Genomics to accelerate genetic improvement in tilapia.

Selective breeding of tilapia populations started in the early 1990s and over the past three decades tilapia has become one of the most important farmed freshwater species, being produced in more than 125 countries around the globe. Although genome assemblies have been available since 2011, most of the tilapia industry still depends on classical selection techniques using mass spawning or pedigree information to select for growth traits with reported genetic gains of up to 20% per generation. The involvement of international breeding companies and research institutions has resulted in the rapid development and application of genomic resources in the last few years. GWAS and genomic selection are expected to contribute to uncovering the genetic variants involved in economically relevant traits and increasing the genetic gain in selective breeding programs, respectively. Developments over the next few years will probably focus on achieving a deep understanding of genetic architecture of complex traits, as well as accelerating genetic progress in the selection for growth-, quality- and robustness-related traits. Novel phenotyping technologies (i.e. phenomics), lower-cost whole-genome sequencing approaches, functional genomics and gene editing tools will be crucial in future developments for the improvement of tilapia aquaculture.

Validation of a Simplified SRS-Schwab Classification Using a Sagittal Modifier.

STUDY DESIGN: Multicenter, prospective study of consecutive adult spinal deformity (ASD) patients. OBJECTIVE: To Validate Schwab's classification accuracy for surgical indication, and to evaluate a simplified sagittal modifier. SUMMARY OF BACKGROUND DATA: The SRS-Schwab Radiologic Classification based on clinical impact parameters, offers 27 different sagittal classification possibilities regarding sagittal vertical alignment (SVA), pelvic tilt (PT), and pelvic incidence-lumbar lordosis (PI-LL). The high number of classification possibilities makes it complex to use. METHODS: Inclusion criteria were ASD patients, presenting at least 1 criteria: Cobb ≥ 20°, SVA ≥ 5 cm, thoracic kyphosis ≥ 60°, or PT ≥ 25°. A total of 1,004 patients (410 nonoperative and 594 operative) were classified regarding SVA, PT, and PI-LL (0, +, ++), and 27 possibilities were identified. Categories were formed by adding the number of + signs, considering PT, SVA, and PI-LL. Three specific categories were identified: Aligned: 0 +; Moderate deformity: 1 to 3+; and Severe deformity: 4 to 6+. A χ-square test was performed for surgical indication (operated or not) and an analysis of variance was performed to evaluate the relationship between categories and Oswestry Disability Index (ODI). Probability <.05 was considered significant. RESULTS: Significant differences for HRQoL scores and surgical indication were found in the 27 sagittal parameter possibilities. For nonoperative patients, 230 (56.1%) were classified as aligned, 145 (35.4%) as moderate, and 35 (8.5%) as severe. For operative patients, there were 200 (33.7%), 215 (36.2%), and 179 (30.1%) in each respective subgroup. For HRQoL scores and surgical indication, no significant differences were found within each category, but significant differences were found when comparing the subgroups. CONCLUSIONS: Despite the correlation between SRS-Schwab classification and surgical indication, it is complex to use, with a total of 27 possibilities regarding sagittal modifiers. This simplification into three categories offers more readability, without losing any significant information, and could replace Schwab sagittal modifiers. In association with other parameters, they could be used for decision-making. LEVEL OF EVIDENCE: Level II.

Non-destructive elemental analysis of a carbonaceous chondrite with direct current Muon beam at MuSIC.

Electron- or X-ray-induced characteristic X-ray analysis has been widely used to determine chemical compositions of materials in vast research fields. In recent years, analysis of characteristic X-rays from muonic atoms, in which a muon is captured, has attracted attention because both a muon beam and a muon-induced characteristic X-ray have high transmission abilities. Here we report the first non-destructive elemental analysis of a carbonaceous chondrite using one of the world-leading intense direct current muon beam source (MuSIC; MUon Science Innovative Channel). We successfully detected characteristic muonic X-rays of Mg, Si, Fe, O, S and C from Jbilet Winselwan CM chondrite, of which carbon content is about 2 wt%, and the obtained elemental abundance pattern was consistent with that of CM chondrites. Because of its high sensitivity to carbon, non-destructive elemental analysis with a muon beam can be a novel powerful tool to characterize future retuned samples from carbonaceous asteroids.

