Egg laying performance and egg quality with Paracoccus carotinifaciens supplementation containing high astaxanthin levels.

1. This study assessed 1) the effects of Paracoccus carotinifaciens supplementation containing high astaxanthin levels on egg production performance and quality, 2) dynamics of carotenoids levels in the egg yolk and 3) taste of astaxanthin-rich egg yolk.2. Laying hens were fed diets containing different levels of P. carotinifaciens-derived astaxanthin (ASX; 0, 2, 4, 8, or 16 ppm) for 28 d (experiment 1) or a diet containing 16 ppm astaxanthin for 28 d followed by a 0 ppm astaxanthin diet for 28 days (experiment 2).3. Production performance, egg quality and egg yolk carotenoid levels were examined in experiment 1 (Ex1) and the dynamics of egg yolk carotenoid levels and egg yolk taste in experiment 2 (Ex2).4. ASX supplementation did not affect production performance or egg quality. ASX levels in the egg yolk became saturated after seven days of 16 ppm supplementation and decreased to less than one-tenth of the saturated levels seven days after supplementation cessation. Supplementation with 16 ppm ASX for 28 d did not affect egg yolk taste.5. Supplementation resulted in the production of ASX-rich eggs for a brief period without affecting production performance, egg quality or taste. Understanding the time taken for the incorporation of ASX into egg yolks is beneficial for value-added egg production and may help in minimising supplementation costs.

Analysis of signal components < 500 Hz in brain organoids coupled to microelectrode arrays: A reliable test-bed for preclinical seizure liability assessment of drugs and screening of antiepileptic drugs.

Brain organoids with three-dimensional structure and tissue-like function are highly demanded for brain disease research and drug evaluation. However, to our knowledge, methods for measuring and analyzing brain organoid function have not been developed yet. This study focused on the frequency components of an obtained waveform below 500 Hz using planner microelectrode array (MEA) and evaluated the response to the convulsants pentylenetetrazol (PTZ) and strychnine as well as the antiepileptic drugs (AEDs) perampanel and phenytoin. Sudden and persistent seizure-like firing was observed with PTZ administration, displaying a concentration-dependent periodic activity with the frequency component enhanced even in one oscillation characteristic. On the other hand, in the administration of AEDs, the frequency of oscillation decreased in a concentration-dependent manner and the intensity of the frequency component in one oscillation also decreased. Interestingly, at low doses of phenytoin, a group of synchronized bursts was formed, which was different from the response to the perampanel. Frequency components contained information on cerebral organoid function, and MEA was proven useful in predicting the seizure liability of drugs and evaluating the effect of AEDs with a different mechanism of action. In addition, frequency component analysis of brain organoids using MEA is an important analysis method to perform in vitro to in vivo extrapolation in the future, which will help explore the function of the organoid itself, study human brain developments, and treat various brain diseases.

Active transport of glucose across the jejunal epithelium decreases with age in broiler chickens.

Intestinal glucose absorption varies with growth; however, the dynamics of these variations has not been yet fully elucidated in broiler chickens. The present study aimed to compare jejunal glucose uptake and maltose digestion in broilers of 2 different ages, i.e., 1- vs. 5 wk old. Oral D-maltose gavage, everted sac, and Ussing chamber experiments were carried out to investigate intestinal glucose absorption and mRNA expression of glucose-transport-related genes as well as jejunal maltase activity. Upon gavage, glucose concentrations peaked at 10 min post-administration in 1-wk-old chicks, while they peaked at 40 min in 5-wk-old chickens. Glucose concentrations at 10 min were significantly higher in the 1-wk-old chicks (P = 0.010). Using the everted sacs experimental setup, 5 intestinal regions i.e., duodenum, proximal jejunum, distal jejunum, proximal ileum, and distal ileum, were targeted to examine D-maltose digestion and glucose transport across the intestinal mucosa. In the distal and proximal ileum, glucose concentrations were found to be significantly higher in the serosal compartment of the 1-wk-old chicks upon incubation with D-maltose (25 mM) (P < 0.05), while in the mucosal compartment the levels were significantly higher in the 5-wk-old chickens (P < 0.05). An Ussing chamber setup was employed to measure glucose-induced short-circuit current (ΔIsc) in the mucosal epithelium of the jejunum. In response to the addition of D-maltose (10 mM) into the mucosal compartment, ΔIsc was significantly higher in the 1-wk-old chicks (P = 0.018). Furthermore, no variations in jejunal maltase activity were observed between the 2 age groups. While jejunal glucose absorption was lower in the 5-wk-old chickens, the mRNA expression levels of jejunal SGLT1, GLUT2, and Na+/K+-ATPase did not show any significant differences between the 2 age groups. Our results suggest that the active transport of glucose across the jejunal epithelium decreases upon growth in broiler chickens but is not accompanied by any variations in maltase activity or in the expression of glucose-absorption-related genes.

