N.VII soma, and a short‐duration signal, which reflected the action potential in the N.VII axons. The long‐duration signal was detected as early as E13, suggesting that the N.VII motor neuron is already excitable at the beginning of cell migration. The response area of the long‐duration signal extended caudally at E13‐E14, and shifted in a ventral direction at E15. At E16‐E17, the long‐duration signal was concentrated in the caudo‐ventral area, which was comparable to the location of the facial motor nucleus in the adult rat brainstem. These results demonstrate that developmental processes of cell migration and nuclear organization can be visualized and identified functionally with the VSD recording. We discuss the results by comparing functiogenesis and morphogenesis of the N.VII pathway. Appling voltage‐sensitive dye recording and the DiI staining method to the embryonic rat brainstem, we examined functiogenesis/morphogenesis of the facial motor system. Stimulation of the facial nerve elicited a long‐duration signal, which reflected the activity in the soma, and a short‐duration signal, which was associated with the axonal pathway. By examining the spatiotemporal patterns of these signals, we elucidated the developmental dynamics and functional organization of the facial motor nucleus. [ABSTRACT FROM AUTHOR]

Objectives: Considering that there is a lack of evidence regarding the contribution of library and information services to evidence-based medicine in actual clinical practice in Japan, the purpose of the study is to explore the current status of use and value of library and information services in clinical settings to examine the usefulness of information in implementing evidence-based medicine (EBM) into practice. Methods: A Web-based survey was conducted at seven sites (hospitals with 300–1,200 beds) and interviews conducted at five sites to investigate information behavior among health professionals (physicians, residents, and nurses) in 2016, replicating the Value Study carried out in the United States in 2010 and 2011. Using a critical incident technique, respondents answered questions about their information topics, information resources used, search location, access points, and evaluation of the information. Results: Analysis from 598 valid responses (275 physicians, 55 residents, and 268 nurses) revealed the characteristics of information use and recognition of the value of information. Physicians and residents showed their information needs regarding clinical care using PubMed (80.4%, 65.5%), Ichushi-Web (61.8%, 63.6%), and UpToDate (40.4%, 65.5%). While physicians rely more on electronic journals (37.8%), residents use more hybrid resources including Japanese print books (38.2%) and online books (30.9% for Japanese, 32.7% for English) to confirm their knowledge. Nurses need more information close to patients and explore a wider variety of information resources such as Japanese print books (60.4%), Ichushi -Web (40.3%), Japanese online books (20.5%), and websites of academic organizations (19.0%). Although the overall recognition of the value of information was relatively modest, concrete changes in clinical practice were found in some areas. Environments with insufficient information and availability of electronic resources should be improved to increase the use of library and information services for implementing EBM. [ABSTRACT FROM AUTHOR]

One of the earliest activities expressed within the developing central nervous system is a widely propagating wave-like activity, which we referred to as the depolarization wave. Despite considerable consensus concerning the global features of the activity, its physiological role is yet to be clarified. The depolarization wave is expressed during a specific period of functional synaptogenesis, and this developmental profile has led to the hypothesis that the wave plays some roles in synaptic network organization. In the present study, we tested this hypothesis by inhibiting the depolarization wave in ovo and examining its effects on the development of functional synapses in vagus nerve-related brainstem nuclei of the chick embryo. Chronic inhibition of the depolarization wave had no significant effect on the developmental time course, amplitude, and spatial distribution of monosynaptic excitatory postsynaptic potentials in the first-order nuclei of the vagal sensory pathway (the nucleus of the tractus solitarius (NTS) and the contralateral non-NTS region), but reduced polysynaptic responses in the higher-order nucleus (the parabrachial nucleus). These results suggest that the depolarization wave plays an important role in the initial process of functional synaptic expression in the brainstem, especially in the higher-order nucleus of the cranial sensory pathway. [ABSTRACT FROM AUTHOR]

• We optically detected oscillatory activity in the embryonic chick olfactory bulb. • Olfactory nerve stimulation induced the oscillation following the EPSP. • The short-term perfusion of a Ca2+-free solution blocked the EPSP and oscillation. • The long-term perfusion of a Ca2+-free solution newly induced oscillatory activity. • The patterns of the oscillatory activity are different between with and without Ca2+. We applied 464/1020-site optical recording systems with a voltage-sensitive dye (NK2761) to the embryonic chick olfactory system and detected oscillatory activity in the olfactory bulb (OB) in the absence of synaptic transmission. In embryonic day 8–10 (E8-E10) chick olfactory nerve (N.I)-OB-forebrain preparations, the removal of Ca2+ from the external solution completely blocked the glutamatergic excitatory postsynaptic potential (EPSP) from the N.I to the OB as well as oscillations following the EPSP. However, a novel type of oscillatory activity was detected in the OB with the long-term perfusion of a Ca2+-free solution. The characteristics of oscillatory activity in the Ca2+-free solution differed from those in normal physiological solution. The present results suggest the existence of a neural communication system in the absence of synaptic transmission at the early stage of embryonic development. [ABSTRACT FROM AUTHOR]

The influences of soaking treatment and storage conditions on the softening of cooked beans, namely, soybeans and red kidney beans, were investigated. It was revealed that the softening of fresh soybeans and fresh red kidney beans was suppressed during subsequent boiling after soaking treatment at 50 and 60 °C. Furthermore, in treated aged soybeans and red kidney beans that were subjected to storage at 30 °C/75% relative humidity for 6 mo and soaking treatment at 50 to 60 °C, the hardness during cooking was further amplified. This suggested that the mechanism of softening suppression differs depending on the influences of soaking and storage. Analysis of the pectin fraction in alcohol insoluble solid showed insolubilization of metal ions upon storage at high temperature and high humidity in both soybeans and red kidney beans, which suggests interaction between Ca ions and hemicellulose or cellulose as cell wall polysaccharides. The results of differential scanning calorimetry (DSC) showed that aged soybeans exhibited a shift in the thermal transition temperature of glycinin-based protein to a higher temperature compared with fresh soybeans. From the results of DSC and scanning electron microscopy for aged red kidney beans, damaged starch is not conspicuous in the raw state after storage but is abundant upon soaking treatment. As for the influence of soaking at 60 °C, it can be suggested that its influence on cell wall crosslinking was large in soybeans and red kidney beans in both a fresh state and an aged state. [ABSTRACT FROM AUTHOR]

