A CatSper-Uninvolved Mechanism to Induce Forward Sperm Motility in the Internal Fertilization.

Sperm-specific cation channel (CatSper), sperm-specific Na + /H + exchanger (sNHE), and soluble adenylyl cyclase (sAC) are necessary in the signaling pathways to control sperm motility in many animals, whereas some animals have lost some or all of them. In the present study, we examined CatSper-uninvolved signaling for vigorous undulation of the undulating membrane that is attached to the sperm tail and gives thrust for forward motility in the internally fertilizing newt Cynops pyrrhogaster. Reverse-transcription PCR failed to detect sNHE in the newt sperm. However, the pH of sperm cytoplasm was raised under a high extracellular pH equivalent to that of egg jelly, where sperm motility is initiated by sperm motility-initiating substance (SMIS). Carbonic anhydrase XII/ XVI and SLC4A4/8 were suggested to be present in the sperm, and transported bicarbonates raised the intracellular pH. In egg jelly extract that contained SMIS, the anion transporter inhibitor DIDS weakened the undulation of the undulating membrane, while bicarbonates enhanced it. The cyclic AMP concentration was found to increase in sperm cytoplasm in the egg-jelly extract. An inhibitor of sAC (KH7) weakened the undulation of the undulating membrane, and dibutyryl cyclic AMP blocked the inhibitory effect. Inhibitor of transmembrane AC (DDA) limitedly affected the undulation. The undulation was weakened by an inhibitor of protein kinase A (H89), and by an inhibitor of transient receptor potential (TRP) channels (RN1747). Our results support the conclusions that the high pH of the egg jelly triggers a signaling pathway through sAC, PKA, and TRP channels, and coacts with SMIS to induce forward sperm motility.

Sperm storage influences the potential for spontaneous acrosome reaction of the sperm in the newt Cynops pyrrhogaster.

Sperm storage is supposed to influence sperm quality, although the details remain unclear. In the present study, we found that sperm stored in a sperm storage site, the vas deferens of Cynops pyrrhogaster, spontaneously undergo acrosome reaction following incubation in Steinberg's salt solution (ST). Percentages of acrosome-reacted sperm increased time-dependently to about 60% in 24 hr. The concentration of cyclic adenosine monophosphate (cAMP) was elevated after incubating sperm in ST, while dibutylyl cAMP induced an acrosome reaction. Chelating of extracellular Ca2+ suppressed the dibutylyl cAMP-induced acrosome reaction as well as spontaneous acrosome reaction in ST. These results suggest that cAMP elevation driven by Ca2+ influx can be a cue for spontaneous acrosome reaction. Relatively low Ca2+ concentration and pH in the vas deferens were sufficient to suppress spontaneous acrosome reaction within 1 hr. In addition, the cysteine rich secretory protein 2 gene was expressed in the vas deferens, indicating that it may be involved in the continuous suppression of spontaneous acrosome reaction. Sperm that underwent spontaneous acrosome reaction in ST was significantly increased when stored in the vas deferens for longer periods, or by males experiencing temperatures in excess of 12°C during hibernation conditions. Percentages of the spontaneously acrosome-reacted sperm were found to differ among males even though they were of identical genetic background. Taken together, C. pyrrhogaster sperm possess the potential for spontaneous acrosome reaction that does not become obvious in the vas deferens, unless promoted in correlation with sperm storage.

Distinct Ca2+ channels maintain a high motility state of the sperm that may be needed for penetration of egg jelly of the newt, Cynops pyrrhogaster.

Activation state of sperm motility named "hyperactivation" enables mammalian sperm to progress through the oviductal matrix, although a similar state of sperm motility is unknown in non-mammalian vertebrates at fertilization. Here, we found a high motility state of the sperm in the newt Cynops pyrrhogaster. It was predominantly caused in egg jelly extract (JE) and characterized by a high wave velocity of the undulating membrane (UM) that was significantly higher at the posterior midpiece. An insemination assay suggested that the high motility state might be needed for sperm to penetrate the egg jelly, which is the accumulated oviductal matrix. Specific characteristics of the high motility state were completely abrogated by a high concentration of verapamil, which blocks the L-type and T-type voltage-dependent Ca(2+) channels (VDCCs). Mibefradil, a dominant blocker of T-type VDCCs, suppressed the wave of the UM at the posterior midpiece with separate wave propagation from both the anterior midpiece and the posterior principal piece. In addition, nitrendipine, a dominant L-type VDCC blocker, weakened the wave of the UM, especially in the anterior midpiece. Live Ca(2+) imaging showed that, compared with the intact sperm in the JE, the relative intracellular Ca(2+) level changed especially in the anterior and posterior ends of the midpiece of the blocker-treated sperm. These suggest that different types of Ca(2+) channels mediate the intracellular Ca(2+) level predominantly in the anterior and posterior ends of the midpiece to maintain the high motility state of the newt sperm.

[Development and application of microbiology data monitoring system (Akita-ReNICS) for networking hospitals in Akita Prefecture].

Networking among regional hospitals recently became required for infection control activities in Japan. In Akita prefecture, we developed a microbiology data warehouse for networking hospital laboratories in 2010. Designated users can easily obtain microbiology data and compare it with other participating hospitals using this system. For example, users are able to compare the MRSA/MSSA ratio in Staphylococcus aureus with other participating hospitals in Akita-ReNICS. The effectiveness and assignment of this system were shown by a questionnaire administrated in 2013. In this paper, we clarify the current state and assignment of Akita-ReNICS and consider its effective application for better infection control activities.