Evidence for the role of INSL3 on sperm production in boars by passive immunisation.

Insulin-like factor 3 (INSL3), previously called relaxin-like factor, is essential for foetal testis descent and has been implicated in sperm production in adult males. This study investigated the role of INSL3 in sperm production by examining the effect of neutralising INSL3 by passive immunisation on testicular function and sperm output in boars. Six male Duroc boars were randomly assigned to passive immunisation and control groups (n = 3 each). The immunisation group was intravenously injected with an IgG fraction of anti-INSL3 antibody developed against the B domain of INSL3 at 2-week intervals from 21-40 weeks of age. The control group was treated with normal IgG in the same manner. Antibody administration reduced testis weight and caused a fourfold increase in the frequency of apoptotic germ cells, which was associated with upregulation of the pro-apoptotic caspase 3 and BAX, and downregulation of the anti-apoptotic XIAP and BCL2, and a substantial marked reduction in sperm concentration. Neutralising INSL3 delivered by passive immunisation reduced testis weight and sperm concentration by inducing germ cell apoptosis, suggesting that INSL3 acts as a germ cell survival/anti-apoptotic factor in the maintenance of sperm production.

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.

Prototyping the Experimental Setup to Quantify the Tissue Oxygen Consumption Rate and Its Preliminary Test.

In order to establish a reliable and practical method to make a diagnosis on the viability of an amputated extremity, we propose a method to evaluate the oxygen consumption rate. To validate this concept, we prototyped an experimental system with which the oxygen transfer rate into tissue can be assessed by the rate of change of the decrease in dissolved oxygen (DO) concentration within the buffer fluid surrounding the target tissue. The purpose of this study is to examine the feasibility of our prototyped experimental system by comparison between fresh and non-fresh rat skeletal muscles. The results show that the fresher tissue transferred more oxygen to the tissue, which suggests that tissue oxygen consumption is highly related to tissue freshness and can indirectly assess the tissue viability.

Intraoperative Oxygen Consumption During Liver Transplantation.

BACKGROUND: The aim of this study was to investigate the changes in oxygen consumption during liver transplantation and to examine the relationship between intraoperatively elevated systemic oxygen consumption and postoperative liver function. METHODS: This study was performed in 33 adult patients undergoing liver transplantation between September 2011 and March 2014. We measured intraoperative oxygen consumption through the use of indirect calorimetry, preoperative and intraoperative data, liver function tests, and postoperative complications and outcomes. RESULTS: The mean age of patients was 52 ± 9.7 years; 14 (42%) of them were women. Average Model for End-Stage Liver Disease scores were 20 ± 8.9. Oxygen consumption significantly increased after reperfusion from 172 ± 30 mL/min during the anhepatic phase to 209 ± 30 mL/min (P < .0001). We divided patients into 2 groups according to the increase in oxygen consumption after reperfusion (oxygen consumption after reperfusion minus anhepatic phase oxygen consumption: 40 mL/min increase as cutoff). The higher consumption group had a longer cold ischemia time and higher postoperative aspartate aminotransferase and alanine aminotransferase levels as compared with the lower oxygen consumption group. There were no statistically significant differences in major postoperative complications, but the higher oxygen consumption group tended to have shorter hospital stays than the lower consumption group (58 versus 95 days). CONCLUSIONS: We have demonstrated that oxygen consumption significantly increased after reperfusion. Furthermore, this increased oxygen consumption was associated with a longer cold ischemia time and shorter hospital stays.