Purpose: Biphasic cuirass ventilation (BCV) is a form of non-invasive extrathoracic positive and negative pressure mechanical ventilation. The present study was conducted to quantify our positive experience using BCV to dramatically improve gas exchange and cardiac function in patients with acute exacerbation of chronic respiratory failure and secondary pulmonary hypertension (PH). Methods: BCV was applied for 2 weeks in 17 patients with PH caused by lung disease. Ventilation sessions were limited to 1 h per day to prevent exhaustion. To assess respiratory and circulatory effects, percutaneous arterial oxygen saturation (SpO) was measured before and after each daily BCV session, and right heart catheter test [mean pulmonary artery pressure (mPAP), right atrium pressure (RAP), pulmonary artery occlusion pressure (PAOP) and cardiac index (CI)] and serum N-terminal pro-brain natriuretic peptide (NT-proBNP) were measured before and after a series of BCV sessions. Results: SpO transiently improved after each BCV session. After a series of BCV, mPAP decreased from 27.2 to 22.4 mmHg ( p = 0.0007). PAOP, CI and serum NT-proBNP levels decreased compared with baseline. No patients were treated with epoprostenol, iloprost, bosentan or sildenafil for PH. Conclusion: BCV may improve circulatory function in patients with PH caused by lung disease. [ABSTRACT FROM AUTHOR]

The central issue in developmental neuroscience is when and how neural synaptic networks are established and become functional within the central nervous system ( CNS). Investigations of the neural network organization have been hampered because conventional electrophysiological means have some technical limitations. In this study, the multiple-site optical recording technique with a voltage-sensitive dye was employed to survey the developmental organization of the vagal system in the mouse embryo. Stimulation of the vagus nerve in E11-E14 mouse embryos elicited optical responses in areas corresponding to the vagal sensory and motor nuclei. Postsynaptic responses in the first-order sensory nucleus, the nucleus of the tractus solitarius ( NTS), were identified from E11, suggesting that sensory information becomes transferred to the brain at this stage. In addition to the NTS, optical responses were identified in the rostral and contralateral brainstem regions, which corresponded to second/higher order nuclei of the vagus nerve including the parabrachial nucleus ( PBN). Postsynaptic responses in the second/higher-order nuclei were detected from E12, suggesting that polysynaptic networks were functional at this stage. We discuss the results of our optical mapping, comparing them with previous findings obtained in the chick and rat embryos, and suggest some fundamental principles in the functional organization of synaptic networks in the embryonic brain. [ABSTRACT FROM AUTHOR]

BlastKOALA and GhostKOALA are automatic annotation servers for genome and metagenome sequences, which perform KO (KEGG Orthology) assignments to characterize individual gene functions and reconstruct KEGG pathways, BRITE hierarchies and KEGG modules to infer high-level functions of the organism or the ecosystem. Both servers are made freely available at the KEGG Web site ( http://www.kegg.jp/blastkoala/ ). In BlastKOALA, the KO assignment is performed by a modified version of the internally used KOALA algorithm after the BLAST search against a non-redundant dataset of pangenome sequences at the species, genus or family level, which is generated from the KEGG GENES database by retaining the KO content of each taxonomic category. In GhostKOALA, which utilizes more rapid GHOSTX for database search and is suitable for metagenome annotation, the pangenome dataset is supplemented with Cd-hit clusters including those for viral genes. The result files may be downloaded and manipulated for further KEGG Mapper analysis, such as comparative pathway analysis using multiple BlastKOALA results. [ABSTRACT FROM AUTHOR]

Background: Long-term cancer prognosis after initial surgical procedures is an unlikely endpoint for clinical trials. Medical claim databases may aid in addressing this issue regardless of limited information on disease and patient background. However, the long-term prognosis (especially regarding long-term care needs) following surgical procedures remains unclear. This study aimed to assess whether long-term outcomes, such as the exacerbation of long-term care needs and mortality, differ with surgical methods. Methods: Using a longitudinal study with linkage between medical claim and long-term care database, patients with primary colorectal cancer who underwent initial colonoscopies were identified through anonymized data in Japan (Shizuoka Kokuho Database, 2012–2018). Odds ratios (ORs) for long-term outcomes (long-term care needs and all-cause mortality during a 6.5-year follow-up period) were analyzed using logistic regression to compare laparoscopy and endoscopic surgery to laparotomy. Results: Overall, 3,744 primary colorectal cancer cases (822 laparotomies, 705 laparoscopies, and 2,217 endoscopic surgeries) were included. Compared to the laparotomy group, the crude OR for exacerbation of long-term care needs in the laparoscopic surgery group was 0.376 (95% confidence interval, 0.227, 0.624), while the OR for all-cause mortality was 0.22 (0.329, 0.532). Conclusion: This is the first study to analyze long-term prognosis after surgery for patients with colorectal cancer to combine medical and long-term needs data. As the national health insurance claim database rarely includes information on cancer stage and comorbidities, better prognosis on endoscopic surgery may need careful interpretation. Therefore, laparoscopy has superior outcomes in terms of long-term care needs and mortality compared to those of laparotomy. [ABSTRACT FROM AUTHOR]

Highlights: [•] We examined the initial expression of synaptic function in the embryonic chick CNS. [•] We applied voltage-sensitive dye imaging to the N.V-brainstem preparation. [•] We optically identified N.V-related sensory and motor nuclei in the brainstem. [•] Synaptic function in the sensory nuclei emerged earlier than previously considered. [•] Development of synaptic function differs among nerves and between sensory nuclei. [ABSTRACT FROM AUTHOR]

