Normal calves produced after transfer of embryos cultured in a chemically defined medium supplemented with epidermal growth factor and insulin-like growth factor I following ovum pick up and in vitro fertilization in Japanese black cows.

The objective of this study was to examine whether high concentrations of epidermal growth factor (EGF) and/or insulin-like growth factor I (IGF-I) would have a beneficial effect on bovine embryo development in vitro and to obtain normal calves by using an ovum pick up method and embryo culture in a chemically defined medium. When compared with controls, EGF (100 or 200 ng/ml) or IGF-I (50 or 100 ng/ml) significantly increased the rate of embryos that developed into blastocysts during an 8-day culture after the in vitro fertilization of oocytes obtained from ovaries from a slaughterhouse. IGF-I induced a dose-dependent increase in cell number in both the inner cell mass and the trophectoderm, whereas EGF stimulated proliferation only in the inner cell mass. A combination of EGF (100 ng/ml) and IGF-I (50 ng/ml) produced an additive effect, and embryos developed into blastocysts at a comparatively high rate (27.9%) compared with controls (12.0%). A similar rate of development was achieved using a combination of EGF and IGF-I in the culture of embryos following ovum pick up by ultrasound-guided transvaginal follicular aspiration and in vitro fertilization, and 5 blastocysts that developed after the culture were transferred into uteri; two embryos implanted, and normal calves were born. These results suggest that the combined use of EGF and IGF-I makes bovine embryo culture in a chemically defined medium a practical and useful procedure for producing blastocysts, and its application to embryo culture following ovum pick up and in vitro fertilization could be useful for producing normal calves.

Evaluating Methods for Detecting Escherichia albertii in Chicken Meat.

ABSTRACT: Escherichia albertii is an emerging foodborne pathogen. The source of the E. albertii infection in most foodborne outbreaks is unknown because E. albertii is difficult to isolate from suspected food or water. E. albertii has a broad host range among birds and can be isolated from chicken meat. In this study, PCR assay, enrichment, and isolation conditions for detecting E. albertii in chicken meat were evaluated. The growth of 47 E. albertii strains isolated in Japan between 1994 and 2018 and a type strain was evaluated in modified EC broth (mEC) and mEC supplemented with novobiocin (NmEC) and on media containing carbohydrates. The enzyme used for the nested PCR, the enrichment conditions, the most-probable-number (MPN) method, and agar media were also evaluated with chicken meat. To distinguish E. albertii from presumptive non-E. albertii bacteria, desoxycholate hydrogen sulfide lactose agar (DHL), MacConkey agar (MAC), and these agars supplemented with rhamnose and xylose (RX-DHL and RX-MAC, respectively) were used. All E. albertii strains grew in mEC and NmEC at both 36 and 42°C and did not utilize rhamnose, sucrose, or xylose. Both the first and nested PCRs with TaKaRa Ex Taq, which was 10 to 100 times more active than the other enzymes, produced positive results in enrichment culture of 25 g of chicken meat inoculated with >20 CFU of E. albertii and incubated in mEC and NmEC at 42°C for 22 ± 2 h. Thus, the first PCR was sensitive enough to detect E. albertii in chicken meat. The MPN values in mEC and NmEC were 0.5- and 2.3-fold higher than the original inoculated bacterial levels, respectively. E. albertii in chicken meat was more efficiently isolated with enrichment in NmEC (70.1 to 100%) and plating onto RX-DHL (85.4%) and RX-MAC (100%) compared with enrichment in mEC (53.5 to 83.3%) and plating onto DHL (70.1%) and MAC (92.4%). Thus, optimized conditions for the surveillance of E. albertii contamination in food and investigations of E. albertii outbreaks, including the infectious dose, were clarified.

Obstructive sleep apnea-hypopnea syndrome patients with overweight and hypertension in a Japanese workplace.