Bayesian genome-wide association analysis for body weight in farmed Atlantic salmon (Salmo salar L.).

We performed a genome-wide association study to detect markers associated with growth traits in Atlantic salmon. The analyzed traits included body weight at tagging (BWT) and body weight at 25 months (BW25M). Genotypes of 4662 animals were imputed from the 50K SNP chip to the 200K SNP chip using fimpute software. The markers were simultaneously modeled using Bayes C to identify genomic regions associated with the traits. We identified windows explaining a maximum of 3.71% and 3.61% of the genetic variance for BWT and BW25M respectively. We found potential candidate genes located within the top ten 1-Mb windows for BWT and BW25M. For instance, the vitronectin (VTN) gene, which has been previously reported to be associated with cell growth, was found within one of the top ten 1-Mb windows for BWT. In addition, the WNT1-inducible-signaling pathway protein 3, melanocortin 2 receptor accessory protein 2, myosin light chain kinase, transforming growth factor beta receptor type 3 and myosin light chain 1 genes, which have been reported to be associated with skeletal growth in humans, growth stimulation during the larval stage in zebrafish, body weight in pigs, feed conversion in chickens and growth rate of sheep skeletal muscle respectively, were found within some of the top ten 1-Mb windows for BW25M. These results indicate that growth traits are most likely controlled by many variants with relatively small effects in Atlantic salmon. The genomic regions associated with the traits studied here may provide further insight into the functional regions underlying growth traits in this species.

Isolation and characterization of halophilic lactic acid bacteria acting as a starter culture for sauce fermentation of the red alga Nori (Porphyra yezoensis).

AIMS: A screening test was conducted for environmental samples to isolate halophilic lactic acid bacteria (HLAB) that can act as a starter in a Nori (Porphyra yezoensis)-sauce culture. METHOD AND RESULTS: After 9 months of incubation of enrichment cultures added with 25 kinds of environmental samples, growth of HLAB-like microorganisms was observed in six cultures salted at a 15% w/w level, including culture samples originally from mesopelagic water taken from 321 m-depth and from mountain snow taken at 2450 m-height. Ten strains were isolated and characterized as Tetragenococcus halophilus based on sequence analysis of the 16S rRNA gene. The isolates were inoculated into a newly prepared Nori-sauce culture and were confirmed to be able to act as a starter culture while three reference strains of T. halophilus obtained from a culture collection could not grow in the same culture. CONCLUSIONS: Halophilic lactic acid bacteria strains that can make growth in a highly salted Nori-sauce culture were isolated from environmental samples for the first time. All the isolates were identified as T. halophilus. SIGNIFICANCE AND IMPACT OF THE STUDY: The isolated strains are expected to be utilized as a starter culture for manufacturing fermented seaweed-sauce, which will be the first fermented food products obtained from algae.

TNF-α/TNFR1 signaling promotes gastric tumorigenesis through induction of Noxo1 and Gna14 in tumor cells.