Potassium (K+)-uptake transport proteins present in prokaryote and eukaryote cells are categorized into two classes; Trk/Ktr/HKT, K+ channel, and Kdp belong to the same superfamily, whereas the remaining K+-uptake family, Kup/HAK/KT has no homology to the others, and neither its membrane topology nor crucial residues for K+ uptake have been identified. We examined the topology of Kup from Escherichia coli. Results from the reporter fusion and cysteine labeling assays support a model with 12 membrane-spanning domains. A model for proton-coupled K+ uptake mediated by Kup has been proposed. However, this study did not show any stimulation of Kup activity at low pH and any evidence of involvement of the three His in Kup-mediated K+ uptake. Moreover, replacement of all four cysteines of Kup with serine did not abolish K+ transport activity. To gain insight on crucial residues of Kup-mediated K+ uptake activity, we focused on acidic residues in the predicted external and transmembrane regions, and identified four residues in the membrane regions required for K+ uptake activity. This is different from no membrane-localized acidic residues essential for Trk/Ktr/HKTs, K+ channels and Kdp. Taken together, these results demonstrate that Kup belongs to a distinct type of K+ transport system. [ABSTRACT FROM AUTHOR]

This study investigated the effects of skim milk on the quality and fertility of boar spermatozoa under long-term chilled preservation. Semen samples were stored in Modena solution supplemented with 0 (control) to 50 mg/mL skim milk at 5° C for 4 weeks; spermatozoa stored with 7.5 and 15 mg/mL of skim milk (7.5- SM and 15- SM groups, respectively) exhibited significantly higher motility indices than those of the control group up to 3 weeks ( P < 0.05), and the 7.5- SM group showed improved motility indices even after 4 weeks ( P < 0.05). In vitro fertilization using spermatozoa in the 7.5- SM and 15- SM groups stored at 5° C for 2 weeks showed significantly higher fertilization rates of spermatozoa and the development rates to blastocyst than the control group ( P < 0.05), and the 7.5- SM group showed similar rates of fertilization and blastocyst formation in the fresh non-stored spermatozoa group. After artificial insemination using spermatozoa stored for 2 weeks in the 7.5- SM group, healthy piglets were obtained. Boar spermatozoa can be stored at 5° C in a Modena solution containing skim milk. Supplementation of 7.5 mg/mL skim milk improves boar spermatozoa motility and fertility even after liquid preservation at 5° C for 2 weeks. [ABSTRACT FROM AUTHOR]

We investigated the characteristics of dietary supplements and their use by 1,076 Japanese pregnant women, the majority of whom were in mid- to late pregnancy. The subjects completed a self-reported survey on their sociodemographic characteristics, supplement use, and attitudes towards diet. The overall prevalence of supplement use did not change before and after pregnancy (75%); however, daily use increased by approximately two-fold with pregnancy (20.2% versus 37.2%). After the onset of pregnancy, supplements containing folic acid were taken for fetal health. Daily users were more likely to be older, have a greater awareness of the risk of neural tube defects (NTD), view supplement use as acceptable, have less diet anxiety, and have more advisers regarding diet. Respondents used supplements containing folio acid alone or with other ingredients. Folic acid intake is recommended to reduce the risk of NTD. However, supplement use began after pregnancy recognition, suggesting a lack of knowledge on the appropriate timing of folic acid use. Information about supplements was obtained mostly from newspapers, magazines, flyers, and stores. These results indicate that more accurate information regarding the optimal timing of folic acid intake and the safety of dietary supplements must be disseminated. [ABSTRACT FROM AUTHOR]

Abstract: The multiple-site optical recording technique with a voltage-sensitive dye, NK2761, was used to survey the functional organization of neuronal networks related to the vagus nerve (N.X) in the E16-stage rat brainstem. When we stimulated N.X, in addition to the responses in the vagal sensory nucleus (nucleus of the tractus solitarius (NTS)) on the stimulated side, other response areas were bilaterally detected. Characteristics of the optical signals in these areas suggested that they correspond to neural activity in the second/higher-ordered nucleus of the vagal pathway. The first area was located at the level of the pons. Based upon morphological information, we suggest that this area corresponds to the parabrachial nucleus (PBN), which receives inputs from the NTS. The second area was located between the NTS and the PBN. We suggest that this area is the A5 noradrenergic group. These results suggest that the N.X-related neural networks are established similarly to the adult pattern from an early developmental stage. [Copyright &y& Elsevier]

Spontaneous embryonic movements, called embryonic motility, are produced by correlated spontaneous activity in the cranial and spinal nerves, which is driven by brainstem and spinal networks. Using optical imaging with a voltage-sensitive dye, we revealed previously in the chick and rat embryos that this correlated activity is a widely propagating wave of neural depolarization, which we termed the depolarization wave. One important consideration is whether a depolarization wave with similar characteristics occurs in other species, especially in different mammals. Here, we provide evidence for the existence of the depolarization wave in the mouse embryo by summarizing spatiotemporal characteristics and pharmacological natures of the widely propagating wave activity. The findings show that a synchronized wave with common characteristics is expressed in different species, suggesting its fundamental roles in neural development. [ABSTRACT FROM AUTHOR]

