Comparing pig breeds with genetically low and high backfat thickness: differences in expression of adiponectin, its receptor, and blood metabolites.

Here we characterized gene expressions in subcutaneous adipose tissue and blood metabolites of pigs with genetically low backfat (Landrace) and high backfat (Meishan). As pigs aged from 1 wk-to 3-mo old, mRNA levels of adipose-specific genes increased, although their gene expressions coding for major enzymes involved in lipid metabolism (lipoprotein lipase, fatty acid synthase, and hormone-sensitive lipase) did not differ between lean and fat pigs. Instead, there were significant effects for adiponectin and its receptor AdipoR1 mRNA levels between the two breeds of which respective expressions were lower and higher in Meishan by 3 mo of age. Contrary to changes in gene expressions, the concentrations of blood glucose, triglyceride (TG), and NEFA in both breeds decreased during growth, and 3-mo-old Meishan evidenced lower glucose with higher TG than the Landrace. The homeostasis model assessment insulin resistance (HOMA-IR) index was also calculated from the measurements of fasting glucose and insulin concentration, and Meishan showed a higher value than the Landrace. We next examined these differences in Landrace and Meishan crossbreds, which were phenotypically distinguishable by the backfat thickness as the former lean type and the latter fat type. As with the purebreds, high backfat Meishan crosses showed the characteristics of lower glucose and higher TG in circulating levels and also lower adiponectin transcripts in subcutaneous adipose tissue. Collectively, our results demonstrate that levels of adiponectin and its receptor gene expressions, blood glucose, blood lipids, and HOMA-IR in pigs vary between lean and fat. These observations strongly suggest the possibility that overall metabolic differences rather than adipocyte ability itself contribute to the fatness of genetically high backfat pigs.

Thiamine accumulation and thiamine triphosphate decline occur in parallel with ATP exhaustion during postmortem aging of pork muscles.

We aimed to clarify the mechanisms affecting postmortem thiamine and its phosphoester contents in major edible pork muscles, namely the longissimus lumborum (LL) in addition to vastus intermedius (VI). Metabolomic analysis by capillary electrophoresis-time of flight mass spectrometry revealed that the level of thiamine triphosphate (ThTP), approximately 1.8-fold higher in LL than in VI muscle at 0h postmortem, declined in the first 24hrs, resulting in an undetectable level at 168h postmortem in both muscles. In contrast, the thiamine content in both muscles increased after 24h postmortem during the aging process. The thiamine accumulation and ThTP decline progressed in parallel with a drastic reduction of the ATP level. The intermuscular differences in pH at 24h and in expression of thiamine transporter and thiamine pyrophosphokinase might result in delayed thiamine generation in LL. These results suggest that postmortem ATP exhaustion forced ThTP hydrolysis and further depyrophosphorylation of thiamine diphosphate in the porcine muscles, which resulted in thiamine accumulation.

Use of smartphone attached mobile thermography assessing subclinical inflammation: a pilot study.

OBJECTIVE: To verify the reliability and validity of FLIR ONE, a device connected to a smartphone, for the assessment of inflammation based on relative temperature increase compared with the thermography routinely used in pressure ulcer (PU) and diabetic foot assessment. METHOD: Participants in this pilot cross-sectional observational study were recruited from the patients in the PU team rounds and the diabetic foot outpatient clinic at the university hospital in January 2015. Cohen's kappa coefficient with its 95% confidence intervals was used to evaluate the criterion-related validity and inter- and intra-rater reliability for the thermal imaging assessment. For assessing criterion-related validity, a hand-held high-end infrared thermography device was used to provide reference data. Comparison of thermal images between the smartphone-connected device and the hand-held device was performed with both a 'predetermined range' and an 'automatically-set range.' For assessing inter-rater reliability, two assessors evaluated the thermal images taken by the mobile thermography. For assessing intra-rater reliability, one assessor evaluated the thermal images twice. The thermal images were shown to the assessors at random. RESULTS: Among 16 thermal images obtained from eight patients, kappa coefficients for each value were as follows: for the predetermined range and automatically-set range, respectively, the criterion-related validity was 1.00 (95% confidence interval 1.00-1.00) and 1.00 (95% confidence interval 1.00-1.00); the inter-rater reliability was 1.00 (95% confidence interval 1.00-1.00) and 1.00 (95% confidence interval 1.00-1.00); and the intra-rater reliability was 1.00 (95% confidence interval 1.00-1.00) and 1.00 (95% confidence interval 1.00-1.00). CONCLUSION: This pilot study suggests that FLIR ONE can work as an alternative device for assessing subclinical inflammation in PUs and the diabetic foot in clinical settings. Our results may facilitate clinicians in accepting the routine use of thermal imaging assessment at the patients' bedside.

