Intestinal homeostasis and inflammation: Gut microbiota at the crossroads of pancreas-intestinal barrier axis.

The pancreas contains exocrine glands, which release enzymes (e.g., amylase, trypsin, and lipase) that are important for digestion and islets, which produce hormones. Digestive enzymes and hormones are secreted from the pancreas into the duodenum and bloodstream, respectively. Growing evidence suggests that the roles of the pancreas extend to not only the secretion of digestive enzymes and hormones but also to the regulation of intestinal homeostasis and inflammation (e.g., mucosal defense to pathogens and pathobionts). Organ crosstalk between the pancreas and intestine is linked to a range of physiological, immunological, and pathological activities, such as the regulation of the gut microbiota by the pancreatic proteins and lipids, the retroaction of the gut microbiota on the pancreas, the relationship between inflammatory bowel disease, and pancreatic diseases. We herein discuss the current understanding of the pancreas-intestinal barrier axis and the control of commensal bacteria in intestinal inflammation.

A modified GnRH antagonist method in combination with letrozole, cabergoline, and GnRH antagonist for PCOS: Safe and effective ovarian stimulation to treat PCOS and prevent OHSS.

PURPOSE: To analyze the therapeutic efficacy of a modified controlled ovarian stimulation (COS) protocol for polycystic ovary syndrome (PCOS) that does not cause ovarian hyperstimulation syndrome (OHSS) while maintaining oocyte quality. METHOD: This study is a retrospective cohort study of reproductive medicine at St. Mother Clinic. We analyzed ART clinical outcomes, embryonic development, and hormone levels in 175 PCOS patients treated with four COS (GnRH agonist based long protocol, Group A; GnRH antagonist protocol with HCG trigger, Group B; GnRH antagonist protocol with GnRH agonist trigger, Group C, and the modified COS group) between 2010 and 2021. RESULTS: Of 175 patients with PCOS, 45 and 130 patients underwent 47 and 136 oocyte retrieval cycles, 75 and 250 embryo transfer cycles with the modified COS, and with conventional methods, respectively. The cumulative pregnancy rate at one trial was a significantly higher result than in Group A and higher than in Groups B and C (cumulative pregnancy rate at one trial of Group A, B, C, and modified COS: 40.0%, 54.5%, 56.3%, and 72.3%, respectively). With this method, not clinically problematic OHSS and higher clinical outcomes than in conventional methods were observed. CONCLUSION: This modified COS can significantly improve clinical outcomes and eliminate OHSS.

Modified permeable cryoprotectant-free vitrification method for three or fewer ejaculated spermatozoa from cryptozoospermic men and 7-year follow-up study of 14 children born from this method.