[Development of a microbiology data warehouse (Akita-ReNICS) for networking hospitals in a medical region].

The active involvement of hospital laboratory in surveillance is crucial to the success of nosocomial infection control. The recent dramatic increase of antimicrobial-resistant organisms and their spread into the community suggest that the infection control strategy of independent medical institutions is insufficient. To share the clinical data and surveillance in our local medical region, we developed a microbiology data warehouse for networking hospital laboratories in Akita prefecture. This system, named Akita-ReNICS, is an easy-to-use information management system designed to compare, track, and report the occurrence of antimicrobial-resistant organisms. Participating laboratories routinely transfer their coded and formatted microbiology data to ReNICS server located at Akita University Hospital from their health care system's clinical computer applications over the internet. We established the system to automate the statistical processes, so that the participants can access the server to monitor graphical data in the manner they prefer, using their own computer's browser. Furthermore, our system also provides the documents server, microbiology and antimicrobiotic database, and space for long-term storage of microbiological samples. Akita-ReNICS could be a next generation network for quality improvement of infection control.

[Case in which the unilateral intraocular pressure before hemodialysis was higher than the day following hemodialysis].

PURPOSE: To report a case in which the unilateral intraocular pressure (IOP) before hemodialysis was higher than the day following hemodialysis. CASE: A 59-year-old woman had been followed with diabetic retinopathy in a local eye clinic and was referred to our hospital for a vitreous hemorrhage in the right eye in June 2002. She started hemodialysis for renal failure due to diabetic nephropathy in July 2002; vitreous and cataract surgeries were performed in September 2002. In May 2004, she underwent surgery in the left eye. The IOP in the left eye increased to the high 20s in August 2005. The visual acuity in the right eye was 0.7 and 0.3 in the left eye. The angles were open bilaterally. Before hemodialysis, the IOP in the left eye was significantly higher than that in the days following hemodialysis. There was no significant change in the right eye between before and after hemodialysis. Following trabeculectomy performed in the left eye in January 2007, the IOP in the left eye stabilized in the low teens. CONCLUSION: In some cases, the IOP can vary between before and the days following dialysis. It is important to check the IOP at these time points.

[Clinical evaluation after an introduction of our manual for antibiotic use during perioperative period and a notification policy of use of the antibiotics for MRSA].

Abuse of antibiotics results in unfavorable consequences in healthcare associated infection control, such as emergence of multiple-resistant bacteria, and increased medical cost and nosocomial infection. In 2003, we had an outbreak of Methicillin-resistant Staphylococcus aureus (MRSA) in a surgical ward of Akita University Hospital. With the aim of preventing inappropriate use of antibiotics during perioperative period, a guideline for usage of antibiotics, which is made by surgeons themselves, was introduced, and a notification policy of the antibiotic use for MRSA was also applied in 2004. In this study, we evaluated the influence of the antibiotic restriction on changes in antibiotics consumption, prevalence of multiple-drug-resistant pathogens, and their susceptibility to antibiotics. The notification policy reduced the cost and amount of antibiotics and the prevalence of multiple-drug-resistant pathogens, such as MRSA and Pseudomonas aeruginosa. The susceptibility of bacteria, such as Staphylococcus aureus, MRSA, Serratia marcescens and Pseudomonas aeruginosa, to antibiotics showed a remarkable improvement. The introduction of notification policy for usage of antibiotics has a favorable influence on the infection control in hospitals and re education of doctors.

EG2 expressed by eosinophils as a clinically useful indicator of asthma.

The monitoring of airway inflammation is mandatory for the improved control of bronchial asthma. We previously reported that intracellular EG2 levels of eosinophils, a marker of bronchial asthma increased in asthma patients. In this study, we hypothesized that eosinophil EG2(+) expression increases during airway inflammation in asthmatic individuals. Eosinophil EG2(+) and percentage eosinophil EG2(+) with whole blood flow cytometry, eosinophil counts, serum total IgE, serum eosinophil cationic protein, eosinophil-derived neurotoxin, and percent of forced expiratory volume in 1 second/force vital capacity (FEV(1)/FVC) were measured in 33 asthmatic patients and 22 healthy volunteers. The relationships between these markers were evaluated. Comparisons were made on EG2(+) expression between attack and asymptomatic periods in six asthmatic patients. EG2(+) expression was significantly greater in the asthmatic patients than in healthy subjects. Furthermore, the EG2(+) expression showed a significant increase during attacks. EG2(+) expression inversely correlated with the FEV(1)/FVC. These results suggest that EG2(+) expression may be a useful clinical marker of airway inflammation in asthma.

Co-existence of Mycobacterium tuberculosis and Mycobacterium intracellulare in one sputum sample.

An 81-year-old man was admitted to the hospital with a severe sore throat and a low grade fever. A chest radiograph showed bilateral diffuse reticulonodular shadows. By fluorescent stain for mycobacteria, his sputum smear showed acid-fast bacteria. The initial polymerase chain reaction (PCR) of his sputum revealed Mycobacterium intracellulare (M. intracellulare), but not Mycobacterium tuberculosis (M. tuberculosis). However, a repeat PCR was performed because M. tuberculosis could not be ruled out due to his clinical symptoms and chest imaging. The second PCR detected both M. intracellulare and M. tuberculosis. From the standpoint of infection control, this case illustrates the possibility that M. tuberculosis could be a threat if a second PCR is not done. While PCR is a useful exam for diagnosing M. tuberculosis, it can produce false negative results. Therefore, for diagnosing tuberculosis, particularly in a case such as the present case, a second PCR, which is not normally necessary, should be done.