Distribution of tropomyosin isoforms in different types of single fibers isolated from bovine skeletal muscles.

To clarify the relationship between myosin heavy chain (MyHC) isoforms and tropomyosin (TPM) isoforms in single fibers, 64 single fibers were isolated from each of bovine three muscles (masseter, semispinalis and semitendinosus). mRNA expressions of MyHC and TPM isoforms were analyzed by real-time PCR. All single fibers from the masseter expressed MyHC-slow. The fibers from the semispinalis expressed both MyHC-slow and 2a. The fibers from the semitendinosus expressed MyHC-slow, 2a and 2x. TPM-1 and TPM-2 were co-expressed in 2a and 2x type fibers, and TPM-2 and TPM-3 were co-expressed in slow type fibers. The expression pattern of TPM isoforms in each fiber type was similar between fibers isolated from different muscles. These results suggest that TPM-1 and TPM-3 isoforms correspond to the function of 2a or 2x type fibers and slow type fibers, respectively, with TPM-2 in common. Furthermore, the patterns of MyHC and TPM isoform combinations did not vary among single fibers isolated from the individual muscles examined.

Profiling of differentially expressed microRNA and the bioinformatic target gene analyses in bovine fast- and slow-type muscles by massively parallel sequencing.

MicroRNA (miRNA) are highly conserved, noncoding small RNA involved in post-transcriptional gene regulation in a variety of biological processes. To elucidate roles of miRNA in bovine muscle type specification and maintenance, we sought to determine differentially expressed miRNA between semitendinosus (STD) and masseter (MS) muscles from 3 Japanese black cattle by massively parallel sequencing. Differential gene expression of myosin heavy chain (MyHC) isoforms confirmed that STD and MS were MyHC-2x- and MyHC-1-abundant muscles, respectively. In total, 192 known miRNA and 20 potential new bovine miRNA were obtained from the sequencing. The differentially expressed miRNA with more than 2-fold difference in each muscle were identified. In particular, miR-196a and miR-885 were exclusively expressed in STD muscle, which was validated by quantitative reverse transcription-PCR (P=0.045 and P<0.001, respectively), whereas a slow type-directing miR-208b was highly expressed in MS compared with STD (false discovery rate<0.05). In addition, 16 potential novel miRNA were mapped and confirmed for their precursor structures by computational analyses. The results of functional annotation combined with in silico target analysis showed that the predicted target genes of miR-196a/b and miR-885 enriched gene ontology (GO) terms related to skeletal system development and regulation of transcription, respectively. Moreover, GO terms enriched from predicted targets miRNA suggested that STD-abundant- and MS-abundant-miRNA were associated with embryonic body planning and organ/tissue pattern formation, respectively. The present results revealed that the differentially expressed miRNA between the STD and MS muscles may play key roles to determine muscle type-specific tissue formation and maintenance in cattle thorough attenuating putative target genes involved in different developmental events.

Thermographic findings in a case of type 2 diabetes with foot ulcer and osteomyelitis.

Using thermography, skin temperature was evaluated in a 76-year-old patient with type II diabetes mellitus, presenting with diabetic foot ulceration on the right hallux and a corn on the left fourth toe. Increased skin temperature was observed in both the right hallux and the left fourth toe, though there were no visible clinical signs of infection. Unexpectedly, the high temperature area was seen to extend from the left fourth toe to the ankle. The patient was later diagnosed with osteomyelitis, due to the presence of a high-intensity area on T2-weighted magnetic resonance imaging, suggesting the elevated skin temperature was due to osteomyelitis. Based on these observations, thermography could prove useful for screening for foot ulcers with osteomyelitis.