STUDY QUESTION: What technique can be used to successfully cryopreserve three or fewer ejaculated spermatozoa from cryptozoospermic men and is the physical and cognitive development of children born after this technique normal? SUMMARY ANSWER: The modified cryopreservation method for three or fewer human spermatozoa from cryptozoospermic men showed a recovery rate above 95% and a survival rate just under 90%, and the physical and cognitive abilities of the children born after ICSI were comparable to those born after natural conception. WHAT IS KNOWN ALREADY: Clinical outcomes of ICSI using cryptozoospermic men's ejaculated spermatozoa are considered to be inferior to that using testicular spermatozoa from microsurgical testicular sperm extraction (Micro-TESE), possibly because the DNA fragmentation rate is higher in ejaculated spermatozoa than in testicular spermatozoa from Micro-TESE. STUDY DESIGN, SIZE, DURATION: Evaluation of the efficiency of cryopreservation of three or fewer spermatozoa was conducted retrospectively at St. Mother Clinic. The physical and cognitive development of children born after this method was studied between 2011 and 2018. PARTICIPANTS/MATERIALS, SETTING, METHODS: This study included 28 cryptozoospermic men who had three or fewer morphologically normal and motile spermatozoa in their ejaculate after centrifugation and who preferred using cryopreserved spermatozoa to Micro-TESE. Control subjects were 31 cryptozoospermic patients using fresh spermatozoa from their ejaculates and 20 non-obstructive azoospermic patients with fewer than 10 spermatozoa obtained by TESE and vitrified. Clinical outcomes among three groups, vitrified spermatozoa from the ejaculate, fresh spermatozoa from the ejaculate and vitrified spermatozoa from the testis, were statistically analysed. For the 7-year follow up study of the 14 children born after ICSI using the ejaculated vitrified spermatozoa, the Japanese government-issued Boshi Kenko Techo (Mother-Child Handbook) and Kinder Infant Development Scale (KIDS scale) were used to determine whether their physical and cognitive development was comparable to that of naturally conceived children. MAIN RESULTS AND THE ROLE OF CHANCE: Recovery and survival rates were 97.8% (510/521) and 87.1% (444/510) for vitrified spermatozoa from the ejaculate and 92.7% (152/164) and 60.5% (92/152) for vitrified spermatozoa from the testis. Clinical pregnancies (%), miscarriages (%) and live birth rates (%), respectively, among the three groups were as follows: vitrified spermatozoa from the ejaculate: 15(25.0), 2(13.3), 13(21.7); fresh spermatozoa from the ejaculate: 26(24.3), 5(19.2), 20(18.7); and vitrified spermatozoa from the testis: 3(16.7), 0(0.0), 3(16.7). Among the groups, there were no statistically significant differences except for the sperm survival rate and the oocyte fertilisation rate, which were lower for vitrified spermatozoa from the testis compared with vitrified spermatozoa from the ejaculate. The 7-year follow-up study showed that the physical and cognitive development of 14 children born after ICSI using vitrified ejaculated spermatozoa from the ejaculate was comparable to that of naturally conceived children. LIMITATIONS, REASONS FOR CAUTIONS: The maximum number of spermatozoa to which this method can be applied successfully is about 10. When the number of aspirated spermatozoa is over 10, some of them change direction after colliding with each other inside the aspiration pipette and reach the mineral oil, and once this happens, they cannot be expelled out of the pipette. Even though we did not find evidence of DNA fragmentation, further studies with larger participant numbers and longer time periods are necessary. WIDER IMPLICATIONS OF THE FINDINGS: This technique is very useful for the cryopreservation of very small numbers of testicular spermatozoa (fewer than 10) in order to avoid or reduce Micro-TESE interventions. STUDY FUNDING/COMPETING INTEREST(S): No external funding was received to undertake this study. There are no competing interests. TRIAL REGISTRATION NUMBER: N/A.

[Awareness of the term "frailty" and its correlates among older adults living in a metropolitan area].

Objectives　This study aimed to examine the awareness of the term "frailty" and its correlates among older adults living in a Japanese metropolitan area.Methods　We used baseline and 2-year follow-up data from a community-wide intervention on preventing frailty in Ota City, Tokyo. In July 2016, we conducted a self-administered questionnaire survey via mail to investigate the lifestyle and health status of 15,500 non-disabled residents aged 65-84 years from all 18 districts. In July 2018, we investigated the awareness of the term "frailty" in the same sample using a different questionnaire, and 10,228 people (4,977 men and 5,251 women) were included in the analyses. In addition, 9,069 people (4,347 men and 4,722 women) who responded to both 2016 and 2018 surveys were analyzed for correlates of awareness of the term "frailty." We categorized the responses "I know the meaning" or "I have heard of it but do not know the meaning" as awareness of the term. Decision tree and multilevel Poisson regression analyses were performed to examine the association of the following with awareness of "frailty": age, marital status, living situation, education, equivalent income, body mass index, number of chronic diseases, alcohol consumption, smoking status, Dietary Variety Score (DVS), and presence or absence of lower back and knee pains, depressive mood, exercise habits, social activity, social isolation, and frailty.Results　Awareness of the term "frailty" was estimated as 20.1% in total (15.5% in men and 24.3% in women). The subgroup with the highest "frailty" awareness was women who exercised, were socially active, and had a DVS of 4 or more (awareness of 36.3%). Significant independent correlates of "frailty" awareness were age (as per year: multivariate-adjusted prevalence ratio=1.03, [95% confidence interval=1.02-1.04]), sex (women: 1.35 [1.21-1.51]), educational attainment (high school: 1.27 [1.11-1.45], higher than junior college/vocational schools: 1.47 [1.28-1.70]), equivalent income (more than 2.5 million yen/year: 1.12 [1.01-1.25]), exercise habits (presence: 1.26 [1.11-1.43]), DVS (6 points or more: 1.37 [1.21-1.55]), social activity (presence: 1.33 [1.20-1.49]), social isolation (presence: 0.75 [0.67-0.85]), and frailty (presence: 0.72 [0.62-0.84]).Conclusions　Although many policies refer to "frailty," the level of awareness of the term among older adults was low. Older adults, especially women, who had higher socioeconomic status, better exercise and dietary habits, and stronger social connections, were significantly more aware of the term. In contrast, individuals who were socially isolated and/or frail did not know the term. Thus, it is crucial to develop specific measures to promote frailty prevention among high-risk individuals.

