Clinical Significance of Cancer Stem Cell Markers in Primary and Metastatic Tissues in Patients With Breast Cancer.

BACKGROUND/AIM: This study aimed to examine the clinical significance of the protein expression of the cancer stem cell (CSC) markers ALDH1A1, CD133, CD44, and MSI-1 in primary and metastatic tissues of patients with breast cancer (BC). PATIENTS AND METHODS: ALDH1A1, CD133, CD44, and MSI-1 protein expression in pairs of primary and metastatic tissues of 55 patients with BC with metastases treated at Kanagawa Cancer Center between January 1970 and December 2016 were evaluated using immunohistochemical assay and their association with clinicopathological factors and survival was examined. RESULTS: There were no significant differences in CSC marker expression rates between primary and metastatic tissues for any CSC markers. Regarding the relationship between CSC marker expression in primary tissues and survival, patients with high CD133 expression had significantly lower recurrence-free survival (DFS) and overall survival. On multivariate analysis, they were also a poor independent predictor of DFS (hazard ratio=4.993, 95%CI=2.189-11.394, p=0.0001). In contrast, there was no significant association between the expression of any CSC marker in metastatic tissues and survival. CONCLUSION: CD133 expression in the primary BC tissue may be a useful risk factor for recurrence in patients with BC.

Recurrence rate of intramucosal gastric cancer with positive vertical margin due to lesion damage during endoscopic submucosal dissection.

BACKGROUND AND STUDY AIM: In principle, additional surgery is performed after endoscopic submucosal dissection for early gastric cancer if the vertical margin is positive, regardless of lesion damage. The recurrence rate of vertical margin-positive lesions due to lesion damage after endoscopic submucosal dissection is unknown, and unnecessary surgeries may be performed. In this study, we investigated whether there was a difference in the recurrence rate between vertical margin-positive lesions due to lesion damage and vertical margin-negative lesions. PATIENTS AND METHODS: We included 1,294 intramucosal gastric cancer lesions that were resected by endoscopic submucosal dissection between January 2008 and December 2016, without additional surgery. The lesions were divided into the Damage and No damage groups based on vertical margin status. The Damage group had only one non-curative indication: a positive vertical margin due to lesion damage. The No damage group had no non curative indications. We compared the recurrence rate between the Damage and No damage groups. RESULTS: The recurrence rates of the Damage and No damage groups were 0% (0/23; 95% confidence interval: 0-14.8%) and 0% (0/1,271; 95% confidence interval: 0-0.003%), respectively, with no statistically significant difference. CONCLUSIONS: In intramucosal gastric cancer, the recurrence rate of vertical margin-positive lesions due to lesion damage was 0%, which did not differ from that of vertical margin-negative lesions with curative resection. Follow-up, instead of additional surgery, may be an option for patients with non-curative resection when the only non-curative indication is a positive vertical margin due to lesion damage.

Clinical and pathological aspects of gossypiboma in a dog: case report / Aspectos clínico-patológicos do gossipiboma em cão: relato de caso

A female dog was treated at the Veterinary Hospital Governador Laudo Natel for symptomscaused by an intra-abdominal gossypiboma (gossypium: cotton; boma: place of hiding). Showing high morbidity and mortality, the gossypiboma is a granulomatous reaction that is formed in response to exposure to a textile matrix. The objective of this report is to describe the clinical and pathological findings of an intra-abdominal gossypiboma in a female dog. This patient arrived at urgent care showing increased abdominal volume, pain, and cachexia. Ultrasonography showed a regular contour formation that could not be completely delimited. The patient underwent exploratory laparotomy and died in the postoperative period. The gossypiboma had a smooth, firm, and purple-gray exterior surface. Microscopy revealed a severe reaction of desmoplasia around the necrotic region, residual textile material, and adipose tissue. Negligence by veterinarians leading to this condition can result in patients' death as well as causing psychological damage to owners.(AU)