Differential expression of the skeletal muscle proteome in grazed cattle.

The objective of this study was to investigate the differences in the muscle proteome of grass-fed and grain-fed cattle. Eight Japanese Black Cattle 10 mo of age were separated randomly into 2 groups: 1) grazing (grass-fed) and 2) concentrate (grain-fed) groups. All cattle were first housed individually in a stall barn and fed a combination of concentrate ad libitum and Italian ryegrass hay until 21 mo of age. After this control period, the 4 grass-fed cattle were placed on outdoor pasture, whereas the other 4 grain-fed cattle continued on the concentrate diet. The cattle were slaughtered at 27 mo of age, and tissues from the semitendinosus muscle were obtained for use in proteome analysis. Differential expression of muscle proteins in the 2 groups was carried out using 2-dimensional gel electrophoresis (2DE) and Western blot analyses, with subsequent mass spectrometry. Approximately 200 individual protein spots were detected and compared in each group using 2DE, of which 20 and 9 spots, respectively, showed differences in the spot intensity for the sarcoplasmic fraction and myofibrillar fraction. In the grazing group, the relative intensity of spots was significantly greater for adenylate kinase 1 and myoglobin in the sarcoplasmic fraction, and for slow-twitch myosin light chain 2 in the myofibrillar fraction (P < 0.05), than the concentrate group. The relative spot intensity of several glycolytic enzymes was significantly greater in the grazing group, such as beta-enolase 3, fructose-1,6-bisphosphate aldolase A, triosephosphate isomerase, and heat shock 27 kDa protein (P < 0.05). Moreover, significantly greater slow twitch of troponin T, troponin I, and myosin heavy chain of semitendinosus muscle was detected in the grazing group than in the concentrate group using Western blot analysis (P < 0.05). Several previous reports have described that the slow-twitch muscle contents affect elements of nutrition, flavor, and food texture of meat. This study revealed muscle fiber type conversion to slow-twitch tissues from fast-twitch tissues occurring with change in the energy metabolic enzyme when cattle were grazed in the latter fattening period. Although analyses of the influence on elements of nutrition, flavor, and food texture were not done for this study, these results show that slow-twitch converted muscle resulting from the grazing of cattle might modify several meat characteristics.

Muscle type specific expression of tropomyosin isoforms in bovine skeletal muscles.

Nucleotide sequences encoding an entire coding region for bovine tropomyosin (TPM) isoforms were determined. Three TPM isoforms, TPM1, TPM2 and TPM3, were expressed in bovine skeletal muscles, and exhibited a 93.3%, 99.6% and 100% amino acid homology to the human sequence, respectively. Based on the sequences, the composition of TPM isoforms was analyzed on cDNA and protein levels from five physiologically different muscles (masseter, diaphragm, psoas major, longissimus thoracis and semitendinosus) using RT-PCR and proteome analyses. Although the content of TPM2 was constantly about 50% of the total TPM in all muscles, the contents of TPM1 and TPM3 were different in muscles according to their function in muscle contraction. In masseter, the content of TPM3 cDNA was about 50% and higher than that of other muscles. In longissimus thoracis and semitendinosus, the contents of TPM1 cDNA were 29.6% and 31.7%, respectively, which were comparatively higher than that of other muscles. The result suggests that the TPM dimer consists of the TPM2 subunit regularly and TPM1 or TPM3 depending on whether the muscle is fast or slow type, respectively.

Intestinal ischemia induces late preconditioning against myocardial infarction: a role for inducible nitric oxide synthase.