Helicobacter pylori infection induces chronic inflammation that contributes to gastric tumorigenesis. Tumor necrosis factor (TNF-α) is a proinflammatory cytokine, and polymorphism in the TNF-α gene increases the risk of gastric cancer. We herein investigated the role of TNF-α in gastric tumorigenesis using Gan mouse model, which recapitulates human gastric cancer development. We crossed Gan mice with TNF-α (Tnf) or TNF-α receptor TNFR1 (Tnfrsf1a) knockout mice to generate Tnf-/- Gan and Tnfrsf1a-/- Gan mice, respectively, and examined their tumor phenotypes. Notably, both Tnf-/- Gan mice and Tnfrsf1a-/- Gan mice showed similar, significant suppression of gastric tumor growth compared with control Tnf+/+ or Tnfrsf1a+/+ Gan mice. These results indicate that TNF-α signaling through TNFR1 is important for gastric tumor development. Bone marrow (BM) transplantation experiments showed that TNF-α expressed by BM-derived cells (BMDCs) stimulates the TNFR1 on BMDCs by an autocrine or paracrine manner, which is important for gastric tumor promotion. Moreover, the microarray analysis and colony formation assay indicated that NADPH oxidase organizer 1 (Noxo1) and Gna14 are induced in tumor epithelial cells in a TNF-α-dependent manner, and have an important role in tumorigenicity and tumor-initiating cell property of gastric cancer cells. Accordingly, it is possible that the activation of TNF-α/TNFR1 signaling in the tumor microenvironment promotes gastric tumor development through induction of Noxo1 and Gna14, which contribute to maintaining the tumor cells in an undifferentiated state. The present results indicate that targeting the TNF-α/TNFR1 pathway may be an effective preventive or therapeutic strategy for gastric cancer.

Resposta à seleção de características de desempenho e morfométricas de tilápia-do-nilo ao longo do período de cultivo / Selection response for performance and morphometrics traits of Nile tilapia along the culture period

Estudou-se a resposta à seleção para características de desempenho e morfométricas de tilápia-do-nilo (Oreochromis niloticus) em diferentes períodos de cultivo. Realizaram-se análises unicaracter para todas as características estudadas e as análises bicaracter, associando-se o ganho de peso diário, mensurado em quatro biometrias, com as características morfométricas obtidas ao final do período de cultivo. As herdabilidades estimadas para peso vivo, altura e comprimento de cabeça foram mais altas em análises unicaracter do que as estimadas em análises bicaracter. Os valores de correlações genéticas, fenotípicas, Pearson e Spearman foram positivas e crescentes com o avanço da idade dos animais. As respostas à seleção indireta foram 87% mais altas que as obtidas pela resposta direta, indicando que a seleção pode ser realizada precocemente, utilizando como critério de seleção o ganho de peso diário total medido em biometrias intermediárias, sem apresentar perdas expressivas no ganho genético.

The objective of this work was to study the selection response for performance and morphometrics traits in Nile tilapias (Oreochromis niloticus) in different measurements of farming. The one-trait analysis was performed for all traits studied and the two-traits analysis, were associated with the daily total weight gain in four measurement among performance and morphometrics traits in the fourth measurement. The heritabilities estimated for live weight, height and head length were higher in one-trait analysis than two-traits analysis. The genetic correlations, phenotypic, Pearson and Spearman among weight gain and all other measurements were higher along the farming. The indirect response was estimated at over 87% of direct response, which indicates that the selection may be performed early, using the daily total weight gain as selection criteria in intermediate measurement, without significant losses in genetic gain and contributing to reduction of cost production.

The maturation of grafted bone after posterior lumbar interbody fusion with an interbody carbon cage: a prospective five-year study.

We evaluated the maturation of grafted bone in cases of successful fusion after a one- or two-level posterior lumbar interbody fusion (PLIF) using interbody carbon cages. We carried out a five-year prospective longitudinal radiological evaluation of patients using plain radiographs and CT scans. One year after surgery, 117 patients with an early successful fusion were selected for inclusion in the study. Radiological evaluation of interbody bone fusion was graded on a 4-point scale. The mean grades of all radiological and CT assessments increased in the five years after surgery, and differences compared to the previous time interval were statistically significant for three or four years after surgery. Because the grafted bone continues to mature for three years after surgery, the success of a fusion should not be assessed until at least three years have elapsed. There were no significant differences in the longitudinal patterns of grafted bone maturity between iliac bone and local bone. However, iliac bone grafting may remodel faster than local bone.

Superior mesenteric vein thrombosis presented transient false positivity for lupus anticoagulant under heparin treatment.

