Masticatory performance-related factors in preschool children: establishing a method to assess masticatory performance in preschool children using colour-changeable chewing gum.

A knowledge gap regarding masticatory performance in preschool children exists, which in turn delays intervention for preventive care; therefore, a method to easily assess performance is needed. The purpose of this study is to investigate the feasibility of assessing masticatory performance using colour-changeable chewing gum and to investigate masticatory performance-related factors in preschool children. This cross-sectional survey was conducted in two childcare facilities and our laboratory. First, a one-third quantity of colour-changeable chewing gum was masticated by six adults to assess the nature and progression of colour changes in this quantity. Then, masticatory performance in 370 children 4-6 years of age was assessed using the same quantity of colour-changeable chewing gum (60 chew strokes). The maximum bite force, body height, weight, age and number of healthy teeth were recorded. A t-test was performed to determine whether gum-chewing experience or lack thereof produced a significant difference in masticatory performance. The Spearman's rank correlation coefficient was then determined for masticatory performance assessment values and other factors solely for children with gum-chewing experience. Measurements from 259 children were obtained. Children with gum-chewing experience demons trated significantly higher assessment values and were deemed to have been correctly assessed. A very weak but significant positive correlation was observed only between masticatory performance and the number of healthy teeth. The masticatory performance of preschool children was easily assessed using colour-changeable chewing gum. The assessment values demonstrated significant correlation with the number of healthy teeth, but not with maximum bite force, body height, weight or age.

Rapid Communication: MiR-92a as a housekeeping gene for analysis of bovine mastitis-related microRNA in milk.

Our aim was to identify a suitable microRNA housekeeping gene for real-time PCR analysis of bovine mastitis-related microRNA in milk. We identified , , and as housekeeping gene candidates on the basis of previous Solexa sequencing results. Threshold cycle (CT) values for , , and did not differ between milk from control cows and milk from mastitis-affected cows. NormFinder software identified as the most stable single housekeeping gene. We evaluated the suitability of the housekeeping gene candidates by using them to assess expression levels of the inflammation-related gene . Regardless of the housekeeping gene candidates used for normalization, relative expression levels of were significantly higher in mastitis-affected samples than in control samples. However, of all the housekeeping genes and gene combinations investigated, normalization with alone generated the difference in relative expression between mastitis-affected and control samples with the highest significance. These results suggest that is suitable for use as a housekeeping gene for analysis of bovine mastitis-related microRNA in milk.

Efficacy of a combined protocol for re-insemination of open cows after early pregnancy diagnosis using ultrasonography and its effect on fertility.

The objective of the present field study was to establish the beneficial effects of re-insemination of non-pregnant cows using ultrasonography 20 to 23 days after the artificial insemination. A total of 245 Japanese Black cows were artificially inseminated and early pregnancy diagnosis (EPD) was performed on 92 cows 20 days after insemination, using ultrasonography. The remaining 153 cows were considered as negative controls in which routine rectal palpation was performed for pregnancy diagnosis 45-50 days post-insemination. EPD revealed that eleven of the 92 cows (12%) were infertile due to ovarian abnormalities and were thus excluded from the rest of the study. Forty-eight (59%) of the remaining 81 cows were diagnosed as pregnant, while the other 33 (41%) were diagnosed as non-pregnant. Of these non-pregnant cows, 17 of them received a dose of an analogue of the gonadotropin-releasing hormone (GnRH analogue) and were then timed-inseminated, while the other 16 were observed for estrus signs, and 13 of them (81%) were artificially inseminated. Rates of conception were 35% and 38% in the GnRH and the artificially inseminated groups, respectively (P>0.05). Total pregnancy rate for the EPD group increased significantly (74%) (P<0.01) when compared to the control cows (54%) within the same period. In conclusion, our field study demonstrated that re-insemination of non-pregnant cows following EPD is highly efficacious not only in improving the rate of fertility via reducing inter-insemination and inter-calving intervals, but also aids in the early detection of ovarian disorders.

Comparison of oestrous intensity between natural oestrus and oestrus induced with ovsynch based treatments in Japanese black cows.