OBJECTIVE: We tested the hypothesis that occlusion of the superior mesenteric artery induces late preconditioning against myocardial infarction and examined the effects of pharmacological modifiers of inducible nitric oxide synthase activity on the late preconditioning in anesthetized rats. METHODS: Rats underwent an intestinal ischemia preconditioning protocol (30 min occlusion of the superior mesenteric artery) or were sham-operated. They were subjected to a sustained 30 min of coronary occlusion and 180 min of reperfusion 24 h later. RESULTS: In rats receiving no pharmacological intervention, the percentage of myocardial infarct within the area at risk and left ventricle was 72+/-4% and 31+/-2%, respectively, in sham-operated rats, and these were significantly reduced to 44+/-4% and 23+/-2% (P<0.01) 24 h after intestinal ischemia preconditioning. Myeloperoxidase activity was significantly reduced by intestinal ischemia preconditioning. Administration of aminoguanidine (300 mg/kg, s.c.) or S-methylisothiourea sulfate (3 mg/kg, i.v.), both relative inducible NO synthase inhibitors, 60 or 30 min before sustained myocardial ischemia not only abolished the late preconditioning afforded by intestinal ischemia, but also inhibited the ability of intestinal ischemia preconditioning to significantly reduce neutrophil infiltration. A change in inducible NO synthase activity was not observed in normal myocardium 24 h after intestinal ischemia, but 30 min of coronary occlusion significantly increased the inducible NO synthase activity in the preconditioned group, which was abolished by aminoguanidine or S-methylisothiourea sulfate. CONCLUSIONS: These data provide pharmacological evidence that induction of inducible nitric oxide synthase, following intestinal ischemia, is associated with increased myocardial tolerance to infarction 24 h later.

Effects of prostaglandins on baroreflex during reperfusion of the ischaemic myocardium.

1. The present study was planned to: (i) determine whether the baroreflex control of heart rate (HR) and renal sympathetic nerve activity (RSNA) was attenuated during reperfusion of short-term ischaemic myocardium; and (ii) study whether blockade of prostaglandin synthesis with indomethacin reversed the inhibitory baroreflex. 2. Arterial pressure was lowered with intravenous sodium nitroprusside before coronary occlusion and 3 min after release of a 5 min occlusion of the left circumflex coronary artery in anaesthetized rabbits. The protocol was repeated 20 min after indomethacin (5 mg/kg, i.v.) or indomethacin vehicle (50 mmol/L tris(hydroxymethyl)aminomethane buffer, pH 8.4) treatment. In addition, this study was performed in a group of vagotomized rabbits. 3. Before indomethacin treatment, the slope of the mean arterial pressure (MAP)-RSNA relationship decreased from -3.3+/-0.77 to -2.01+/-0.69% change in RSNA/mmHg (P < 0.05) during reperfusion of ischaemic myocardium in intact rabbits. The decrease in the slope was reversed by administration of indomethacin. However, the decrease in the slope was not reversed by indomethacin vehicle. Furthermore, the reduction in the slope of the MAP-RSNA relationship during reperfusion of ischaemic myocardium was abolished in vagotomized rabbits. However, there was no inhibition of the slope of the MAP-HR relationship during reperfusion of ischaemic myocardium in either intact or vagotomized rabbits. 4. In conclusion, our data suggest that prostaglandins released by ischaemic myocardium can attenuate the baroreflex-mediated response of RSNA to lowered arterial pressure via vagal afferents during reperfusion of short-term ischaemic myocardium.

Serial MRI of cerebral infarcts before and after removal of an atrial myxoma.

We describe a cerebral infarct caused by atrial myxoma. A 30-year-old woman with an atrial myxoma presented with a right hemisensory deficit. MRI carried out before and after removal of the myxoma, showed multiple small bilateral white-matter infarcts which increased in number up to the surgery. A lesion in the left thalamus, which enlarged and showed contrast enhancement 4 months before surgery, resembled an old cerebral infarct by the time the myxoma was removed. The number of lesions stopped increasing after surgery. We suggest that atrial myxomas should be removed even in asymptomatic patients, to prevent cerebral infarcts due to embolism of tumour or thrombus.

Video-assisted thoracoscopic pericardiectomy for severe pericardial effusions.

Video-assisted thoracic surgery (VATS) has proven to be extremely useful in the diagnosis and treatment of multiple thoracic problems. We used the VATS technique to successfully treat refractory pericardial effusions associated with pericarditis by performing pericardiectomy.