A 24-year-old Japanese man was admitted because of massive haematemesis and melaena with persistent abdominal pain. Markedly bloody ascites and severely oedematous small intestine were recognized, and angiography then revealed superior mesenteric vein thrombosis. After resection of the necrotic small intestine, continuous intravenous infusion of heparin and urokinase was performed. This patient had no familial or personal history of thrombosis. On the 15th day after operation, an initial search for lupus anticoagulant revealed that the prothrombin time (PT) ratio and dilute activated partial thromboplastin time (aPTT) were positive under heparin treatment, without evidence of rheumatic or connective tissue disease. Thrombocytopenia was observed with a nearly normocellular bone marrow. A follow-up examination 1 year later still revealed an increased aPTT. However, all tests for antiphospholipid antibodies had been negative including dilute aPTT for about 2 years since the 15th day after operation. These findings suggest that, in this patient, superior mesenteric vein thrombosis has not been associated with primary antiphospholipid syndrome but is probably idiopathic. Positive tests for lupus anticoagulant in the initial period may be unreliable due to heparin treatment.

Plasma procalcitonin in sepsis and organ failure.

INTRODUCTION: Because the use of procalcitonin (PCT) as a marker of bacterial infection has been advocated, this study was carried out to determine the usefulness of plasma PCT in the early diagnosis and differentiation of patients with non-infectious systemic inflammatory response syndrome (SIRS) from those with sepsis, and the relationship between plasma PCT level and severity of organ failure. MATERIALS AND METHODS: Thirty-five patients with non-septic SIRS (n = 16), sepsis (n = 7) or septic shock (n = 12) were included in this study. PCT and C-reactive protein (CRP) levels were measured and sepsis-related organ failure assessment (SOFA) score was calculated for these patients. Plasma PCT was measured by immunoluminometric assay. RESULTS: The median (minimum, maximum) plasma PCT levels were 0.6 (0.1, 3.4) ng/mL in non-septic SIRS, 5.4 (0.9, 47.7) ng/mL in sepsis and 73.4 (9.6, 824.1) ng/mL in septic shock, and significant differences existed in plasma PCT levels among the three groups. The median (minimum, maximum) CRP levels were 13.8 (0.3, 48.8) mg/dL in non-septic SIRS, 23.3 (1.4, 26.6) mg/dL in sepsis and 17.4 (2.2, 34.1) mg/dL in septic shock, without significant differences among the three groups. A good correlation was found between plasma PCT level and SOFA score (rs = 0.766, P < 0.0001), although no correlation was found between CRP level and SOFA score. CONCLUSIONS: CRP is increased by inflammatory disease as well as infection and is therefore not a good indicator of infection in patients with severe SIRS. On the other hand, PCT is a good indicator of severity of sepsis and organ failure in patients with severe SIRS since PCT levels correlated with sepsis and SOFA scores. PCT level is useful for diagnosis of sepsis and as an indicator of severity of organ failure in patients with SIRS.

Genomic organization and mapping of mouse CDV (carnitine deficiency-associated gene expressed in ventricle)-1 and its related CDV-1R gene.

We have previously reported that CDV (carnitine deficiency-associated gene expressed in ventricle)-1 was a downregulated gene in the hypertrophied ventricle of carnitine-deficient juvenile visceral steatosis mice and that the related gene (CDV-1R) showed no tissue specificity and no sensitivity to carnitine deficiency. In the present paper, the CDV-1/1R gene was isolated from a mouse genomic BAC library, and the genomic structure was characterized. We found that the CDV-1/1R gene consisted of at least 19 exons and encompassed approximately 48 kb. The splice sites conformed to the GT-AG rule, and the CDV-1R mRNA containing 19 exons was processed. CDV-1 mRNA containing 5 exons was constructed from the 3' half of CDV-1R. The first exon of CDV-1 consisted of the 3' side (116 bp) of intron 14 and exon 15 (87 bp) of CDV-1R. The presumed promoter sequence for CDV-1 located in the intron 14 of CDV-1R contained the common TATA box and consensus binding sites for various transcription factors (Nkx-2.5, Spl, C/EBP, SRF, YY1, and CREB), which seem to play roles in the heart-specific expression and carnitine deficiency-associated suppression of CDV-1. In the upstream region of the CDV-1 promoter, we found two VNTRs, 13 repeats of GATA1, and 16 copies of STRE involved in yeast stress response. The CDV-1/1R gene was located close to DSMIT68 on mouse Chromosome (Chr) 5, corresponding to human Chr 12q24. All these data revealed that two mRNA species, CDV-1 and CDV-1R, are expressed tissue-specifically by using promoters peculiar to each transcript in a single gene.