Uma cadela foi atendida no Hospital Veterinário "Governador Laudo Natel", apresentando sinais clínicos oriundos de gossipiboma intra-abdominal (gossypium: algodão e boma: local de ocultação), tais como aumento de volume abdominal, regurgitação e caquexia. A ultrassonografia evidenciou formação de contorno regular que não podia ser totalmente delimitada. A paciente foi submetida à laparotomia exploratória e veio a óbito no período pós-operatório. O gossipiboma mostrou-se com superfície externa lisa, firme e roxo-acinzentada apresentando-se à microscopia como reação severa de desmoplasia ao redor de material necrótico, resíduos de material têxtil e do tecido adiposo. A negligência por parte dos médicos veterinários pode levar pacientes ao óbito, bem como trazer prejuízos psicológicos aos proprietários. Objetiva-se, com este relato, descrever os achados clínicos e patológicos de uma cadela que apresentou um gossipiboma intra-abdominal.(AU)

Versatility of HDA19-deficiency in increasing the tolerance of Arabidopsis to different environmental stresses.

Histone acetylation is controlled by HATs and HDACs, which are essential epigenetic elements that regulate plant response to environmental stresses. A previous study revealed that a deficiency in an HDAC isoform (HDA19) increases tolerance to high salinity stress in the Arabidopsis wild-type Col-0 background. Here, the increased tolerance of hda19 to drought and heat stresses is demonstrated. Results indicate that hda19 plants have greater tolerance than wild-type plants to stress conditions. The data indicate that the stress response pathway coordinated by HDA19 plays a pivotal role in increasing tolerance to a variety of different abiotic stresses in Arabidopsis, including salinity, drought, and heat. The greater level of tolerance of hda19 plants to several different environmental stresses suggests that HDA19 represents a promising target for pharmacological manipulation in order to enhance abiotic stress tolerance in plants. ABBREVIATIONS: HAT, histone acetyltransferase; HDAC, histone deacetylase; HSF, heat shock transcription factor; RPD3, reduced potassium dependency 3; SIRT, Silent Information Regulator 2.

A importância dos exames neurológico e radiográfico no diagnóstico de lesões cervicais em equino: relato de caso / Fundamentals of neurological and radiographic examination on diagnosis of cervical lesions of a horse: case report

As lesões vertebrais cervicais são relativamente comuns nos equinos. No entanto, são poucas as descrições a respeito desse tema na literatura. Acredita-se que esse fato se deva, em parte, às limitações relacionadas ao diagnóstico. Em virtude de um traumatismo cervical, o animal pode apresentar sinais clínicos de paresia e paraplegia até tetraparesia ou tetraplegia, que podem culminar em estado comatoso e, eventualmente, em morte. Esses sinais clínicos podem ser detectados no exame físico, porém, quando associado à imaginologia (avaliação radiográfica), permite a obtenção de diagnóstico mais preciso da lesão. Consequentemente, um prognóstico mais adequado pode ser estabelecido para o paciente. Em face do exposto, apresenta-se o caso de uma égua Mangalarga, de quatro anos de idade, com sinais de tetraplegia pós-trauma, aumento de tônus muscular e paralisia espástica, com hiperreflexia, nos quatros membros. Por meio dos exames neurológicos realizados na paciente, em associação com as imagens radiográficas convencionais e contrastadas, foi possível estabelecer o diagnóstico do quadro. O animal apresentava lesões envolvendo as duas primeiras vértebras cervicais, atlas (C1) e áxis (C2), e a quinta e a sexta vértebra (C5-C6), destacando fratura de C1, deslocamento do dente de C2 e subluxação entre C5 e C6, com compressão ventrodorsal da medula espinhal. Confirmadas as suspeitas neurológicas e mediante prognóstico desfavorável, o animal foi submetido à eutanásia, com consentimento do proprietário. As lesões observadas nas imagens radiográficas foram confrontadas e confirmadas na necropsia.(AU)