The aim of this study was to examine the possible differences of oestrous intensity between natural oestrus and induced oestrus using the walking activity measuring device. Walking activity was used as an evaluation index of oestrous intensity. A total of 27 Japanese Black cows, more than 40 days after calving and clinically normal, were randomly assigned to three groups. Walking activity was recorded using a commercially available computerized pedometer system. The treatment groups consisted of an Ovsynch (n = 8) and a controlled internal drugs releasing device (CIDR) + Ovsynch (n = 9) group. The control group (n = 10) received no treatment. Walking activity was examined in all groups. Timed artificial insemination (timed AI) was performed at 16 hours after the onset of oestrus in the control group and at 24 h after second administration of GnRH in the treatment groups. Duration of oestrus had a tendency to be shorter in both the Ovsynch and the CIDR + Ovsynch groups when compared with the control group. The time required from the onset of oestrus to the time showing the highest number of steps of walking (the time to peak) showed a tendency to be shorter in CIDR + Ovsynch group. The number of steps of walking at peak and overall walking activities were significantly lower in both treatment groups than in the control group. Both activity and super-activity periods of time in the treatment groups were shorter than the control group. No difference was observed in the conception rate between the control (50.0%; 10/20), Ovsynch (50.0%; 4/8) and CIDR + Ovsynch groups (66.7%; 6/9). This study demonstrates that the oestrous intensity of cows in oestrus was different between natural oestrus and induced oestrus and also between the methods of the synchronization, but no difference was observed in the conception rate among the three groups.

Role of interleukin-1beta and tumor necrosis factor-alpha-dependent expression of cyclooxygenase-2 mRNA in thermal hyperalgesia induced by chronic inflammation in mice.

The present study investigated whether the endogenous pro-inflammatory cytokines [interleukin (IL)-1beta and tumor necrosis factor-alpha (TNF-alpha)]-dependent expression of cyclooxygenase-2 (COX-2) mRNA within the spinal cord could be involved in the development of chronic inflammatory pain-like behaviors in mice. We demonstrated that the expression of COX-2 mRNA on the ipsilateral side of the spinal cord was significantly increased 6 h and 3 days after intraplantar injection of complete Freund's adjuvant (CFA), compared with the expression in saline-treated mice. In addition, the chronic pain-like behaviors following CFA injection were markedly suppressed by repeated intrathecal (i.t.) pre-treatment with the COX-2 inhibitor etodolac, but not with the COX-1 inhibitor mofezolac. The cytosolic level of the activated form of nuclear factor-kappa B (NF-kappaB), which is a major contributor to the induction of COX-2, on the ipsilateral side of the mouse spinal cord was also increased compared with that in the saline-treated mice. The key finding in the present study was that a single i.t. injection with either IL-1beta or TNF-alpha induced a marked increase in spinal COX-2 mRNA and persistent thermal hyperalgesia in mice. Furthermore, CFA-induced hypersensitivity to inflammatory pain was significantly reduced by repeated i.t. pre-injection of the recombinant Fc chimera of IL-1 receptor I or soluble TNF receptor I, which sequesters endogenous IL-1beta or TNF-alpha, respectively. In contrast, the expression of spinal COX-2 mRNA in CFA-treated mice was similar to that in saline-treated mice at 7 days after CFA injection. The present findings strongly indicate the early intrathecal use of the COX-2 inhibitor for the relief of chronic inflammatory pain. Furthermore, together with the result in a previous study that pro-inflammatory cytokines lead to stimulation of a NF-kappaB-dependent transcriptional pathway, these findings suggest that a spinal cytokine/NF-kappaB/COX-2 pathway may play an important role in the development, but not maintenance, of chronic pain following peripheral tissue inflammation.

Effect of hydrogen peroxide on quality of fresh-cut tomato.

The effect of hydrogen peroxide (H2O2; 0, 0.1, 0.2, and 0.4 M) on selected nutritional quality of fresh-cut tomato was investigated. Microbial population of tomato slices stored at 10 degrees C and treated with H2O2 was lower than the control by 1- (0.2 and 0.4 M) and 5-log (0.4 M), 3 and 7 d after processing, respectively. Dipping fresh-cut tomato into H2O2 resulted in reduced phenolic and antioxidant levels after 7 d in storage by at least 5% and 20%, respectively, and produced an initial decline in vitamin C and lycopene. Change in color values in the H2O2 treatments were associated with reduced carotenoid content. Our results confirmed antimicrobial benefits of H2O2 but revealed a compromise in antioxidant and carotenoid contents of fresh-cut tomatoes.