Although constipation is very common during pregnancy, the exact mechanism is unknown. We hypothesized that the involvement of estrogen receptor (ER) in the regulation of electrolyte transporter in the colon leads to constipation. In this study, the intestines of normal female ICR mouse and pregnant mice were examined for the expression of ERα and ERβ by immunohistochemistry and in situ hybridization. ERβ, but not ERα, was expressed in surface epithelial cells of the proximal, but not distal, colon on pregnancy days 10, 15, and 18, but not day 5, and the number of ERβ-positive cells increased significantly during pregnancy. Expression of NHE3, the gene that harbors estrogen response element, examined by immunohistochemistry and western blotting, was localized in the surface epithelial cells of the proximal colon and increased in parallel with ERβ expression. In ovariectomized mice, NHE3 expression was only marginal and was up-regulated after treatment with 17β-estradiol (E), but not E + ICI 182,780 (estrogen receptor antagonist). Moreover, knock-down of ERβ expression by electroporetically transfected siRNA resulted in a significant reduction of NHE3 expression. These results indicate that ERβ regulates the expression of NHE3 in the proximal colon of pregnant mice through estrogen action, suggesting the involvement of increased sodium absorption by up-regulated NHE3 in constipation during pregnancy. [ABSTRACT FROM AUTHOR]

During the early development of the nervous system, synchronized activity is observed in a variety of structures, and is considered to play a fundamental role in neural development. One of the most striking examples of such activity is the depolarization wave reported in chick and rat embryos. In the accompanying paper (Momose-Sato et al., 2012), we have demonstrated that a depolarization wave is also present in the mouse embryo by showing large-scale optical waves, which spread remarkably over the central nervous system, including the spinal cord, hindbrain, cerebellum, midbrain, and forebrain. In the present study, we examined the pharmacological nature of the mouse depolarization wave and its developmental changes. We show here that two types of switching in pharmacological characteristics occur during development. One is that the depolarization wave is strongly dependent on nicotinic acetylcholine receptors during the early developmental stage [embryonic day (E)11-12], but is dominated by glutamate at the later stage (E13 onwards). The second is that γ-aminobutyric acid (GABA), which acts as an excitatory mediator of the depolarization wave during the early phase, becomes an inhibitory modulator by E14. These changes seemed to occur earlier in the hindbrain than in the spinal cord. Furthermore, we show that the second switch causes the loss of synchronization over the network, resulting in the disappearance of the depolarization wave and segregation of the activity into discrete regions of the medulla and spinal cord. We suggest that pharmacological switching is a possible mechanism underlying replacement of the primordial correlated network by a mature neuronal circuit. [ABSTRACT FROM AUTHOR]

Spontaneous embryonic movements, called embryonic motility, are produced by correlated spontaneous activity in the cranial and spinal nerves, which is driven by brainstem and spinal networks. Using optical imaging with a voltage-sensitive dye, we have revealed previously that this correlated activity is a widely propagating wave of neural depolarization, which we termed the depolarization wave. We have observed in the chick and rat embryos that the activity spread over an extensive region of the CNS, including the spinal cord, hindbrain, cerebellum, midbrain and forebrain. One important consideration is whether a depolarization wave with similar characteristics occurs in other species, especially in different mammals. Here, we provide evidence for the existence of the depolarization wave in the mouse embryo by showing that the widely propagating wave appeared independently of the localized spontaneous activity detected previously with Ca2+ imaging. Furthermore, we mapped the origin of the depolarization wave and revealed that the wave generator moved from the rostral spinal cord to the caudal cord as development proceeded, and was later replaced with mature rhythmogenerators. The present study, together with an accompanying paper that describes pharmacological properties of the mouse depolarization wave, shows that a synchronized wave with common characteristics is expressed in different species, suggesting fundamental roles in neural development. [ABSTRACT FROM AUTHOR]

BACKGROUND The aim of this study was to evaluate the histochemical characteristics of the thickened inner acellular layer (IL) of the lamina propria specifically present in the human seminiferous tubules of testes showing impaired spermatogenesis. METHODS Eighteen biopsies for the investigation of infertility and 10 orchiectomies for testicular cancer and cryptorchidism were used. Lectin staining [peanut agglutinin (PNA), Maackia amurensis (MAA), Sambuccus nigra (SNA)], PNA lectin staining with sialidase digestion, immunohistochemistry and binding assay of progesterone were performed and analysed quantitatively. RESULTS The IL of the thickened lamina propria of the seminiferous tubules in the testes showed PNA lectin affinity and binding affinity for progesterone. Both affinities of MAA and SNA were in the IL of only fairly thickened lamina propria. Furthermore, a positive correlation was present between the thickness of the lamina propria and the accumulation of glycoconjugates showing PNA lectin affinity (r = 0.829, P < 0.001) or progesterone (r = 0.629, P < 0.001) in the IL. However, ILs show no immunoreactivities of progesterone receptor, androgen receptor or human serum albumin. Progesterone inhibited the binding affinity of PNA lectin to the IL (P < 0.001), but not the affinity to the spermatogenic cells. In addition, sialidase digestion increased the PNA affinity not in the IL but in the spermatogenic cells (P < 0.001). CONCLUSIONS These results indicate that the IL of the thickened lamina propria always consists of glycoconjugates with PNA lectin affinity and possible binding affinity to progesterone. In addition, the glycoconjugates in the IL may be predictors of abnormal spermatogenesis in the testes of infertile patients. [ABSTRACT FROM AUTHOR]

We compared ascorbic acid (AA) levels in the blood and TPA- and fMLP-stimulated superoxide (O2•−) production in neutrophils of pre-, early, and late hypertensive stroke-prone spontaneously hypertensive rats (SHRSP) with those of age-matched Wistar Kyoto rats (WKY), or two other normotensive strains of rats. Plasma and lymphocyte AA levels were about two-fold higher in SHRSP as early as 4 weeks old compared to WKY, and also higher than those of Wistar and Sprague-Dawley (SD) rats. Levels of AA were high in the liver and adrenal glands of SHRSP, indicating congenitally high AA levels. The production of O2•− in neutrophils was about two-fold higher in SHRSP than in WKY even at 4 weeks of age, and increased with age in both strains. Among SHRSP, AA levels in lymphocytes decreased at the late hypertensive stages with a decrease in hepatic l-gulono-γ-lactone oxidase (GLO) activities. These data suggest that bi-phasic AA levels in the blood of SHRSP comprise congenitally high levels and a decrease after persistent hypertension due to enhanced O2•− production and a decrease in de novo AA synthesis through GLO. [ABSTRACT FROM AUTHOR]