Cervical vertebral lesions are relatively common in equine patients, but its diagnosis seems challenging for the clinician. Clinical signs most commonly associated with this clinical feature are paresis, paraplegia, tetraparesis, tetraplegia, which may lead the patient to a comatose state, and even death. Fortunately, these clinical signs can be perceived during physical examination. All of which, alongside imaging techniques, may allow a precise diagnose of the location of lesions and prediction of a prognosis for the patient. We present a case report of a Mangalarga mare, with four years of age, presenting signs of post-traumatic tetraplegia with spastic paralysis with increased muscular tonus and hyperreflexia in all four limbs. A thorough neurological examination of the patient associated with conventional and contrast medium radiographic imaging of the cervical region identified lesions at the atlas (C1) and axis (C2, and fifth (C5) and sixth (C6) cervical vertebrae. Fracture of the atlas vertebrae, dislocation of the axis' dens as well as subluxation of C5-C6 causing ventrodorsal medullary compression of the spinal cord were diagnosed radiographically. Due to the extent of the lesions the animal was euthanatized with the owner's consent. All lesions identified radiographically were posteriorly confirmed during post-mortem examination.(AU)

Vascular density of superficial esophageal squamous cell carcinoma determined by direct observation of resected specimen using narrow band imaging with magnifying endoscopy.

Observation of the microvasculature using narrow band imaging (NBI) with magnifying endoscopy is useful for diagnosing superficial squamous cell carcinoma. Increased vascular density is indicative of cancer, but not many studies have reported differences between cancerous and noncancerous areas based on an objective comparison. We observed specimens of endoscopic submucosal dissection (ESD) using NBI magnification, and determined the vascular density of cancerous and noncancerous areas. A total of 25 lesions of esophageal squamous cell carcinoma that were dissected en bloc by ESD between July 2013 and December 2013 were subjected to NBI magnification. We constructed a device that holds an endoscope and precisely controls the movement along the vertical axis in order to observe submerged specimens by NBI magnification. NBI image files of both cancerous (pathologically determined invasion depth, m1/2) and surrounding noncancerous areas were created and subjected to vascular density assessment by two endoscopists who were blinded to clinical information. The invasion depth was m1/2 in 20, m3/sm1 in four and sm2 in one esophageal cancer lesion. Mean vascular density was significantly increased in cancerous areas (37.6 ± 16.3 vessels/mm2) compared with noncancerous areas (17.6 ± 10.0 vessels/mm2) (P < 0.05). The correlation coefficients between vascular density determined by two endoscopists were 0.86 and 0.81 in cancerous and noncancerous areas, respectively. Receiver operating curve (ROC) analysis revealed that the area under the curve (AUC) of vascular density was 0.895 (95% CI, 0.804-0.986). For this ROC curve, sensitivity was 78.3% and specificity was 87.0% when the cutoff value of vascular density was 26 vessels/mm2. NBI magnification confirmed significant increases in vascular density in cancerous areas compared with noncancerous areas in esophageal squamous cell carcinoma. The rates of agreement between vascular density values determined by two independent operators were high.

Streptococcus thermophilus fermented milk reduces serum MDA-LDL and blood pressure in healthy and mildly hypercholesterolaemic adults.