The effect of IL-17 on the production of proinflammatory cytokines and matrix metalloproteinase-1 by human periodontal ligament fibroblasts.

OBJECTIVES: To investigate the effects of IL-17 on IL-6, IL-1ß, and matrix metalloproteinase (MMP-1) production, and to compare the MMP-1 production between the individual and combined effects of IL-1ß and IL-6 in human periodontal ligament fibroblasts (HPDLF). MATERIALS AND METHODS: Human periodontal ligament fibroblasts were cultured with IL-17 for 0.5, 1, 4, 24, 48, and 72 h, and were cultured with IL-1ß, IL-6/sIL-6R, or a combination of IL-1ß and IL-6/sIL-6R for 24 h. To measure the mRNA levels of IL-6, IL-1ß, and MMP-1, total RNA was extracted from the cultured HPDLF, and a real-time PCR analysis was performed. The protein levels of IL-6, IL-1ß, and MMP-1 in supernatants were measured using enzyme-linked immunosorbent assays (ELISAs). RESULTS: IL-17 significantly increased the expression of IL-6 and MMP-1 mRNA and protein, while IL-17 transiently increased the expression of IL-1ß mRNA. The combination of IL-1ß and IL-6/sIL-6R induced significantly higher levels of MMP-1 protein than IL-1ß alone. CONCLUSIONS: IL-17 upregulated the production of IL-6 and MMP-1 sequentially in HPDLF. IL-6/sIL-6R may enhance the effects of IL-1ß on MMP-1 production. The present results suggest that IL-17 induces MMP-1 production not only directly, but also indirectly by promoting IL-6 production, thus resulting in the degradation of collagens in the PDL.

First observation of a (1,0) mode frequency shift of an electron plasma at antiproton beam injection.

The frequency shift of the center-of-mass oscillation, known as the (1,0) mode, of a trapped electron plasma and, furthermore, its time evolution were observed during the cooling of an injected antiproton beam for the first time. Here, antiprotons mixed with the electrons did not follow faster electron oscillations but contributed to the modification of the effective potential. The time evolution of the plasma temperature, deduced from the frequency shift of the excited (3,0) mode, suggested that there was an abnormal energy deposition of the antiproton beam in the electron plasma before thermalization.

Ganglion cyst arising from the composite occipito-atlanto-axial joint cavity in a cat.

A four-year-old, female spayed Domestic Longhaired cat was referred for evaluation with a two month history of initial inability to jump progressing to ambulatory tetraparesis. Magnetic resonance imaging studies demonstrated a cystic lesion arising from the composite occipito-atlanto-axial joint cavity and extending to the region of the occipital bone and the axis. The lesion surrounded the spinal canal, causing moderate dorsal spinal cord compression at the atlanto-occipital joint. A dynamic myelographic study demonstrated attenuation of the dorsal contrast column at the atlanto-occipital joint when the cervical spine was positioned in extension. Partial excision of the cyst capsule by a ventral approach resulted in long-term (64 months) resolution of clinical signs. Histological evaluation was consistent with a ganglion cyst. An intra-spinal ganglion cyst arising from the composite occipito-atlanto-axial joint cavity may be considered as an uncommon differential diagnosis for cats with cervical myelopathy.

Body mass index delineates ALS from FTD: implications for metabolic health.