Presumption of a suspect based on the skin fragment attached the rope.

An eight-year-old girl (second-year of elementary school) was strangled in an irrigation canal near her house and was left unconscious. The body of a male who hanged himself to death was discovered on the next day. The hanging mark on the neck of the man and the ligature mark on the neck of the girl were caused by double nooses of a rope, the width of the rope mark was nearly the same, and similar Zwischenkammblutung (subcutaneous bleeding in the middle of the rope mark) was observed in both cases. Therefore, we evaluated whether the marks were caused by the same rope. As a result, the DNA type of the skin fragment (keratinized epithelial cells) attached to the rope used by the man to hang himself was identical to that of the girl. On the basis of this evidence, the man who hanged himself to death was presumed to be the suspect who strangled the girl.

Community-based care for the mentally disordered in Kawasaki City: experiences from Kawasaki City Psychiatric Rehabilitation Center.

The basic aims of Kawasaki City Psychiatric Rehabilitation Center are: (i) taking some measures for the hospitalized patients unable to leave because of residential problems; (ii) providing a daily program for the discharged from psychiatric hospital; (iii) developing vocational rehabilitation for the mentally disordered; and (iv) developing a support network for the mentally disordered in the community. To fulfill these aims and to meet the various needs of our users, we have provided a very comprehensive program composed of a ward, outpatient section, day care groups, vocational service and half-way house. According to the program, various services such as medical treatment in a narrow sense, basic training of social skills and human relations, vocational training system that meets the various needs and work abilities of the users, and after care services that enable them to support themselves living in the community are available. These services give them great comfort to live in the community; however, they might serve to increase their dependency on the center. Our future task is to cooperate with various social resources such as workshops, group homes and places to stay and rest during the day, which are being developed in Kawasaki City. By supporting those facilities, we expect to refer our members to them and we put more weight on the indirect service than the direct one, which would hopefully enable us to cover and care for more people in need.

Genomic diversity in the pfoA region of the theta-toxin-deficient strains of Clostridium perfringens.

The genomic structure of the pfoA-colA region in six theta-toxin-deficient strains of Clostridium perfringens was examined by Southern hybridization using the pfoR, pfoA, pbg, arcABDC and colA genes, encoding regulator for pfoA, theta-toxin, beta-galactosidase, arginine metabolism enzymes and kappa-toxin, respectively, as gene probes. It is suggested that the productivity of theta-toxin in these strains is diverse because of the multiple genetic backgrounds including single deletion of pfoA, large deletion of the pfoA-colA region and the putative point mutations.

Collagenase gene (colA) is located in the 3'-flanking region of the perfringolysin O (pfoA) locus in Clostridium perfringens.

The 3'-flanking region of the beta-galactosidase gene (pbg), which is located downstream of the perfringolysin O gene (pfoA), and the 5'-flanking region of the collagenase gene (colA) of Clostridium perfringens strains NCTC8237 and 13, respectively, were analyzed. Southern analysis revealed that the colA gene is located 6.5 kb downstream of the pbg gene in the chromosome of C. perfringens. Sequence analysis showed that between the pbg and colA genes were the arcABDC and ahrC genes, whose putative products were quite similar to enzymes of the arginine deiminase pathway of Pseudomonas aeruginosa and the arginine repressor/activator of Bacillus subtilis, respectively. It is concluded that the genomic structure of the pfoA-colA region consists of pfoR-pfoA-ORF54-pbg-arcABDC-ahrC-colA.

[General outcomes of the fourth migration survey].

PIP: Results are presented from the fourth national migration survey carried out in Japan in 1996. The survey, which involved 40,400 individuals, covered a national sample of 14,083 households. The survey "was designed to gather information on various features of respondents' migration, such as time of in-migration to the present address, reasons for in-migration, location of the previous residence, life-time migration experiences, location of residence 1 and 5 years ago, and prospects of future migration." (EXCERPT)^ieng

Protamine-induced hypotension in heart operations: application of the concept of ventricular-arterial coupling.