Abstract: To regulate the autonomic function, the vagus nerve transfers various sensory information from peripheral organs, and appropriate motor reflexes are produced in the neural circuit. The functional development of the vagal pathway during the early phase of embryonic development has long been unclear. Optical recording with voltage-sensitive dyes has provided a new approach to the analysis of the functional development of the embryonic central nervous system. In this review, we present recent progress in optical studies on the vagal pathway in the embryonic chick and rat brainstems. The topics include how neural excitability is initially expressed in the motor and sensory nuclei [e.g. the dorsal motor nucleus of the vagus nerve (DMNV) and the nucleus of the tractus solitarius (NTS)] and how synapse networks are formed in the primary and higher-ordered sensory nuclei [e.g. the parabrachial nucleus (PBN)]. We also refer to the functional development of the glossopharyngeal nuclei and compare the developmental steps with those of the vagal nuclei. [Copyright &y& Elsevier]

Abstract: Collapsin response mediator protein2 (CRMP2) is a brain-specific protein involved in neuronal polarity and axonal guidance, and phosphorylation of CRMP2 regulates the function and the activity. CRMP2 has shown to be implicated in several neurodegenerative diseases (Alzheimer''s disease, epilepsy and ischemia) and this study was designed to assess the role of CRMP2 in periventricular leukomalacia (PVL). We developed a PVL model using 3-day-old rats to investigate the expression and phosphorylation of CRMP2 in the newborn brain. Hypoxia–ischemia was applied by unilateral carotid ligation followed by exposure to 5% oxygen for 30min. Pathological changes were evaluated from 0h to 21d post-HI, and white matter damage including severe necrosis, white matter rarefaction and lateral ventricle dilatation were found. In the PVL model astrogliosis and axonal damage were detected in the injured white matter by immunohistochemistry at 48–168h post-HI, and delayed myelination was verified by Western blotting after 21-day post-HI. We confirmed that this model showed neuropathological features of PVL. Next, significant changes of CRMP2 were observed in the brain of the PVL model. Western blotting and immunohistochemistry showed that cleavage and hypo-phosphorylation of CRMP2 occurred after 48h post-HI in the PVL brain. Our results suggest that cleaved CRMP2 could represent hypo-phosphorylated-CRMP2 and HI could induce activation of CRMP2 in the PVL brain. The activated CRMP2 may play an important role in neuronal plasticity in PVL. Our findings suggest that future treatment strategies of PVL should target the phosphorylation mechanism of CRMP2. [Copyright &y& Elsevier]

Spontaneous correlated neuronal activity during early development spreads like a wave by recruiting a large number of neurons, and is considered to play a fundamental role in neural development. One important and as yet unresolved question is where the activity originates, especially at the earliest stage of wave expression. In other words, which part of the brain differentiates first as a source of the correlated activity, and how does it change as development proceeds? We assessed this issue by examining the spatiotemporal patterns of the depolarization wave, the optically identified primordial correlated activity, using the optical imaging technique with voltage-sensitive dyes. We surveyed the region responsible for the induction of the evoked and spontaneous depolarization waves in chick embryos, and traced its developmental changes. The results showed that the wave initially originated in a restricted area near the obex and was generated by multiple regions at later stages. We suggest that the upper cervical cord/lower medulla near the obex is the kernel that differentiates first as the source of the correlated activity, and that regional and temporal differences in neuronal excitability might underlie the developmental profile of wave generation in early chick embryos. [ABSTRACT FROM AUTHOR]

1. Multiple-site optical recording with the voltage-sensitive absorption dye NK2761 was used to investigate functiogenesis of the neural circuits in the embryonic central nervous system (CNS). In the present review, we emphasize the functional development of glossopharyngeal nerve (N. IX)- and vagus nerve (N. X)-related neural circuits in the embryonic chick brainstem. 2. In the nucleus tractus solitarius (NTS), glutamatergic excitatory post-synaptic potentials (EPSPs) evoked by stimulation of N. IX/N. X were first detected at Embryonic Day (E) 7, when morphological differentiation of pre- and post-synaptic neurons is incomplete. The removal of extracellular Mg2+ elicited small EPSPs at E6, suggesting that synaptic function mediated by N-methyl-d-aspartate (NMDA) receptors (glutamatergic receptors) is latently generated 1 day before the expression of glutamatergic EPSPs. In the nucleus of the glossopharyngeal nerve and the dorsal motor nucleus of the vagus nerve, action potentials were detected from E3.5 and E4, respectively. Similar chronological sequences were observed in the rat embryo and in other cranial nerve-related nuclei and were used to construct basic profiles of embryonic EPSPs in the vertebrate CNS. 3. We further investigated the development of the secondary synaptic pathways from the NTS to higher centres and found that neuronal circuits from the NTS are already generated when the primary afferents form functional synapses with NTS neurons. 4. A widely spreading depolarization wave was found in the embryonic CNS. This wave was triggered by multisensory inputs and spontaneous activities and was suggested to be mediated by a dual network of chemical synapses and gap junctions. It was also accompanied by a Ca2+ wave, indicating its nurturing effects on CNS development. [ABSTRACT FROM AUTHOR]

Abstract: Using voltage-sensitive dye recording, we traced the ontogenetic expression of neural excitability related to the glossopharyngeal nerve (N. IX) and the vagus nerve (N. X) in the embryonic chick brainstem. At the 3.5-day embryonic stage, by averaging optical signals, we succeeded in recording very small action potential-related optical responses (ΔI/I <10−4) induced by N. IX stimulation in the nucleus of the glossopharyngeal nerve (motor nucleus). This suggests that glossopharyngeal excitability in the motor nucleus is first generated no later than this developmental stage. On the other hand, action potential-related optical responses induced by N. X stimulation were first detected at the 4-day embryonic stage. Comparison with morphogenesis indicated that glossopharyngeal and vagal motoneurons acquire their excitability and send their axons to the periphery soon after they leave the proliferative pool. [Copyright &y& Elsevier]