Weight loss and catabolic changes are increasingly recognized as factors that influence outcomes in patients with amyotrophic lateral sclerosis (ALS). An association between disease progression and low BMI has been reported in ALS; however, it remains unknown whether low BMI occurs across all forms of ALS and whether BMI changes with the development of cognitive impairment across the spectrum between ALS and frontotemporal dementia (FTD). One hundred and three ALS patients (56 limb predominant, 18 bulbar predominant, 13 ALS plus, 16 ALSFTD) were recruited and compared to 19 behavioral variant FTD (bvFTD) patients and a group of age-matched healthy controls. BMI was measured at the initial clinical visit. Patients were characterized as underweight, normal, overweight or obese, based on the current World Health Organization (WHO) guidelines. Limb and bulbar ALS patients had significantly lower BMI than ALS plus, ALSFTD, and bvFTD patient groups. When BMI was categorized using WHO guidelines the majority of the limb and bulbar ALS patients were either underweight or normal weight, whilst the majority of the ALS plus, ALSFTD and bvFTD patients were either overweight or obese. On follow-up BMI assessment the limb and bulbar groups tended to decline whilst ALS plus, ALSFTD and bvFTD groups remained stable or increased. BMI is significantly higher in ALS individuals with cognitive deficits. The present findings have prognostic implications for disease progression and may help delineate the metabolic profile across the ALSFTD spectrum.

Utility of glucagon stimulation test in type 1 diabetes after pancreas transplantation.

BACKGROUND: Despite recent progress of immunosuppressive therapy with newly developed agents, long-term pancreatic graft survival after pancreas transplantation still remains low. Therefore, precise assessment of ß-cell function after pancreas transplantation is necessary. METHODS: Pancreatic ß-cell secretory activity was measured by means of the peripheral plasma fasting serum C-peptide (CPR) response to 1 mg of glucagon intravenously in 23 patients after pancreas transplantation. The utility of ΔCPR after injection was compared with other indices that reflect insulin secretion. RESULTS: When we performed the test, 6 patients still needed insulin injection after the transplantation. Mean CPR before and after glucagon intravenously were 1.9 ± 0.98 ng/mL and 4.6 ± 2.29 ng/mL, respectively. Fasting serum CPR, secretory unit of islet in transplantation (SUIT) index, and ΔCPR after glucagon injection were significantly different between insulin users and nonusers. During follow-up (501 ± 228 days), 3 patients could stop using insulin, and their increase of CPR (1.8 ± 0.5 ng/mL) was significantly higher than that in continuous insulin users (0.3 ± 0.3 ng/mL). CONCLUSION: Fasting CPR, SUIT index, and ΔCPR after glucagon injection could reflect ß-cell function for post-pancreas transplant patients, and glucagon stimulation test could give us additional information to predict insulin-free treatment.

Acute type-A aortic dissection with patent false lumen through to the abdominal aorta: effects of a conventional elephant trunk on malperfusion syndromes and narrowed true lumen.

BACKGROUND: Narrowed true lumen and patent false lumen through to the terminal aorta is a high-risk condition for malperfusion syndromes (MS) in acute type-A aortic dissection. It is important to ascertain how the true and false lumens behave after surgery. PATIENTS AND METHODS: We retrospectively investigated 45 patients with this pathology. The true lumen sizes at the narrowest levels above and below the superior mesenteric artery were followed by computed tomography after surgery (0-36 months). RESULTS: Thirty-seven MS were seen in 23 patients. Hospital mortality was 8.9%. The narrowed true lumen was not enlarged in the first 6 months with a patent false lumen. The elephant trunk procedure did not improve the true lumen size. An extremely narrowed (≤3 mm) true lumen was associated with a significantly high incidence of MS and mortality. CONCLUSIONS: High incidences of MS were observed in this particular pathology. An extremely narrowed true lumen was accompanied by a high incidence of MS and mortality.

Deep venous thrombosis among disaster shelter inhabitants following the March 2011 earthquake and tsunami in Japan: a descriptive study.