A Community-Wide Intervention Trial for Preventing and Reducing Frailty Among Older Adults Living in Metropolitan Areas: Design and Baseline Survey for a Study Integrating Participatory Action Research With a Cluster Trial.

BACKGROUND: Preventing and reducing frailty is an important challenge for Japan in the next decade, especially in metropolitan areas. We launched a community-wide intervention trial (the Ota Genki Senior Project) in 2016 to develop effective community-based strategies for frailty prevention in metropolitan areas. This report describes the study design and baseline survey. METHODS: This study is a community-wide intervention trial that integrates participatory action research into a cluster non-randomized controlled trial for adults aged 65 years or older living in Ota City, Tokyo. We allocated 3 of 18 districts to an intervention group and the other 15 to a control group. Using a mailed self-administered questionnaire, we conducted a baseline survey of 15,500 residents (8,000 and 7,500 in the intervention and control groups, respectively) from July through August 2016. In addition to socioeconomic status and lifestyle variables, we assessed frailty status (primary outcome) and physical, nutritional, and psychosocial variables (secondary outcomes). Based on the baseline findings, an intervention to improve outcomes will be implemented as participatory action research. Follow-up surveys will be conducted in the same manner as the baseline survey. RESULTS: A total of 11,925 questionnaires were returned (76.9% response rate; 6,105 [76.3%] and 5,820 [77.6%] in the intervention and control groups, respectively), and 11,701 were included in the analysis (mean age, 74.3 [standard deviation, 5.5] years; 48.5% were men). CONCLUSIONS: This study is expected to contribute to development of a prototype of a community-wide frailty prevention strategy, especially in metropolitan areas in Japan. TRIAL REGISTRATION: UMIN Clinical Trials Registry (UMIN000026515).

Individual- and community-level neighbor relationships and physical activity among older Japanese adults living in a metropolitan area: a cross-sectional multilevel analysis.

BACKGROUND: Informal neighbor relationships (NRs) are considered a structural aspect of social relationships. Although NRs might affect physical activity (PA), no previous study has simultaneously examined compositional and contextual associations of NRs with PA. In this study, we examined whether individual- and community-level NRs were independently associated with PA. METHODS: We analyzed cross-sectional data from 8592 (4340 men and 4252 women) non-disabled residents aged 65-84 years from all 18 districts of Ota City, Tokyo. PA was assessed by using the International Physical Activity Questionnaire-Short Form. In addition, we calculated moderate-to-vigorous PA (MVPA), its components (vigorous PA [VPA], moderate PA [MPA], and walking time [WT]), and sitting time (ST). Individual-level NRs were categorized as "visiting each other," "standing and chatting," "exchange of greetings," or "none." Community-level NRs were defined as the proportions of residents with active NRs (i.e., those in the categories visiting each other and standing and chatting) in the 18 districts. Using multilevel regression analyses, we examined independent associations of individual- and community-level NRs with PA variables and adjusted for important confounders. RESULTS: Individual-level NRs were consistently positively associated with MVPA and its components (VPA, MPA [in men], and WT) in both sexes, and the dose-response relationships were significant (all P < 0.041 for trend). In men, community-level NRs (by 1% estimation) were positively associated with individual MVPA (2.1 metabolic equivalent-hours/week, 95% confidence interval: 0.7-3.4), VPA (8.6 min/week, 2.7-14.4), and WT (11.6 min/week, 2.2-20.9), regardless of the degree of individual-level NRs. Significant cross-level interactions of NRs with MVPA and VPA were observed among men, and the dose-response relationships were significant (both P < 0.037 for trend). Neither individual- nor community-level NRs were associated with ST in either sex. CONCLUSIONS: Men and women with inaccessible neighbors engaged in less MVPA, while men living in communities with active NRs engaged in more MVPA, regardless of individual-level NRs. NRs at the individual and community level might help prevent physical inactivity among men.