Membrane-embedded voltage-sensor domains in voltage-dependent potassium channels (Kv channels) contain an impressive number of charged residues. How can such highly charged protein domains be efficiently inserted into biological membranes? In the plant Kv channel KAT1, the S2. S3, and 54 transmembrane helices insert cooperatively, because the S3, 54, and 53-54 segments do not have any membrane insertion ability by themselves. Here we show that, in the Drosophila Shaker Kv channel, which has a more hydrophobic S3 helix than KAT1, 53 can both insert into the membrane by itself and mediate the insertion of the S3-54 segment in the absence of 52. An engineered KAT1 53-54 segment in which the hydrophobicity of 53 was increased or where 53 was replaced by Shaker 53 behaves as Shaker S3-54. Electrostatic interactions among charged residues in 52, S3, and 54, including the salt bridges between E283 or E293 in S2 and R368 in S4, are required for fully efficient membrane insertion of the Shaker voltage-sensor domain. These results suggest that cooperative insertion of the voltage-sensor transmembrane helices is a property common to Kv channels and that the degree of cooperativity depends on a balance between electrostatic and hydrophobic forces. [ABSTRACT FROM AUTHOR]

Abstract: Terrestrial palynomorph assemblages were found from the Lower Cretaceous Miyako Group in the Tanohata area, northeast Japan, and the Tetori Group in the Kuzuryu area, central Japan. There is no obvious difference between the assemblages from these two localities, although these localities have been regarded as belonging to different paleophytogeographic provinces in East Asia on the basis of the plant megafossils. This result is consistent with the suggestion by the recent reconsideration on the megafossil flora from the Tetori Group that the paleophytogeographic provincialism might have weakened during middle to late Early Cretaceous time. The terrestrial palynomorph assemblage from the Miyako Group has characters of both of Cerebropollenites and Schizaeoisporites palynofloras, which were proposed by previous studies, in the Northern Hemisphere during the Early Cretaceous time. This suggests an overlooked transitional palynofloral province between the Cerebropollenites and Schizaeoisporites palynofloral provinces in continental margin of Eurasia during that time. [Copyright &y& Elsevier]

During development, correlated neuronal activity plays an important role in the establishment of the central nervous system (CNS). We have previously reported that a widely propagating correlated neuronal activity, termed the depolarization wave, is evoked by various sensory inputs. A remarkable feature of the depolarization wave is that it spreads broadly through the brain and spinal cord. In the present study, we examined whether the depolarization wave occurs spontaneously in the embryonic rat CNS and, if so, where it originates. In E15–16 rat embryos, spontaneous optically-revealed signals appeared in association with the rhythmic discharges of cranial motoneurons and propagated widely with similar characteristics to the evoked depolarization wave. At E15, the spontaneous wave mostly originated in the cervical to upper lumbar cords. At E16, the wave was predominantly generated in the lumbosacral cord although a wave associated with the second oscillatory burst was initiated in the rostral cord. At E16, a few waves also originated in the rostral ventrolateral medulla and the dorsomedial pons. When the influence of the caudal cord was removed by transecting the spinal cord, the contribution of the medulla and pons became more significant. These results show that the depolarization wave can be triggered by the spontaneous activity of multiple neuronal populations which are distributed widely from the pons to the lumbosacral cord, although the spinal cord usually plays a predominant role. This network possibly works as a self-distributing system that maintains the incidence and complicated patterns of the correlated activity in the developing CNS. [ABSTRACT FROM AUTHOR]

Abstract: We examined the intracellular Ca2+ ([Ca2+]i) elevation evoked by GABA in an 8-day embryonic chick brainstem slice using a Ca imaging technique with Ca green-1 AM. When small quantities of GABA were pressure-ejected on the surface of the slice, the [Ca2+]i elevation was clearly detected. The GABA-induced [Ca2+]i elevation was eliminated in a Ca2+-free solution, whereas the previously reported GABA-induced light-scattering change was independent of extracellular Ca2+. Although, micro-application of glycine or glutamate also induced [Ca2+]i elevation, these changes were smaller than that by GABA. These results suggest that the GABA-induced [Ca2+]i elevation is due to Ca2+ entry resulting from membrane depolarization and may play an important role in the development of the central nervous system (CNS). [Copyright &y& Elsevier]

Objective: To evaluate the expression patterns of heat shock transcription factor, Y chromosome (HSFY), in the testes showing deteriorated spermatogenesis. Design: Prospective study. Setting: University hospital, its branch hospital, and academic laboratory. Patient(s): Men undergoing testicular biopsy for the investigation of infertility and men undergoing orchiectomy for testicular cancer. Intervention(s): After pathologic evaluation, specimens were subdivided into three groups: normal spermatogenesis (n = 8), maturation arrest (n = 5), and Sertoli cell–only syndrome (n = 4). Immunostaining and Western blotting techniques determined the expression of HSFY. Main Outcome Measure(s): Expression of HSFY in testes. Result(s): Western blotting data revealed HSFY in the testicular tissues with normal spermatogenesis, maturation arrest, and Sertoli cell–only syndrome, but the amount of the protein in the maturation arrest and Sertoli cell–only syndrome samples was altered. The immunohistochemical data demonstrated that HSFY was expressed in spermatogenic cells and Sertoli cells in all specimens. However, the expression of HSFY was low or absent in spermatogenic cells of maturation arrest specimens, and the ratio of HSFY expressed in Sertoli cells was different in the specimens with maturation arrest and with Sertoli cell–only syndrome. Conclusion(s): Altered expression of the HSFY in the testis showing deteriorated spermatogenesis may be associated with alteration of spermatogenic cell differentiation. [Copyright &y& Elsevier]