OBJECTIVES: A retrospective analysis of data collected during subject screening following Japan's March 2011 earthquake and tsunami was performed. We aimed to determine the incidence of deep venous thrombosis (DVT) among screened subjects and to identify risk factors associated with the development of DVT as independent variables. METHODS: Calf ultrasonography was undertaken in 269 subjects living in 21 shelters in Miyagi prefecture during the one-month period immediately following the March 2011 disaster. Information regarding the health and risk factors of subjects was collected by questionnaire and assessment of physical signs. RESULTS: Of the 269 evacuees screened, 65 (24%) met the criteria for calf DVT. We found lower limb trauma, reduced frequency of urination and sleeping in a vehicle to be independent positive predictors of DVT. CONCLUSIONS: Evacuees had an increased risk of developing DVT, associated with tsunami-related lower limb injury, immobility and dehydration.

What is wrong with meta-analysis? The importance of clinical heterogeneity in myocardial regeneration research.

The author discusses the significance and potential pitfalls in performing a meta-analysis underscoring the importance of a usual forgotten issue in meta-analysis called clinical heterogeneity. Clinical heterogeneity can mislead results and misinform clinicians. Practical examples from the literature are given, and the results of meta-analyses are compared with the results of subsequent large randomised clinical trials addressing similar questions from a historical and contemporary point of view, highlighting clinical heterogeneity. The contemporary aspect culminates with the presentation of a meta-analysis evaluating myocardial cell regeneration with an emphasis in clinical heterogeneity, helping clinicians to understand the issue and better appraise future meta-analyses.

Hemilaminectomy and vertebral stabilization for thoracolumbar intervertebral disc associated dynamic compression in 11 dogs.

OBJECTIVE: To describe the diagnostic findings, surgical technique and outcome in dogs with thoracolumbar intervertebral disc-associated dynamic compression. STUDY DESIGN: Retrospective case series. ANIMALS: Client owned dogs (n = 11). METHODS: Medical records (2005-2010) of dogs with a stress myelographic diagnosis of spinal cord injury due to thoracolumbar intervertebral disc-associated dynamic compression with inconclusive compression in the neutral myelographic views that had hemilaminectomy and vertebral stabilization were reviewed. Data on pre- and postoperative neurologic status, diagnostic findings, surgical techniques and outcomes were retrieved. Follow-up clinical and radiographic evaluations were performed immediately, and at approximately one, two, and six months postoperatively as well as at annual follow-up examinations. RESULTS: The stress myelography demonstrated distinct ventral dynamic compression due to bulging of the disc and additional dorsal compression due to infolding of the ligamentum flavum in some cases. The median percentage of post-stress reduction in spinal cord height on the lateral view was 18.0% (9.8-27.2%). All dogs recovered after surgery and at follow-up examinations were still ambulatory (median: 45 months, range: 7 to 94 months). CONCLUSIONS AND CLINICAL RELEVANCE: Thoracolumbar intervertebral disc degeneration may result in disc-associated dynamic compression. Stress myelography was an effective means of diagnosing this condition and hemilaminectomy with vertebral stabilization was an effective treatment resulting in long-term neurological improvement in all dogs.

Expression of epidermal growth factors and a tight junction protein in the nasal mucosa of patients with chronic hypertrophic rhinitis

Background: ErbB family receptors and tight junction proteins participate in the pathologic process including tissue remodelling of inflammatory diseases in the upper and lower respiratory tracts. This study aimed at investigating the expressions of erbB1, 2, 3, 4, and a tight junction protein, claudin-1, in the nasal mucosa of patients with chronic hypertrophic rhinitis. Methods: Inferior turbinates were collected from 10 turbinectomised patients with allergic and non-allergic chronic hypertrophic rhinitis. The expressions of erbB1, 2, 3, 4, and claudin-1 were examined by fluorescence immunohistochemistry and by quantitative real-time transcription-polymerase chain reaction (qRT-PCR). Results: All erbB1-4 and claudin-1 were detected, and mainly localised in the epithelial cells and nasal gland cells. The immunoreactivity for claudin-1 was positively correlated with the expressions of erbB1, 2 and 4, but negatively correlated with that of erbB3. The mRNA expressions of erbB1, 2 and 4 were positively correlated with one another, whereas the expression of erbB3 showed negative correlation with the immunoreactivity for erbB2 and 4. Conclusions: These results suggest a possible participation of erbBs and claudin-1 in tissue remodelling in chronic hypertrophic rhinitis (AU)