Low-density lipoprotein (LDL)-cholesterol, malondialdehyde-modified low-density lipoprotein (MDA-LDL), MDA-LDL/LDL-cholesterol in serum, and blood pressure are considered useful risk markers of cardiovascular diseases. This study aimed to examine whether a fermented milk containing Streptococcus thermophilus YIT 2001 (ST), which has high anti-oxidative activity, would benefit healthy and mildly hyper-LDL-cholesterolaemic adults via a randomised, double-blind, placebo-controlled trial. ST-fermented milk or non-fermented placebo milk (PC) was consumed once a day for 12 weeks by 29 and 30 subjects, respectively, with average serum LDL-cholesterol levels of about 140 mg/dl. Serum levels of LDL-cholesterol and MDA-LDL and blood pressure were analysed before (baseline) and after consumption. Comparisons of the responses between both groups were assessed using analysis of covariance (ANCOVA, with the baseline value as the covariate). ANCOVA demonstrated that the ST group had significant reductions in MDA-LDL, MDA-LDL/LDL-cholesterol, systolic blood pressure (SBP), and diastolic blood pressure (DBP) compared with the PC group during the consumption period (P<0.05). Moreover, stratified analysis revealed that there were significant reductions in MDA-LDL, MDA-LDL/LDL-cholesterol, SBP, and DBP in the ST group compared with the PC group during the consumption period in subjects who had above median (65 U/l) levels of oxidative stress marker MDA-LDL at baseline (P<0.05), but not in subjects with levels below the median. These findings suggest that daily consumption of ST-fermented milk may be beneficial in healthy or mildly hyper-LDL cholesterolaemic subjects through reductions in risk marker values of oxidative stress and/or cardiovascular diseases. The benefits were particularly remarkable in subjects who had higher levels of MDA-LDL.

Empirical growth curve estimation considering multiple seasonal compensatory growths of body weights in Japanese Thoroughbred colts and fillies.

Thoroughbred horses are seasonal mating animals, and their foals are born yearly in spring seasons. In northern regions or countries, the foals generally show a typical seasonal compensatory growth pattern, where their growth rate declines in winter and increases in the next spring. In this study, a new empirical approach is proposed to adjust for this compensatory growth when growth curve equations are estimated, by using BW of Japanese Thoroughbred colts and fillies raised in Hidaka, Hokkaido. Based on the traditional Richards growth curve equation, new growth curve equations were developed and fit to the weight-age data. The foals generally experience 2 major winter seasons before their debut in horseracing. The new equations had sigmoid subfunctions that can empirically adjust the first and second year compensatory growths, combined with the Richards biological parameter responsible for the maturity of animals. The unknown parameters included in the equations were estimated by SAS NLMIXED procedure. The goodness-of-fit was examined by using several indices of goodness-of-fit (i.e., Akaike's information criterion, Bayesian information criterion, -2 log likelihood, and residual sum of squares) for the multiple applications of the subfunctions. The indices indicated the best fit of the new equations including both subfunctions for the first and second compensatory growths to the weight-age data. The shapes of the growth curves were improved during the periods of compensatory growth. The proposed method is one of the useful approaches for adjusting multiple seasonal compensatory growths in growth curve estimations of Thoroughbreds and for the management of young horses during the compensatory periods.

Octacalcium phosphate collagen composites with titanium mesh facilitate alveolar augmentation in canine mandibular bone defects.

This study was designed to investigate whether bone regeneration by implantation of octacalcium phosphate and porcine atelocollagen composite (OCP/Col) would be enhanced if mechanical stress to the implanted OCP/Col were alleviated. OCP/Col discs were implanted into an arc-shaped mandibular defect in male adult beagle dogs divided into untreated, OCP/Col, and OCP/Col/Mesh groups. In the OCP/Col/Mesh group, mechanical stress towards the implanted OCP/Col was alleviated by a titanium mesh. Bone regeneration in the three groups was compared after 6 months. Macroscopically, the alveolus in the OCP/Col/Mesh group was augmented vertically more than in the other two groups. Morphometric analysis by micro-CT showed the bone volume in the OCP/Col/Mesh group was significantly greater than in the other two groups. The augmented alveolus in the OCP/Col/Mesh group consisted of outer cortical and inner cancellous structure. Histologically, the OCP/Col/Mesh-treated alveolus was augmented by matured bone tissue along the inside of the titanium mesh. The implanted OCP/Col in the OCP/Col/Mesh and OCP/Col groups had almost disappeared. These results indicated that vertical bone regeneration by OCP/Col was efficient and successful when the mechanical stress to the implanted OCP/Col was alleviated. OCP/Col should be a useful bone substitute with active structural reconstitution.

Pharmacokinetics, safety, and tolerability of atomoxetine and effect of CYP2D6*10/*10 genotype in healthy Japanese men.