Protamine sulfate often causes hypotension during heparin neutralization. The concept of ventricular-arterial coupling was applied to determine whether a negative inotropic effect or a vasodilating effect of protamine was the major contributing factor to this hypotension. Thirty-five patients who underwent cardiac operations were studied during operation by measuring instantaneous left ventricular pressure and aortic flow to examine the end-systolic pressure-volume relationship. We obtained end-systolic elastance and effective arterial elastance values in a beat-to-beat fashion with a single-beat estimation method. In 28 of the 35 patients (80%), mean arterial pressure decreased more than 10 mm Hg with protamine infusion. Parameters were compared at the following three points: before a decrease in mean arterial pressure (control), at maximally decreased mean arterial pressure (maximum), and at a middle point between control and maximum values (midpoint). At both midpoint and maximum, mean arterial pressure decreased significantly (control 79.6 +/- 12.6 mm Hg, midpoint 66.5 +/- 10.8 mm Hg, maximum 52.7 +/- 9.9 mm Hg; p < 0.01). Similar changes were observed in effective arterial elastance (control 2.00 +/- 0.75 mm Hg/ml, midpoint 1.60 +/- 0.53 mm Hg/ml, maximum 1.31 +/- 0.46 mm Hg/ml; p < 0.01). Although the decrease in end-systolic elastance at midpoint (control 3.08 +/- 1.61 mm Hg/ml, midpoint 2.92 +/- 1.68 mm Hg/ml) did not reach statistical significance, end-systolic elastance significantly decreased at maximum (2.63 +/- 1.46 mm Hg/ml; p < 0.01). Continuous measurements showed that the decreases in mean arterial pressure and effective arterial elastance always preceded the depression of end-systolic elastance and that afterload reduction by vasodilating effect of protamine was the mechanism most likely to have initiated the hypotension. Delayed decrease in contractility may be ascribed to reduced coronary perfusion pressure caused by vasodilation or to a direct effect of protamine.

[Household changes in Japan: major findings of the Third National Survey, 1994].

PIP: The results from the Third National Survey on Household Changes carried out in Japan in 1994 are presented. The survey involved a nationally representative sample of 8,578 households including 20,788 individuals. The results confirm a general trend toward the nuclear family, a reduction in average family size associated with demographic aging, a decline in sibship size associated with the fertility decline, and a rise in the age of leaving the parental home. (SUMMARY IN ENG)^ieng

Demonstrations of de-and remineralization mechanism as revealed in synthetic auditory ossicle (Apaceram) of rats by laser-Raman spectrometry.

In investigations of the process of bone destruction caused by chronic otitis media complicated with cholesteatoma, we proposed previously a hypothesis to explain its mechanism. In the present study, we substitute a synthetic auditory ossicle (Apaceram) for the bone to simplify the model system of experiments for our hypothesis. Its process was studied on a model simulation in vitro, in vivo and clinically at molecular level, using laser-Raman spectrometry. An increase of conductivity in both the saline solution and double-distilled water immersed Apaceram indicated demineralization depending on time lapse. The process of demineralization was revealed by the Raman spectral profiles as shown by narrowing a half-peak breadth (p < 0.01) of v1 signal (PO4(3-), 960 cm-1) and the disappearance of the shoulder at circa 950 cm-1 on the Apaceram surface implanted for 6 months. On the other hand, the process of remineralization was revealed in vivo by the spectral profiles: 1) a broader half-peak breadth (p < 0.01) of v1 signal on the Apaceram surface implanted for 10 months than that implanted for 6 months; 2) a weak reappearance of the shoulder at ca. 950 cm-1 on the Apaceram surface after 10 months; 3) four signals (v1 through v4) of PO4(3-) on the HOAP observed for inside the Apaceram shaft and 4) a weak v1 signal on both the granular fluorescent substance in the clinical case and the high density area of subcutaneous tissue after contact with the Apaceram in rats for 3 months. Judging from these investigations, it is proposed that the mechanism of bone destruction associated with cholesteatoma is a form of de- and remineralization.