The signaling pathway of cardiotrophin-1 is not activated in hypertrophied ventricles of carnitine-deficient juvenile visceral steatosis (JVS) mice.

Cardiotrophin-1 (CT-1) is a novel cytokine which is involved in the growth and survival of cardiac cells. We examined whether CT-1 plays a role in the development of cardiac hypertrophy in carnitine-deficient juvenile visceral steatosis (JVS) mice. The CT-1 mRNA level was quantitatively measured by the competitive RT-PCR method. In contrast to other models including spontaneously hypertensive rats, CT-1 mRNA in the ventricles of JVS mice was comparable to the control at 5 days and was less than half the control value at 2 and 8 weeks when the ventricles of the JVS mice were highly hypertrophied. There were no significant differences in CT-1 mRNA levels in the lung, liver, kidney, small intestine and skeletal muscle between the JVS and control mice at 2 weeks. We did not find any difference between JVS and control mice at 2 weeks in the mRNA level of ventricular leukemia inhibitory factor (LIF) which binds to the same receptor as CT-1. Furthermore, almost no phosphorylated STAT3 (signal transducer and activator of transcription 3), downstream of the LIF receptor and the gp130 signaling subunit, was observed in the ventricles of JVS and control mice. These data show that the CT-1 signaling pathway does not play a significant role in the development of cardiac hypertrophy in JVS mice. Furthermore, we could not detect any differences in insulin-like growth factor I and II mRNA levels. All these data suggest distinct differences in the mechanisms of cardiac hypertrophy between JVS mice and other model animals.

Expression of nm23 antimetastatic gene product in head and neck squamous cell carcinoma.

The nm23 gene has been implicated as a suppressor gene involved in the control of the metastatic process of malignant cells. Reduced levels of nm23 gene product have been found in tumor cells with high metastatic potential such as several types of rodent tumors and human breast, colorectal, and lung carcinoma. This pilot study examines the expression of the nm23 gene product compared with nodal status in 70 consecutive patients with squamous cell carcinomas of the head and neck. Immunohistochemical staining was carried out on these tumor tissues with a monoclonal antibody to nm23-H1 peptide sequence. The tissues were scored from 0 to 2 by 2 independent observers. Reduced immunoreactivity, grades 0 and 1, was observed in 30 patients with positive nodal status (N = 34, 88%). Strong immunoreactivity, grade 2, was observed in 20 patients with negative nodal status (N = 36, 56%). Reduced expression of nm23 gene product was observed in patients with positive lymph node metastasis (P = 0. 0006, chi(2)). However, no significant differences in survival of these groups based on nm23 expression could be shown with the Kaplan-Meier analysis. This initial finding in the difference of nm23 gene product expression in patients with differing lymph node status is exciting but must be further validated with future studies.

Deformation and vascular occlusion of the growing rat femoral head induced by mechanical stress.

To investigate the effects of mechanical stress on the extent of deformation and decrease of the blood supply to the femoral head in the growing rat, we conducted a histological and microangiographic study, using a custom-made hip-joint loading device. Application of a 1-kg to 3-kg load to the hip joint caused compression not only of the articular cartilage but also of the growth plate of the femoral head. These changes were particularly prominent in the lateral portion of the femoral head. The compression also caused complete occlusion of the lateral epiphyseal arteries, representing the blood supply to the femoral head, at the point of penetration into the epiphyseal cartilage. Our results indicate that the sum of forces during mechanical load application to the hip joint in growing rats acts to deform the cartilage of the lateral side of the femoral head. Our results also indicate the presence of a mechanically weak point in the femoral head which may lead to occlusion of the lateral epiphyseal arteries.