Atomoxetine is a cytochrome P4502D6 (CYP2D6) substrate. The reduced-activity CYP2D6*10 allele is particularly prevalent in the Japanese population and may contribute to known ethnic differences in CYP2D6 metabolic capacity. The purpose of this study was to examine atomoxetine pharmacokinetics, safety, tolerability, and the effect of the CYP2D6*10/*10 genotype after single-stepped dosing (10, 40, 90, or 120 mg) and at steady state (40 or 60 mg twice a day for 7 days) in 49 healthy Japanese adult men. Dose proportionality was shown and tolerability confirmed at all doses studied. Comparison of pharmacokinetics, safety, and tolerability between Japanese and US subjects showed no clinically meaningful ethnic differences. The CYP2D6*10/*10 subjects had 2.1- to 2.2-fold and 1.8-fold higher area under the plasma concentration-time curve values relative to the CYP2D6*1/*1 and *1/*2 subjects and the CYP2D6*1/*10 and *2/*10 subjects, respectively. The adverse events reported by CYP2D6*10/*10 subjects were indistinguishable from those of other Japanese participants. The higher mean exposure in CYP2D6*10/*10 subjects is not expected to be clinically significant.

Predicting the survival of experimental ischaemic small bowel using intraoperative near-infrared fluorescence angiography.

BACKGROUND: Predicting the long-term viability of ischaemic bowel during surgery is challenging. The aim was to determine whether intraoperative near-infrared angiography (NIR-AG) of ischaemic bowel might provide metrics that were predictive of long-term outcome. METHODS: NIR-AG using indocyanine green was performed on 24 pigs before, and after inducing bowel ischaemia to determine the feasibility of NIR-AG for detecting compromised perfusion. Contrast-to-background ratio (CBR) over time was measured in regions of interest throughout the bowel, and various metrics of the CBR-time curve were developed. Sixty rat small bowels, with or without strangulation, were imaged during surgery and on day 3 after operation. CBR metrics and clinical findings were assessed quantitatively for their ability to predict animal survival, histological grade of ischaemic injury and visible necrosis on day 3. RESULTS: In ischaemic pig small bowel, various qualitative and quantitative CBR metrics appeared to correlate with bowel injury as a function of distance from normal bowel. In rats, intraoperative clinical assessment showed high specificity but low sensitivity for predicting outcome on day 3 after operation. Qualitative patterns of the CBR-time curve, such as absence of an arterial inflow peak and presence of a NIR filling defect, resulted in better prediction of survival (90 per cent), histological grade (85 per cent) and visible necrosis on day 3 (92 per cent). CONCLUSION: Survival of ischaemic bowel was predicted by intraoperative NIR-AG with greater accuracy than clinical evaluation alone.

Comparison of net anaerobic energy utilisation estimated by plasma lactate accumulation rate and accumulated oxygen deficit in Thoroughbred horses.