The objective of this study was to determine the relationship between obstructive sleep apnea-hypopnea syndrome (OSAHS) and overweight combined with hypertension and to examine whether OSAHS in conjunction with overweight and hypertension is associated with daytime sleepiness. In a Japanese workplace of 28,636 employees, 368 men (19-62 yr old), who were anxious regarding their OSAHS symptoms, underwent home pulse oximetry. Of these, 153 men subsequently underwent all-night polysomnography (PSG), and OSAHS was diagnosed in 149. We next classified these 149 men into the following groups: A [Overweight (-)/Hypertension (-), n=41], B [Overweight (-)/Hypertension (+), n=15], C [Overweight (+)/Hypertension (-), n=46], and D [Overweight (+)/Hypertension (+), n=47]. The Epworth Sleepiness Scale (ESS) was used to evaluate daytime sleepiness and the apnea-hypopnea index (AHI) was used to evaluate the severity of OSAHS. The averages of the ESS score and the AHI were compared in each group. Both the average ESS scores and the percentage of ESS scores > or =11 were not significantly different among the groups. The average AHI of group D was the highest among all of the groups and that of group C was significantly higher than those of groups A and B. In all the groups, the OSAHS patients with overweight and hypertension in this study had the highest AHI. The level of daytime sleepiness evaluated by the ESS in this study was almost the same in the OSAHS patients regardless of the degree of overweight or hypertension. These observations suggest that it is necessary to positively recommend PSG to men who are suspected of having OSAHS with overweight and hypertension, even if they do not have daytime sleepiness.

Ventricular Rhythm and Hypotension in a Patient with Pheochromocytoma-induced Myocardial Damage and Reverse Takotsubo Cardiomyopathy.

A 33-year-old woman experienced near-syncope at a hospital. Electrocardiography revealed an intermittent ventricular rhythm. The echocardiogram at admission indicated mild hypokinesis and severe diffuse hypokinesis with reverse takotsubo cardiomyopathy on the following day. The patient experienced abdominal pain on the admission day, and computed tomography identified a large left adrenal mass. Her catecholamine levels increased remarkably on the third day. The wall motion improved on the twelfth day. The tumor was successfully resected and the patient was diagnosed with an ectopic pheochromocytoma. The ventricular rhythm with myocardial damage and hypotension induced by the reverse takotsubo cardiomyopathy masked the characteristic symptoms of pheochromocytoma.

Improvement of preimplantation development of in vitro-fertilized bovine zygotes by glucose supplementation to a chemically defined medium.

The influences of glucose supplementation on early development of bovine embryos in BSA-free synthetic oviduct fluid were examined. Among the groups supplemented with 1.5, 2.0, 4.0 or 5.6 mM glucose either at 0, 72 or 144 hr after fertilization, blastocysts yield significantly increased in the group supplemented with 4.0 mM glucose 144 hr after fertilization compared to the controls without glucose supplementation. The results suggest that appropriate amounts of glucose supplemented to the medium at the specific stage of embryo culture may be useful for the production of bovine blastocysts.

Recurrence of ventricular fibrillation in a patient with non-type 1 Brugada electrocardiographic morphology.

A healthy 25-year-old man suffered from loss of consciousness due to ventricular fibrillation (VF). Emergency services required multiple cardioversion to restore sinus rhythm. Repeated electrocardiographic (ECG) recordings after admission showed non-type 1 Brugada ST-segment elevation in V1 and V2. Intravenous pilsicainide infusion augmented the ST-segment elevation but its morphology did not represent type-1 ECG. Intravenous administration of isoproterenol normalized the ST-segment elevation, and programmed electrical stimulation induced VF. Spontaneous VF recurred 1 year after introduction of implantable cardioverter defibrillator. Non-type 1 ST-segment elevation, to which pharmacological responses are similar to Brugada syndrome, may be used as a hallmark of ventricular tachyarrhythmia.

Syntheses and receptor-binding studies of derivatives of the opioid antagonist naltrexone.

Naltrexone (1), which is a member of the group of competitive opioid antagonists, shows a strong affinity for mu-receptors and its derivatives have been notable as novel receptor antagonists. In this paper, the preparation of several naltrexone derivatives is described; these were used to investigate the role of the oxygenated functional groups in facilitating binding to a series of the opioid receptors. The derivatives showed affinity for opioid mu-receptors which was similar to that of naltrexone, but these compounds, which had masked hydroxyl functional groups, displayed a moderate activity. These results suggest that every oxygenated functional group in naltrexone (1) plays an important role in binding to the opioid receptor.