Fourteen babies born after round spermatid injection into human oocytes.

During the human in vitro fertilization procedure in the assisted reproductive technology, intracytoplasmic sperm injection is routinely used to inject a spermatozoon or a less mature elongating spermatid into the oocyte. In some infertile men, round spermatids (haploid male germ cells that have completed meiosis) are the most mature cells visible during testicular biopsy. The microsurgical injection of a round spermatid into an oocyte as a substitute is commonly referred to as round spermatid injection (ROSI). Currently, human ROSI is considered a very inefficient procedure and of no clinical value. Herein, we report the birth and development of 14 children born to 12 women following ROSI of 734 oocytes previously activated by an electric current. The round spermatids came from men who had been diagnosed as not having spermatozoa or elongated spermatids by andrologists at other hospitals after a first Micro-TESE. A key to our success was our ability to identify round spermatids accurately before oocyte injection. As of today, all children born after ROSI in our clinic are without any unusual physical, mental, or epigenetic problems. Thus, for men whose germ cells are unable to develop beyond the round spermatid stage, ROSI can, as a last resort, enable them to have their own genetic offspring.

[Relationship between eating alone and dietary variety among urban older Japanese adults].

Objectives　This study aimed to investigate the practice of eating alone, including its frequency, among urban older Japanese adults and to clarify its association with dietary variety.Methods　In June 2016, self-reported questionnaires were mailed to 15,500 men and women aged 65 years or older living in Ota City, Tokyo without nursing care certification. Of these, 11,925 questionnaires were returned (a 76.9% response rate), and responses from 8,812 subjects were analyzed (a 56.9% valid response rate). The frequency of eating alone, defined as the number of days per week eating alone for each meal, was reported as zero, one to three, four to six, or seven days. A Dietary Variety Score (DVS) was obtained based upon consumption frequencies per week of 10 food groups. Total points were calculated, and three or fewer points indicated low dietary variety. The association between eating alone and low dietary variety was analyzed by binomial logistic regression, with adjustments for potential confounding variables including age, body mass index (BMI), location, educational history, equivalent income, employment, living alone, medical history, and smoking and drinking habits.Results　The results indicated that 47.1% of men and 48.5% of women ate alone at least one day per week, and 14.9% of men and 16.9% of women ate alone every day. For men, after adjusting for potential confounders (ORs=1.51-2.00), the odds ratios (ORs) of low DVSs were significantly higher among those who ate alone one to three days, four to six days, or seven days per week compared to those who ate with others. For women, adjusted ORs were not significant among those who ate alone every day (OR=1.15, 95% CI=0.92-1.43). Men and women who ate alone had significantly lower ORs of eating green or yellow vegetables, fruits, and fats/oils almost every day compared to those who ate with others.Conclusion　Among urban Japanese older adults, about 50% of subjects reported eating alone. Eating alone was associated with lower dietary variety scores regardless of age, income, and family status. Therefore, the present findings might be useful for developing strategies for addressing undernutrition among urban older adults eating alone.

[The impact of direct-acting antiviral therapy on the diagnosis of hepatitis-C virus-associated hepatocellular carcinoma].

Since the introduction of direct-acting antiviral (DAA)-based combination therapies in September 2014 for patients with chronic hepatitis-C (CH-C), numerous patients have been diagnosed with hepatitis-C virus (HCV)-associated hepatocellular carcinomas (HCCs) during the screening performed prior to DAA therapy. The present study was conducted on the antiviral therapy for CH-C in two phases:i) the interferon (IFN) phase between January 2011 and August 2014 and ii) the DAA phase between September 2014 and September 2016. During the DAA phase, HCCs were detected in eight patients who were referred to our hospital for anti-HCV therapy. In contrast, HCCs were detected in only two patients during the IFN phase. The number of patients with newly detected HCC in the DAA phase (20.5%) who were referred for the anti-HCV therapy was significantly higher than that in the IFN phase (1.7%). Owing to the high efficacy and safety of the DAA therapy, the number of patients referred to our hospital for anti-HCV therapy increased from 40.5 persons/year in the IFN phase to 80.3 persons/year in the DAA phase. The average ages of patients in the DAA and IFN phases were 68 and 61 years, respectively. The increase in the number of patients with newly detected HCC referred for the anti-HCV therapy in the DAA phase could be attributed to the increase in the number of referred patients for anti-HCV therapy and the aging of these patients in the DAA phase. All the eight patients with newly detected HCC who were referred for anti-HCV therapy in the DAA phase received curative treatments. The median age, rate of liver cirrhosis, and median tumor size of the patients were 69 years, 13%, and 16mm. Therefore, the findings of this study indicate that DAA therapies not only eradicate HCV infection but also contribute to the early diagnosis of HCC by encouraging the HCV-infected patients to visit hospitals and by promoting active network between hepatologists and family physicians.