REASONS FOR PERFORMING STUDY: Accumulated O(2) deficit (AOD) and plasma lactate accumulation rate (PLAR) are alternative methods for estimating net anaerobic energy utilisation (NAEU) in exercising horses. How they compare or their accuracy is unknown. OBJECTIVES: We hypothesised net anaerobic energy utilisation calculated by PLAR (NAUE(PLAR)) is equivalent to NAUE estimated by AOD (NAUE(AOD)). METHODS: Six Thoroughbred horses ran at identical supramaximal speeds (118% aerobic capacity) until exhaustion for 2 runs while breathing normoxic (NO, 21% O(2)) or hyperoxic (HO, 26% O(2)) gas. Jugular blood was sampled at 15 s intervals to measure plasma lactate concentration. Horses also ran at incremental submaximal speeds from 1.7-11.0 m/s to determine the linear relationship between speed and O(2) consumption to estimate O(2) demand for AOD calculations. RESULTS: Maximum O(2) consumption of horses increased 11.6 ± 2.3% in HO and NAEU(PLAR) and NAUE(AOD) decreased 38.5 ± 8.0% and 46.2 ± 17.7%, respectively. The NAEU(PLAR) in NO was 114.5 ± 27.4 mlO(2) (STPD) equivalent/kg bwt contributing 23.5 ± 3.7% to total energy turnover and in HO was 70.9 ± 19.8 mlO(2) (STPD) equivalent/kg bwt contributing 14.6 ± 3.8% to total energy turnover. The NAUE(AOD) in NO was 88.6 ± 24.3 mlO(2) (STPD) equivalent/kg bwt contributing 19.9 ± 2.1% to total energy turnover and in HO was 56.2 ± 19.1 mlO(2) (STPD) equivalent/kg bwt contributing 10.9 ± 4.3% to total energy turnover. Overall, NAEU(AOD) was systematically biased -23.5 ± 16.8 mlO(2) (STPD) equivalent/kg bwt below NAEU(PLAR). Total energy demand estimated by PLAR was 11.1 ± 5.4% greater than that estimated by AOD and was higher in every horse. CONCLUSIONS: The NAUE(PLAR) estimates average 40.0 ± 29.6% higher than NAUE(AOD) and are highly correlated (r(2) = 0.734), indicating both indices are sensitive to similar changes in NAEU. Accuracy of the estimates remains to be determined. Multiple considerations suggest NAUE(AOD) may underestimate total energy cost during high-speed galloping, thus biasing low the AOD estimate of NAEU.

Bone regeneration by octacalcium phosphate collagen composites in a dog alveolar cleft model.

Octacalcium phosphate (OCP) and porcine atelocollagen sponge composites (OCP/Col) markedly enhanced bone regeneration in a rat cranial defect model. To assess clinical application, the authors examined whether OCP/Col would enhance bone regeneration in an alveolar cleft model in an adult dog, which was assumed to reflect patients with alveolar cleft. Disks of OCP/Col or collagen were implanted into the defect and bone regeneration by OCP/Col or collagen was investigated 4 months after implantation. Macroscopically, the OCP/Col-treated alveolus was obviously augmented and occupied by radio-opacity, and the border between the original bone and the defect was indistinguishable. Histological analysis revealed it was filled and bridged with newly formed bone; a small quantity of the remaining implanted OCP was observed. X-ray diffraction patterns of the area of implanted OCP/Col indicated no difference from those of dog bone. In the collagen-treated alveolus, the hollowed alveolus was mainly filled with fibrous connective tissue, and a small amount of new bone was observed at the defect margin. These results suggest that bone was obviously repaired when OCP/Col was implanted into the alveolar cleft model in a dog, and OCP/Col would be a significant bone regenerative material to substitute for autogeneous bone.

Intra-bone marrow cotransplantation of donor mesenchymal stem cells in pig-to-NOD/SCID mouse bone marrow transplantation facilitates short-term xenogeneic hematopoietic engraftment.

We directly injected porcine donor mesenchymal stem cells (MSC) into murine bone marrow (BM) cavities to examine the effects of intra-BM cotransplantation of MSC in pig-to-NOD/SCID mouse bone marrow transplantation (BMT) on xenogeneic engraftment. Porcine MSC prepared by aspiration of iliac BM of miniature swine were identified as CD90+CD29+CD45-CD31- and shown to differentiate into osteoblastocytes and adipocytes. A few weeks after expansion, MSC (1 x 10(6) cells/mouse) were directly injected with BM cells (30 x 10(6) cells/mouse) obtained from vertebrae through a microsyringe into BM cavities of both tibiae of NOD/SCID mice after 3-Gy total body irradiation. Controls were injected with only BM cells. Porcine chimerisms of BM cells of tibiae (injection site) and of femurs (non-injection site) in recipient mice were evaluated with porcine and murine cell markers using FACS. The chimerism of porcine class I+ cells at the injection site in the MSC group and the controls were 3.45%, 1.43%, and 0.17%, and 2.27%, 0.81%, and 0.1% at 1, 3, and 6 weeks, respectively. The chimerism at the noninjection site in the MSC group and the controls were 0.21%, 1.34%, and 0.11%, and 0.06%, 0.42%, and 0.09% at 1, 3, and 6 weeks, respectively. The total chimerisms of injection site in the MSC group to 6 weeks were significantly higher than those in the control group (1.60% vs 0.99%; P < .05), whereas the chimerism of the noninjection site in MSC group was remarkably higher at 3 weeks. In conclusion, intra-BM cotransplantation of porcine donor MSC in pig-to-NOD/SCID mouse BMT improved short-term xenogeneic engraftment, presumably due to humoral factors.