Nuclear reprogramming by somatic cell nuclear transfer--the cattle story.

Somatic cell nuclear transfer (cloning) returns a differentiated cell to a totipotent status; a process termed nuclear reprogramming. Nuclear transfer has potential applications in agriculture and biomedicine, but is limited by low efficiency. To understand the deficiencies of nuclear reprogramming, our research has focused on both candidate genes (imprinted and X-linked genes) and global gene expression patterns in cloned bovine embryos/offspring as compared to those generated by conventional reproduction. We found aberrant expression patterns of H19 and Igf2r as well as X-linked genes in term cloned calves. The expression profiles of cloned blastocysts, however, closely resembled those of the naturally fertilized embryos but were considerably different from those of their nuclear donor cells. Our findings suggest that cloned embryos have undergone significant nuclear reprogramming by the blastocyst stage. However, it is possible that during re-differentiation in later development gene expression aberrancies occur. Additionally, small initial nuclear reprogramming errors may be manifested during subsequent development.

Microsurgical excision of ganglioneuroma arising from the C8 nerve root within the neuroforamen.

Ganglioneuromas are benign, slow-growing tumors originating from sympathetic nerves or peripheral nerves, often associated with multiple tumor syndromes. They occasionally occur as spinal lesions and grow within the spinal canal or as paraspinal lesions. In this report, we describe a rare solitary ganglioneuroma arising from the cervical nerve root (C8) within the intervertebral foramen in adults. The tumor could be detected as a mass limited to the neuroforamen at an early stage by MR images. Unilateral microsurgical foraminotomy and EN BLOC resection of the tumor resulted in disappearance of the symptoms. Microsurgical resection of the relevant nerve root through limited medial foraminotomy at an intricate anatomical region of the cervico-thoracic junction was appropriate in the current case for complete resection of the tumor as well as to prevent postoperative structural weakness that could result in late segmental instability.

Cruciate paralysis and hemiplegia cruciata: report of three cases.

STUDY DESIGN: Report of three cases of cruciate paralysis and hemiplegia cruciata. OBJECTIVE: To stress the importance of upper cervical spine lesions causing neurological symptoms and signs. SETTING: Neuro-orthopedic service, Fukui University Hospital, Japan. RESULTS: Three patients (all females; one with congenital anomaly at the occiput-atlas level, one with assimilation of the atlas, and one with rheumatoid arthritis-related proliferative synovium) had clinical features of cruciate paralysis and hemiplegia cruciata. All three cases underwent decompressive surgeries. CONCLUSION: Neurological symptoms and signs of cruciate paralysis and hemiplegia cruciata should be carefully assessed, and surgical therapy should be based on the pathological condition.

Quantitative trait loci analysis for growth and carcass traits in a half-sib family of purebred Japanese Black (Wagyu) cattle.

We used a half-sib family of purebred Japanese Black (Wagyu) cattle to locate economically important quantitative trait loci. The family was composed of 348 fattened steers, 236 of which were genotyped for 342 microsatellite markers spanning 2,664 cM of 29 bovine autosomes. The genome scan revealed evidence of 15 significant QTL (<5% chromosome-wise level) affecting growth and carcass traits. Of the 15 QTL, six QTL were significant at the 5% experiment-wise level and were located in bovine chromosomes (BTA) 4, 5, and 14. We analyzed these three chromosomes in more detail in the 348 steers, with an average marker interval of 1.2 cM. The second scan revealed that the same haplotype of the BTA 4 region (52 to 67 cM) positively affected LM area and marbling. We confirmed the QTL for carcass yield estimate on BTA 5 in the region of 45 to 54 cM. Five growth-related QTL located on BTA 14, including slaughter and carcass weights, were positively affected by the same region of the haplotype of BTA 14 (29-51 cM). These data should provide a useful reference for further marker-assisted selection in the family and positional cloning research. The research indicates that progeny design with moderate genotyping efforts is a powerful method for detecting QTL in a purebred half-sib family.