Comparison of Absorbents and Drugs for Internal Decorporation of Radiocesium: Advances of Polyvinyl Alcohol Hydrogel Microsphere Preparations Containing Magnetite and Prussian Blue.

Radiocesium nuclides, used as a gamma ray source in various types of industrial equipments and found in nuclear waste, are strictly controlled to avoid their leakage into the environment. When large amounts of radiocesium are accidentally incorporated into the human body, decorporation therapy should be considered. Although standard decorporation methods have been studied since the 1960s and were established in the 1970s with the drug Radiogardase(®) (a Prussian blue preparation), application of recent advances in pharmacokinetics and ethical standards could improve these methods. Here we designed a modern dosage form of hydrogel containing cesium-absorbents to alleviate intestinal mucosa irritation due to the cesium-binding capacity of the absorbents. The effectiveness of the dosage form on fecal excretion was confirmed by quantitative mouse experiments. The total cesium excretion rate of the crystal form (1.37±0.09) was improved by the hydrogel form (1.52±0.10) at the same dose of Prussian blue, with a longer gastrointestinal tract transit time. Using a mouse model, we compared the effects of several drugs on fecal and urinary excretion of internal cesium, without the use of absorbents. Only phenylephrine hydrochloride significantly enhanced cesium excretion (excretion rate of 1.17±0.08) via the urinary pathway, whereas none of the diuretic drugs tested had this effect. These findings indicate that modifying the dosage form of cesium absorbents is important for the decorporation of internal radiocesium contamination.

Radiation increases the cellular uptake of exosomes through CD29/CD81 complex formation.

Exosomes mediate intercellular communication, and mesenchymal stem cells (MSC) or their secreted exosomes affect a number of pathophysiologic states. Clinical applications of MSC and exosomes are increasingly anticipated. Radiation therapy is the main therapeutic tool for a number of various conditions. The cellular uptake mechanisms of exosomes and the effects of radiation on exosome-cell interactions are crucial, but they are not well understood. Here we examined the basic mechanisms and effects of radiation on exosome uptake processes in MSC. Radiation increased the cellular uptake of exosomes. Radiation markedly enhanced the initial cellular attachment to exosomes and induced the colocalization of integrin CD29 and tetraspanin CD81 on the cell surface without affecting their expression levels. Exosomes dominantly bound to the CD29/CD81 complex. Knockdown of CD29 completely inhibited the radiation-induced uptake, and additional or single knockdown of CD81 inhibited basal uptake as well as the increase in radiation-induced uptake. We also examined possible exosome uptake processes affected by radiation. Radiation-induced changes did not involve dynamin2, reactive oxygen species, or their evoked p38 mitogen-activated protein kinase-dependent endocytic or pinocytic pathways. Radiation increased the cellular uptake of exosomes through CD29/CD81 complex formation. These findings provide essential basic insights for potential therapeutic applications of exosomes or MSC in combination with radiation.

Glycoprotein 2 as a gut gate keeper for mucosal equilibrium between inflammation and immunity.

Glycoprotein 2 (GP2) is a widely distributed protein in the digestive tract, contributing to mucosal barrier maintenance, immune homeostasis, and antigen-specific immune response, while also being linked to inflammatory bowel disease (IBD) pathogenesis. This review sheds light on the extensive distribution of GP2 within the gastrointestinal tract and its intricate interplay with the immune system. Furthermore, the significance of GP2 autoantibodies in diagnosing and categorizing IBD is underscored, alongside the promising therapeutic avenues for modulating GP2 to regulate immunity and maintain mucosal balance.