Dysphagia-gastroesophageal reflux complex: complications due to dysfunction of solitary tract nucleus-mediated vago-vagal reflex.

We report on the complication of gastroesophageal reflux (GER) in four patients with lower brainstem dysfunction. These patients suffered from perinatal asphyxia, cerebellar hemorrhage, or congenital dysphagia of unknown origin and showed facial nerve palsy, inspiratory stridor due to vocal cord paralysis, central sleep apnea, and dysphagia, in various combinations. Naso-intestinal tube feeding was introduced in all of the patients due to recurrent vomiting and aspiration pneumonia resulting from GER. T2-weighted magnetic resonance (MR) imaging revealed symmetrical high intensity lesions in the tegmentum of the lower pons and the medulla oblongata in two of the patients, and pontomedullary atrophy in another patient. In normal subjects, lower esophageal sphincter contraction is provoked by distension of the gastric wall, through a vago-vagal reflex. Since this reflex arc involves the solitary tract nucleus, where the swallowing center is located, the association of dysphagia and GER in the present patients is thought to result from the lesions in the tegmentum of medulla oblongata. We propose the term "dysphagia-GER complex" to describe the disturbed motility of the upper digestive tract due to lower brainstem involvement. In children with brainstem lesions, neurological assessment of GER is warranted, in addition to the examination of other signs of brainstem dysfunction, including dysphagia and respiratory disturbance.

The serum levels of proinsulin and their relationship with IGFBP-1 in obese children.

AIM: Serum proinsulin (PI) levels were investigated in obese children to determine whether PI is a sensitive indicator of insulin resistance, as previously shown in adults with type 2 diabetes mellitus (DM), and to evaluate their relationship with insulin-like growth factor-binding protein-1 (IGFBP-1) known as a predictor of the development of cardiovascular disease in diabetic adults. SUBJECTS AND METHODS: Forty-two obese children without DM (age, 12.1 +/- 1.5 year) and 42 age-matched control children were included in the study. The serum levels of PI, immunoreactive insulin (IRI), IGFBP-1 and free insulin-like growth factor-1 (IGF-1) were measured in the fasting state. RESULTS: The fasting levels of serum PI and IRI were significantly higher in obese children than in controls (PI, 10.5 +/- 6.8 vs. 5.6 +/- 2.0 pmol/l, p < 0.001; IRI, 72.0 +/- 41.8 vs. 32.7 +/- 19.5 pmol/l, p < 0.001). Serum IGFBP-1 levels were significantly lower in obese children than in controls (37.7 +/- 24.6 vs. 76.3 +/- 26.5 microg/l, p < 0.001). The ratio of PI to IRI (calculated as molar ratios) did not differ significantly between obese and control subjects (0.16 +/- 0.08 vs. 0.19 +/- 0.11, p = 0.08). For the whole group, serum PI levels correlated positively with IRI and inversely with IGFBP-1 (IRI, r = 0.67, p < 0.001; IGFBP-1, r = -0.49, p < 0.001). Serum IGFBP-1 levels correlated inversely with both BMI and IRI (BMI, r = -0.73, p < 0.001; IRI, r = -0.60, p < 0.001). Multiple regression analysis revealed that the best predictive parameters for IGFBP-1 were BMI and PI (R2 = 0.57, p < 0.001 and p < 0.05, respectively). CONCLUSION: These findings suggest that fasting serum PI levels may be a better predictor than fasting insulin levels for the future development of type 2 DM and cardiovascular disease in obese children, and PI, in addition to insulin, contributes to the suppression of hepatic IGFBP-1 production.