Quantitation of bovine macrophage colony-stimulating factor in bovine serum by ELISA.

We established an enzyme-linked immunosorbent assay (ELISA) system for the quantitation of bovine macrophage colony-stimulating factor (M-CSF) and used it to measure the serum M-CSF levels in bovine fetuses and calves. The average serum M-CSF level was 2.7+/-1.5 ng/ml in 39 calves under 100 days old, and 1.8+/-0.8 ng/ml in 15 cattle between 101 and 418 days old. Fetal sera samples (n = 6) prepared from cattle between 150 and 280 days of gestational age had a higher average level of M-CSF (8.8+/-1.4 ng/ml). Alteration in serum M-CSF levels in each individual calf was also measured. The serum levels of M-CSF in calves at 0-1 day after birth ranged from 0.52 to 7.3 ng/ml. During the period 113-125 days after birth, serum levels were around 1.4+/-0.39 ng/ml. Although serum M-CSF levels generally decreased as the age of calves advanced, differences among individuals, especially among newborn calves, were observed.

Epigenetic characteristics and development of embryos cloned from donor cells treated by trichostatin A or 5-aza-2'-deoxycytidine.

Development to blastocyst following nuclear transfer is dependent on the donor cell's ability to reprogram its genome to that of a zygote. This reprogramming step is inefficient and may be dependent on a number of factors, including chromatin organization. Trichostatin A (TSA; 0-5 microM), a histone deacetylase inhibitor, was used to increase histone acetylation and 5-aza-2'-deoxycytidine (5-aza-dC; 0-5 microM), a DNA methyl-transferase inhibitor, was used to decrease methylation of chromatin in donor cells in an attempt to improve their reprogrammability. Adult fibroblast cells treated with 1.25 or 5 microM TSA had elevated histone H3 acetylation compared to untreated controls. Cells treated with 0.3 microM 5-aza-dC had decreased methylation compared to untreated controls. Both drugs at 0.08 microM caused morphological changes of the donor cells. Development to blastocysts by embryos cloned from donor cells after 0.08 or 0.3 microM 5-aza-dC treatments was lower than in embryos cloned from untreated control cells (9.7% and 4.2%, respectively, vs. 25.1%), whereas 0.08 microM TSA treatment of donor cells increased blastocyst development compared to controls (35.1% vs. 25.1%). These results indicate that partial erasure of preexisting epigenetic marks of donor cells improves subsequent in vitro development of cloned embryos.

Perturbation of estradiol-feedback control of luteinizing hormone secretion by immunoneutralization induces development of follicular cysts in cattle.

We used immunoneutralization of endogenous estradiol to investigate deficiencies in the estradiol-feedback regulation of LH secretion as a primary cause of follicular cysts in cattle. Twenty-one cows in the prostaglandin (PG) F(2alpha)-induced follicular phase were assigned to receive either 100 ml of estradiol antiserum produced in a castrated male goat (n = 11, immunized group) or the same amount of castrated male goat serum (n = 10, control group). The time of injection of the sera was designated as 0 h and Day 0. Five cows in each group were assigned to subgroups in which we determined the effects of estradiol immunization on LH secretion and follicular growth during the periovulatory period. The remaining six estradiol-immunized cows were subjected to long-term analyses of follicular growth and hormonal profiles, including evaluation of pulsatile secretion of LH. The remaining five control cows were used to determine pulsatile secretion of LH on Day 0 (follicular phase) and Day 14 (midluteal phase). The control cows exhibited a preovulatory LH surge within 48 h after injection of the control serum, followed by ovulation of the dominant follicle that had developed during the PGF(2alpha)-induced follicular phase. In contrast, the LH surge was not detected after treatment with estradiol antiserum. None of the 11 estradiol-immunized cows had ovulation of the dominant follicle, which had emerged before estradiol immunization and enlarged to more than 20 mm in diameter by Day 10. Long-term observation of the six immunized cows revealed that five had multiple follicular waves, with maximum follicular sizes of 20-45 mm at 10- to 30-day intervals for more than 50 days. The sixth cow experienced twin ovulations of the initial persistent follicles on Day 18. The LH pulse frequency in the five immunized cows that showed the long-term turnover of cystic follicles ranged from 0.81 +/- 0.13 to 0.97 +/- 0.09 pulses/h during the experiment, significantly (P < 0.05) higher than that in the midluteal phase of the control cows (0.23 +/- 0.07). The mean LH concentration in the immunized cows was also generally higher than that in the luteal phase of the control cows. However, the LH pulse and mean concentration of LH after immunization were similar to those in the follicular phase of the control cows. Plasma concentrations of total inhibin increased (P < 0.01) concomitant with the emergence of cystic follicles and remained high during the growth of cystic follicles, whereas FSH concentrations were inversely correlated with total inhibin concentrations. In conclusion, neutralization of endogenous estradiol resulted in suppression of the preovulatory LH surge but a normal range of basal LH secretion, and this circumstance led to an anovulatory situation similar to that observed with naturally occurring follicular cysts. These findings provide evidence that lack of LH surge because of dysfunction in the positive-feedback regulation of LH secretion by estradiol can be the initial factor inducing formation of follicular cysts.