Differential cellular localization of antioxidant enzymes in the trigeminal ganglion.

Because of its high oxygen demands, neural tissue is predisposed to oxidative stress. Here, our aim was to clarify the cellular localization of antioxidant enzymes in the trigeminal ganglion. We found that the transcriptional factor Sox10 is localized exclusively in satellite glial cells (SGCs) in the adult trigeminal ganglion. The use of transgenic mice that express the fluorescent protein Venus under the Sox10 promoter enabled us to distinguish between neurons and SGCs. Although both superoxide dismutases 1 and 2 were present in the neurons, only superoxide dismutase 1 was identified in SGCs. The enzymes relevant to hydrogen peroxide degradation displayed differential cellular localization, such that neurons were endowed with glutathione peroxidase 1 and thioredoxin 2, and catalase and thioredoxin 2 were present in SGCs. Our immunohistochemical finding showed that only SGCs were labeled by the oxidative damage marker 8-hydroxy-2'-deoxyguanosine, which indicates that the antioxidant systems of SGCs were less potent. The transient receptor potential vanilloid subfamily member 1 (TRPV1), the capsaicin receptor, is implicated in inflammatory hyperalgesia, and we demonstrated that topical capsaicin application causes short-lasting mechanical hyperalgesia in the face. Our cell-based assay revealed that TRPV1 agonist stimulation in the presence of TRPV1 overexpression caused reactive oxygen species-mediated caspase-3 activation. Moreover, capsaicin induced the cellular demise of primary TRPV1-positive trigeminal ganglion neurons in a dose-dependent manner, and this effect was inhibited by a free radical scavenger and a pancaspase inhibitor. This study delineates the localization of antioxidative stress-related enzymes in the trigeminal ganglion and reveals the importance of the pivotal role of reactive oxygen species in the TRPV1-mediated caspase-dependent cell death of trigeminal ganglion neurons. Therapeutic measures for antioxidative stress should be taken to prevent damage to trigeminal primary sensory neurons in inflammatory pain disorders.

Effects of a liquid diet on the response properties of temporomandibular joint nociceptive neurons in the trigeminal subnucleus caudalis of growing rats.

OBJECTIVE: To investigate whether low mechanical loading on the temporomandibular joint (TMJ) when ingesting a liquid diet affects the response properties of neurons in the trigeminal spinal tract subnucleus caudalis (Sp5C) in growing rats. MATERIALS AND METHODS: Shortly after weaning, 2-week-old male rats were fed chow pellets (control) or a liquid diet (experimental). Firing activities of single sensory units were recorded from the Sp5C at 4, 5, 7, and 9 weeks. Neurons were functionally classified by their responsiveness to TMJ stimuli. The responses of Class II and III neurons to TMJ stimuli were investigated. RESULTS: In both neuron classes, the firing threshold in the experimental group was significantly lower than in the control group at all time points, but remained static in the control group throughout the experimental period, whereas it peaked in the experimental group at 4 weeks, decreased at 5 weeks, and remained stable thereafter until 9 weeks. Similarly, the initial firing frequency was significantly higher in the experimental group than in the control group, but remained static in the control group throughout the experimental period, whereas in the experimental group, it was at its lowest at 4 weeks, increased at 5 weeks, and stayed stable thereafter until 9 weeks. CONCLUSION: Differences in TMJ loading arising from variable diet consistency during growth may affect the functional characteristics of Sp5C neurons.