Abstract: Multiple-site optical recording with a fast voltage-sensitive dye, absorption dye NK2761, was used to study the developmental organization of functional synaptic networks in the vagal pathway. Glutamatergic excitatory postsynaptic potentials (EPSPs) evoked by vagus nerve stimulation was first detected from the nucleus of the tractus solitarius (NTS) at embryonic day 7 (E7) in chick embryos and E15 in rat embryos, when morphological differentiation of pre- and postsynaptic neurons is incomplete. When extracellular Mg2+ was removed, small EPSPs were elicited at E6 in chick embryos and E14 in rat embryos. These results suggest that synaptic function mediated by N-methyl-d-aspartate (NMDA) receptors is latently generated 1 day before the expression of glutamatergic EPSP. Functional synapses related to the glossophyaryngeal nerve appear to be generated at the same time as the vagus nerve, but their spatial distribution was different from that of the vagus nerve. We further investigated the development of second synaptic pathways from the NTS to higher centers, and found that neuronal circuits from the NTS are already generated when the primary afferents form functional synapses with NTS neurons. [Copyright &y& Elsevier]

Abstract: Shell of the adult hermit crab has some important roles for its fitness. In the same time, the shell size often limits the body growth of its owner. To grow the body size larger, the individual must change the shell to another larger shell. If the individual cannot get another larger one, the individual has to suppress the body size growth as the occupied shell size allows. Growth suppression would result in the lower fitness. With a simple mathematical model, we consider the criterion about whether the individual should try to change the shell or not in order to get the higher fitness. We show that the optimality of a shell change behavior has a relation with the body size and the season length for the shell change. They also affect the optimal timing for the shell change. It is implied that the probability of the success in a shell change and the cost for the shell change behavior do not affect the optimal timing for the shell change at all but significantly do the optimality of the behavioral choice. [Copyright &y& Elsevier]

The membrane topology of the Arabidopsis thaliana Na+/H+ exchanger isoform 1 (AtNHX1) was investigated by examining the topogenic function of transmembrane (TM) segments using a cell-free system. Even though the signal peptide found in the human Na+/H+ exchanger (NHE) family is missing, the N-terminal hydrophobic segment was efficiently inserted into the membrane and had an N-terminus lumen topology depending on the next TM segment. The two N-terminal TM segments had the same topology as those of TM2 and TM3 of human NHE1. In contrast, TM2 and TM3 of human NHE1 did not acquire the correct topology when the signal peptide (denoted as TM1) was deleted. Furthermore, there were three hydrophobic segments with the same topogenic properties as the TM9-H10-TM10 segments of human NHE1, which has one lumenal loop (H10) and two flanking TM segments (TM9 and TM10). These data indicate that the plant NHX isoforms can form the common membrane topology proposed for the human NHE family, even though it does not have a signal peptide. [ABSTRACT FROM PUBLISHER]

Multiple-site optical recording of neural activity was performed in the nucleus of the tractus solitarius (NTS) of the chick embryo with stimulation of the glossopharyngeal nerve (N. IX) and vagus nerve (N. X). We measured the amplitudes of the optical signals related to glutamate-mediated excitatory postsynaptic responses, and calculated the ratio of the signal evoked by simultaneous N. IX/N. X stimulation to the signal obtained after mathematical summation of the individual N. IX and N. X responses. The ratio was significantly lower than 100% in the rostral region of the NTS, in which postsynaptic responses were elicited by both N. IX and N. X stimulations. This result means that there is a convergence of visceral inputs via the N. IX and N. X in the embryonic chick NTS. The existence of the convergence suggests that the NTS performs complex integration of information from multiple sensory inputs from the early stages of embryogenesis. [Copyright &y& Elsevier]

Multispanning membrane proteins are synthesized by membrane-bound ribosomes and integrated into the endoplasmic reticulum membrane cotranslationally. To uncover the topogenic process of membrane loop, of which both ends are in the same side of the membrane, we examined topogenesis of a relatively hydrophobic lumenal loop segment (H10 segment) between TM9 and TM10 of human Na+/H+ exchanger isoform 1 using an in vitro expression system. The H10 segment was translocated through the membrane. Any potential sites created within the H10 segment were not glycosylated. Just after TM9, there are potential glycosylation and signal peptidase processing sites. When the reporter domain of prolactin was fused at the position preceding the H10 segment, these sites were modified by the enzymes, while they were not modified in the original molecule. Thus, we concluded that the H10 segment was translocated through the membrane and directly inserted into the membrane and that its membrane insertion caused sequestration of the preceding processing and glycosylation sites from the lumenal modifying enzymes. This topogenic process shows clear contrast to that of pore loops of K+ channels, which are once exposed in the lumen and accessible to glycosylation enzyme. [Copyright &y& Elsevier]

To examine whether there are any differences in functional organization between the glossopharyngeal nerve (N. IX)- and vagus nerve (N. X)-projecting areas in the nucleus of the tractus solitarius (NTS), we performed optical recording of neural responses evoked by N. IX stimulation in 5- to 9-day-old embryonic chick brain stem preparations and compared the results with those in our previous studies concerning the N. X-related NTS. First, we investigated DL-2-amino-5-phosphonovaleric acid (APV)/Mg2+ sensitivity of the glutamatergic excitatory postsynaptic potentials (EPSPs) in the N. IX-related NTS. In 7- to 9-day-old preparations, we found regional differences in the degree of both the APV-induced reduction and Mg2+-free-induced enhancement of the EPSPs. We constructed developmental maps of spatial patterns of the APV- and Mg2+-sensitive components and showed that functional expression of the N-methyl-D-aspartate (NMDA) receptor dynamically changed during development. Second, we studied initial expression of synaptic functions in the N. IX-related NTS. In 6-day-old preparations, although action potentials alone were usually detected in normal Ringer solution, small EPSPs were elicited in a Mg2+-free solution. This result suggests that the NMDA receptor-mediated synaptic function is latently generated in the N. IX-related NTS at the 6-day-old embryonic stage and that external Mg2+ regulates the onset of synaptic functions. Developmental patterns of APV/Mg2+ sensitivity and the stage of initial expression of the glossopharyngeal EPSP were similar to those of the N. X, suggesting that the developmental sequence of the synaptic function in the NTS is the same for the N. IX- and N. X-related NTS. [ABSTRACT FROM AUTHOR]