High-energy SR-XRF Imaging of Cesium and Trace Elements in Mouse Kidneys: Short Communication.

For understanding trace element dynamics in tissues, methods for analyzing elemental distribution and localization without destroying tissue structures and cell arrangements are desired. Synchrotron radiation X-ray fluorescence (SR-XRF) analysis is one of the non-destructive and multi-element simultaneous analyses. The kidney is the major excretion pathway of cesium (Cs) taken into the body, and an understanding of cesium distribution in the kidney would be useful for establishing technology to facilitate the excretion of radioactive Cs from the body due to nuclear disasters. In the present study, the distribution of cesium and trace elements, such as iron (Fe) and zinc (Zn), corresponding to the kidney structure was examined in Cs-administered mice by SR-XRF imaging with high-energy excitation X-rays (40 keV). By beam scanning with a 200-µm square beam, clear Cs images corresponding to the renal layer structure were obtained for the renal specimen at the early phase after Cs administration with the mean renal Cs concentration of 24.1 ± 3.2 µg/g. Cs was distributed mainly in the medulla and the outer stripe of the outer medulla located in the center area of the kidney. Unlike the Cs distribution, endogenous Fe and Zn tended to be lower in the medulla than in the outer stripe of the outer medulla and the cortex. This method is effective for analyzing Cs distribution because it can simultaneously analyze the distribution of endogenous trace elements.

[A case of HER2-positive and AFP-producing gastric cancer successfully treated by trastuzumab/docetaxel/S-1 combination therapy].

A 74-year-old man was referred to our hospital because of abdominal distension. Upper gastrointestinal endoscopy revealed advanced gastric cancer and early gastric cancer. HER2-positive and AFP-producing gastric cancer with peritonitis carcinomatosa showing no indication for operation was diagnosed by histopathological and radiological examinations. He was treated with trastuzumab, docetaxel, and S-1 combination chemotherapy. At the end of the second course of therapy, the primary lesion was remarkably decreased in size and was associated with a significant decrease in serum AFP level. No serious adverse events occurred except for grade 3-4 leukopenia and neutropenia. We carried out eight courses of chemotherapy. Trastuzumab, docetaxel, and S-1 combination chemotherapy promise to be one of the effective treatments for HER2-positive and AFP-producing gastric cancer that have no indication for radical cure excision.

Uranium chelating ability of decorporation agents in serum evaluated by X-ray absorption spectroscopy.

Internal exposure to actinides such as uranium and plutonium has been reduced using chelating agents for decorporation because of their potential to induce both radiological and chemical toxicities. This study measures uranium chemical forms in serum in the presence and absence of chelating agents based on X-ray absorption spectroscopy (XAS). The chelating agents used were 1-hydroxyethane 1,1-bisphosphonate (EHBP), inositol hexaphosphate (IP6), deferoxamine B (DFO), and diethylenetriaminepentaacetate (DTPA). Percentages of uranium-chelating agents and uranium-bioligands (bioligands: inorganic and organic ligands coordinating with uranium) dissolving in the serum were successfully evaluated based on principal component analysis of XAS spectra. The main ligands forming complexes with uranium in the serum were estimated as follows: IP6 > EHBP > bioligands > DFO ≫ DTPA when the concentration ratio of the chelating agent to uranium was 10. Measurements of uranium chemical forms and their concentrations in the serum would be useful for the appropriate treatment using chelating agents for the decorporation of uranium.

An investigation of the potential effect of vacuoles in human sperm on DNA damage using a chromosome assay and the TUNEL assay.