Effect of amino acid and glucose administration following exercise on the turnover of muscle protein in the hindlimb femoral region of thoroughbreds.

REASONS FOR PERFORMING STUDY: In man, muscle protein synthesis is accelerated by administering amino acids (AA) and glucose (Glu), because increased availability of amino acids and increased insulin secretion, is known to have a protein anabolic effect. However, in the horse, the effect on muscle hypertrophy of such nutrition management following exercise is unknown. OBJECTIVES: To determine the effect of AA and Glu administration following exercise on muscle protein turnover in horses. We hypothesise that administration of AA and Glu after exercise effects muscle hypertrophy in horses, as already shown in man and other animals. METHODS: Measurements of the rate of synthesis (Rs) and rate of degradation (Rd) of muscle protein in the hindlimb femoral region of thoroughbred horses were conducted using the isotope dilution method to assess the differences between the artery and iliac vein. Six adult Thoroughbreds received a continuous infusion of L-[ring-2H5]- phenylalanine during the study, the stable period for plasma isotope concentrations (60 min), resting periods (60 min), treadmill exercise (15 min) and recovery period (240 min). All horses were given 4 solutions (saline [Cont], 10% AA [10-AA], 10% Glu [10-Glu] and a mixture with 10% AA and 10% Glu [10-Mix]) over 120 min after exercise, and the Rs and Rd of muscle protein in the hindlimb measured. RESULTS: The average Rs during the 75-120 min following administration of 10-Mix was significantly greater than for the other solutions (P<0.05). The second most effective solution was 10-AA, and there was no change in Rs after 10-Glu. CONCLUSIONS: Administration of AA following exercise accelerated Rs in the hindlimb femoral region, and this effect was enhanced when combined with glucose, because of increasing insulin secretion or a decreased requirement for AA for energy. POTENTIAL RELEVANCE: Further studies are required regarding the effect on muscle hypertrophy of supplementing amino acids and glucose in the feed of exercising horses.

Haplotype analysis of a 100 kb region spanning TNF-LTA identifies a polymorphism in the LTA promoter region that is associated with atopic asthma susceptibility in Japan.

BACKGROUND: The tumour necrosis factor (TNF) gene family, which includes TNF, LTA, and LTB, is located consecutively on human chromosome 6p21 region, which has been linked to asthma by several genome-wide screens. (LTA, lymphotoxin-alpha; LTB, lymphotoxin-beta). OBJECTIVE: The aim of the present study was to determine whether genes on 6q21 are related to development of atopic asthma. Methods We screened for mutations in the coding and promoter regions of genes in the TNF-LTA region, including BAT1, NFKBIL1, LTA, TNF, LTB, AIF, and BAT2, and conducted a transmission disequilibrium test of 41 polymorphisms in 137 families identified through pro-bands with childhood-onset atopic asthma. (BAT1, HLA-B-associated transcript 1; NFKBIL1, nuclear factor of kappa light polypeptide gene enhancer in B cells inhibitor-like 1; AIF, allograft inflammatory factor 1). RESULTS: Haplotypes of the LTA/TNF linkage disequilibrium block were associated significantly with asthma (global P=0.0097). Transmission patterns of the common haplotypes to asthmatic offspring were predicted by a single-nucleotide polymorphism in the LTA promoter region. The G allele of the LTA-753G/A polymorphism was transmitted preferentially to asthma-affected individuals (P=0.001). Luciferase reporter assays with constructs containing the 5' and 3' flanking regions of the LTA gene showed 30-50% lower transcriptional activity when the -753A allele was present than that of other haplotypes. CONCLUSION: Our results suggest that LTA is one of the genes that contributes to susceptibility to atopic asthma, and that the association of the TNF/LTA haplotypes to asthma may be defined by the polymorphism in the LTA promoter region in the Japanese population.