We examined the functional organization of the rat trigeminal nuclear complex and its developmental dynamics using a multiple-site optical recording technique. Brainstem preparations were dissected from embryonic day 12 (E12)-E16 rat embryos, and stimulation was applied individually to the three branches of the trigeminal nerve (V1-V3). The action potential activity of presynaptic fibers was detected from E13, and the glutamate-mediated postsynaptic response was significantly observed from E15 on. At E14, the evoked signals usually consisted of only the action potential-related fast component. However, when extracellular Mg2+ was removed, a significant DL-2-amino-5-phosphonovaleric acid-sensitive slow component appeared. These results suggest that postsynaptic function mediated by NMDA receptors is latently generated as early as E14. The response area of the three branches of the trigeminal nerve showed some functional somatotopic organization, with the ophthalmic (V1) nerve area medially located and the mandibular (V3) nerve area laterally located. The center of the trigeminal nuclear complex in which the activity of neurons and synaptic function was greatest shifted caudally with development, suggesting that the functional architecture of the trigeminal nuclear complex is not fixed but changes dynamically during embryogenesis. By electron microscopy, we could not observe clear correlations between functional data and morphological information; when we surveyed E16 preparations, we could not identify typical synaptic structures between the 1,1′-dioctyldecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate-labeled trigeminal nerve terminals and the neurons in the trigeminal nuclear complex. This implies that postsynaptic function in the trigeminal nuclear complex is generated before the appearance of the morphological structure of conventional synapses. [ABSTRACT FROM AUTHOR]

Potassium channels, which control the passage of K[sup +] across cell membranes, have two transmembrane segments, M1 and M2, separated by a hydrophobic P region containing a highly conserved signature sequence. Here we analyzed the membrane topogenesis characteristics of the M1, M2, and P regions in two animal and bacterial two-transmembrane segment-type K[sup +] channels, Kir 2.1 and KcsA, using an in vitro translation and translocation system. In contrast to the equivalent transmembrane segment, S5, in the voltage-dependent K[sup +] channel, KAT1, the MI segment in KcsA, was found to have a strong type H signal-anchor function, which favors the N[sub cyt]/C[sub exo] topology. The N-terminal cytoplasmic region was required for efficient, correctly orientated integration of MI in Kit 2.1. Analysis of N-terminal modification by in vitro metabolic labeling showed that the N terminus in Kir 2.1 was acetylated. The hydrophobic P region showed no topogenic function, allowing it to form a loop, but not a transmembrane structure in the membrane; this region was transiently exposed in the endoplasmic reticulum lumen during the membrane integration process. M2 was found to possess a stop-transfer function and a type I signal-anchor function, enabling it to span the membrane. The C-terminal cytoplasmic region in KcsA was found to affect the efficiency with which the M2 achieved their final structure. Comparative topogenesis studies of Kir 2.1 and KcsA allowed quantification of the relative contributions of each segment and the cytoplasmic regions to the membrane topology of these two proteins. The membrane topogenesis of the pore-forming structure is discussed using results for Kir 2.1, KcsA, and KAT1. [ABSTRACT FROM AUTHOR]

Discusses the molecular dissection of the contribution of negatively and positively charged residues in S2, S3 and S4 to the final membrane topology of the voltage sensor in the K[sup +] channel, KAT1. Use of in vitro translation and translocation experiments; Indication that Asp[sup 95] assists in the membrane insertion S3-S4.

Identifies the negative residues of the second transmembrane segment in the plant voltage-dependent K[sup+] channel, KAT1, which involves the function of voltage sensing. Failure of point mutations at D95 and D105 to complement the K[sup+] uptake deficient properties of the mutant yeast; Role played by both residues in the membrane topogenesis of the voltage sensor KAT1.

Describes the integration of Shaker-type K[sup +] channel, KAT1 into the endoplasmic reticulum (ER) membrane. Effect of the inserted glycosylation sequence; Accounts on the topogenesis mechanism transmembrane segments; Accession of oligosaccharyl transferase in the ER membrane.

Regular health checkups are important for mothers and newborns to detect health problems at an early stage; however, this is often difficult in resource-limited settings. Therefore, the portable health clinic (PHC) for maternal and child health (MCH), a telemedicine health checkup system, was introduced as an intervention study in a rural area in Bangladesh. The aim of this research project was to report findings that we had observed at a mid-point of the intervention period. This was an intervention study conducted in Shariatpur, Bangladesh. The study population included pregnant/parturient women aged 15–49 years and their newborns. With the help of the newly created PHC for MCH, health workers, with a set of sensor devices in an attaché case, visited mothers and newborns at home to examine their health status. Their health status was triaged into four categories using a data management application, and in cases of affected or emergent health status, they were placed on remote video consultation with a doctor. In total, 94 women were included in the PHC for MCH intervention. The rate of participants who received antenatal care at least four times or postnatal care at least once increased (from 29% to 51%, and from 27% to 78%, respectively) compared with before introducing PHC for MCH. Using the PHC for MCH, we detected health problems in pregnant/parturient women; a relatively high percentage had anemia (45–54%) and/or abnormal pulse rate (20–40%). Moreover, after introducing the PHC for MCH, more than 40% of women who received multiple antenatal care or postnatal care checkups improved their health status. The PHC for MCH could be an effective system to improve the health of mothers and newborns by increasing the availability of care. In the future, this system is expected to be used as a primary resource for maternity healthcare, not only in rural areas but also in other social environments. [ABSTRACT FROM AUTHOR]