BACKGROUND: The aims of this study were to establish whether individual differences exist in the frequency and size of vacuoles found in human sperm and to ascertain whether such vacuoles are involved in causing DNA damage. METHODS: Morphologically normal sperm were obtained from 15 IVF and 2 ICSI patients and 3 fertile donors. (i) Sperm heads were analyzed for the presence of vacuoles under a 1000× differential interference contrast microscope. (ii) In three patients and two donor samples, structural chromosomal damage was evaluated in normal sperm containing large vacuoles and selected at 1000× magnification for injection into mouse oocytes. (iii) In 10 patients and two donor samples, confocal laser microscopy detected DNA damage in sperm-exhibiting large vacuoles and stained with an in situ cell death detection kit. RESULTS: (i) Vacuoles were observed in almost all normal sperm from patient and donor ejaculates and were mainly located at the tip or middle area of the sperm heads. However, average incidence of normal sperm exhibiting large vacuoles was 4.6 and 4.2% in the patient and donor groups, respectively. (ii) Sperm chromosome assays did not reveal any differences in the incidence of structural chromosome aberrations between sperm exhibiting large vacuoles and those without them (9.1 versus 4.1%). (iii) No significant difference in frequency of TUNEL-positive cells was found between normal sperm with large vacuoles and those without them in the samples examined. Among 227 sperm exhibiting large vacuoles, only 7 cells were TUNEL positive. CONCLUSION: The results showed that large vacuoles were not responsible for DNA damage, suggesting that intra-cytoplasmic injection of morphologically selected sperm may not be required for patients who produce high-quality semen.

Patterned growth of graphene over epitaxial catalyst.

Rectangle- and triangle-shaped microscale graphene films are grown on epitaxial Co films deposited on single-crystal MgO substrates with (001) and (111) planes, respectively. A thin film of Co or Ni metal is epitaxially deposited on a MgO substrate by sputtering while heating the substrate. Thermal decomposition of polystyrene over this epitaxial metal film in vacuum gives rectangular or triangular pit structures whose orientation and shape are strongly dependent on the crystallographic orientation of the MgO substrate. Raman mapping measurements indicate preferential formation of few-layer graphene films inside these pits. The rectangular graphene films are transferred onto a SiO(2)/Si substrate while maintaining the original shape and field-effect transistors are fabricated using the transferred films. These findings on the formation of rectangular/triangular graphene give new insights on the formation mechanism of graphene and can be applied for more advanced/controlled graphene growth.

Microarray-based global mapping of integration sites for the retrotransposon, intracisternal A-particle, in the mouse genome.

Mammalian genomes contain numerous evolutionary harbored mobile elements, a part of which are still active and may cause genomic instability. Their movement and positional diversity occasionally result in phenotypic changes and variation by causing altered expression or disruption of neighboring host genes. Here, we describe a novel microarray-based method by which dispersed genomic locations of a type of retrotransposon in a mammalian genome can be identified. Using this method, we mapped the DNA elements for a mouse retrotransposon, intracisternal A-particle (IAP), within genomes of C3H/He and C57BL/6J inbred mouse strains; consequently we detected hundreds of probable IAP cDNA-integrated genomic regions, in which a considerable number of strain-specific putative insertions were included. In addition, by comparing genomic DNAs from radiation-induced myeloid leukemia cells and its reference normal tissue, we detected three genomic regions around which an IAP element was integrated. These results demonstrate the first successful genome-wide mapping of a retrotransposon type in a mammalian genome.

Metaphase II karyoplast transfer from human in-vitro matured oocytes to enucleated mature oocytes.

Metaphase II karyoplast transfer is believed to be a useful method to rescue aged oocytes. This study attempted karyoplast transfer of in-vitro matured metaphase II (MII) oocytes, as a model of aged oocytes, into enucleated freshly ovulated metaphase II oocytes with visualization of their chromosomes under an inverted microscope. Recipient karyoplasts derived from immature oocytes were cultured in-vitro until first polar body extrusion. After 1-2 days culture, 52.1% extruded a polar body, 95.5% had PSC, aneuploidy was very low (4.5%) and none had structural aberrations. Donor oocytes were obtained from IVF or intracytoplasmic sperm injection (ICSI) patients. Chromosomes were easily confirmed in 92.3% and 95.0% of in-vivo and in-vitro matured oocytes respectively. Thirty-one karyoplasts were placed in the perivitelline space of enucleated donor oocytes, and 25 (80.6%) fused to form a reconstituted oocyte. Fertilization, cleavage and blastocyst formation rates following ICSI were 76.0%, 64.0% and 28.0% respectively for reconstructed oocytes and 59.2%, 48.0% and 3.1% respectively for control (in-vitro matured) oocytes. Chromosomal analysis of five embryos developed after karyoplast transfer and ICSI showed normal diploid sets of 46 chromosomes. In conclusion, this metaphase II karyoplast transfer technique can be applied to the solution of chromosomal abnormalities related to oocyte ageing.