Cloning of aged animals: a medical model for tissue and organ regeneration.

Cloning by nuclear transfer has great potential application in pharmaceutical protein production, xeno-transplantation, and perhaps most excitingly, therapeutic cloning. In therapeutic cloning a patient's own skin cells can be used to generate cloned embryos from which embryonic stem cells are isolated. Through targeted differentiation, embryonic stem cells can be directed to develop into the desired tissues/organs for replacement. The combination of homologous recombination of genes and nuclear transfer also offers the promise of correcting defective genes in humans. Demonstration of the successful cloning of aged animals is important for these future medical applications because degenerative diseases often afflict older adults. Our studies have demonstrated that skin fibroblast cells from aged adults, even after prolonged culture, provide nuclear donors equally as competent for cloning as cells from young adults or fetuses. These findings have paved the way for medically treating degenerative diseases of aged humans by tissue regeneration technologies made possible through cloning and homologous recombination.

Ligamentum flavum hematoma in the thoracic spine.

We report a case of a hematoma of ligamentum flavum at T11-12 in a 66-year-old man who presented with progressive weakness of the right foot and numbness of both legs. Past history was negative and no precipitating episode of lower back sprain or trauma. The resected T11 and T12 laminas showed old hematoma with degenerative changes in the ligamentum flavum. Hematoma occurring in the thoracic spine has never been reported previously.

Final height and growth hormone secretion after bone marrow transplantation in children.

Growth hormone (GH) deficiency has been regarded as a principal determinant for growth failure following bone marrow transplantation (BMT). We herein analyzed final height and GH secretion in the patients who received BMT during childhood. The study on final height in 30 patients (23 males; 19 with malignant disease) who underwent BMT before or at the onset of puberty showed the following findings: (1) Final height SD score (SDS) significantly decreased compared to pretreatment height SDS. (2) Patients who underwent BMT before the age of 10 years showed significantly greater reduction in height SDS compared to those who received after the age of 10 years. (3) The type of disease or a difference in preconditioning regimen did not influence the outcome of growth. (4) No patient showed GH deficiency. The study on GH secretion included 71 patients who had been followed for more than 5 years and who underwent insulin tolerance test more than twice following BMT. Thirteen patients experienced poor GH response at least once. Two of these patients had poor GH response repeatedly. In conclusion, children who undergo BMT at younger age have a higher risk of growth failure, and GH deficiency is not a major contributing factor for growth impairment following BMT.

Moyamoya disease complicated with renal artery stenosis and nephrotic syndrome: reversal of nephrotic syndrome after nephrectomy.

A 7-year-old boy with moyamoya disease developed sustained hypertension, nephrotic syndrome, hyperreninemia, and occlusion of the right renal artery. After right nephrectomy, hyperreninemia and hypertension improved. Proteinuria was resolved after nephrectomy, in parallel with the decrease in plasma renin activity. Moyamoya disease can cause nephrotic-range proteinuria, which is caused hemodynamically by hyperreninemia.