[Drug delivery system for the treatment of osteoporosis].

The number of agents for the treatment of osteoporosis has increased over the past few years allowing more and more differentiated therapy. Calcitonin, vitamin D3, bisphosphonate, estrogen, vitamin K2 and calcium are administered for the treatment of osteoporosis in Japan. The purpose of drug delivery system (DDS) is to provide a practical approach to increasing efficacy and minimizing side effects of the drugs. In fact by the development of DDS, topical administration, prodrug and targeting therapy are used in the treatment for osteoporosis. It is well known that calcitonin in susceptible to proteolysis in the intestine. So, calcitonin must be given intramuscular or subcutaneous injection for therapy. DDS of calcitonins is studying in many routes of administration, such as nasal, transdermal, ocular, oral, bronchial, rectal and vaginal.

Evaluation of continuous intraspinal narcotic analgesia for chronic pain from benign causes.

Intraspinal narcotic analgesia (INA) has been used for chronic pain from nonmalignant causes with moderate success. To ascertain the efficacy of the morphine pump, we reviewed the 2-year results of continuous INA in 18 patients with failed back syndrome or arachnoiditis and intractable, debilitating pain that was unrelieved by conventional means. All patients underwent a trial screening of single-dose intrathecal narcotics with good pain relief. After 2 years, 8 pumps were still functioning, 8 patients had the pump removed or turned off, and 2 patients were lost to follow-up. Our patients averaged 1.4 additional procedures or hospitalizations after initial pump insertion. Overall, only 4 patients had objective evidence of benefit from INA, for a success rate of 25%. Results of this review suggest INA should not be used for the long-term management of chronic pain from nonmalignant causes.

Characteristics of blood vessels feeding the femoral head liable to osteonecrosis in spontaneously hypertensive rats.

In order to elucidate the cause of osteonecrosis of the femoral head in spontaneously hypertensive rats (SHRs), which resembles the osteonecrosis of Perthes' disease, we observed the three-dimensional structure of vascular casts of the blood vessels feeding the femoral head using both optical and scanning electron microscopes. During the period of 9-15 weeks after birth, when osteonecrosis of the femoral heads in SHRs occurred frequently, the lateral epiphyseal vessels (LEVs), which were the main feeding vessels, entered from the lateral of the femoral heads. Anastomosing branches of LEVs between the epiphysis and the femoral neck were scarce even in the femoral heads showing normal ossification. It seemed that the development of LEVs in SHRs did not proceed normally in this period. Furthermore, remarkable segmental stenosis and the obstruction of LEVs were often recognized near the lateral of the femoral heads. These results suggest that LEVs in growing SHRs have the vascular structure that could cause an interruption of the blood supply to the femoral heads.

[Does a tracheal bifurcation shift upward due to increased intraabdominal pressure during laparoscopic surgery?].

To evaluate the influence of increased intraabdominal pressure on a position of tracheal bifurcation, we measured the distance from the tracheal bifurcation to the tip of the endotracheal tube which is fixed on a patient's mouth during a laparoscopic surgery. The distance decreased from 3.5 +/- 1.9 to 3.1 +/- 1.9 cm (P < 0.05) at mean intraabdominal pressure of 8.7 mmHg. This result suggests that the ventilation may become difficult due to the occlusion of the tip of the endotracheal tube by a tracheal bifurcation or due to inadvertent single lung ventilation under inappropriately deep intubation.

Degradation-resistant character of synthetic hydroxyapatite blocks filled in bone defects.

The long-term changes of both the implanted hydroxyapatite (HA) blocks and the bone around them were observed radiologically to investigate the clinical usefulness of HA. HA blocks were used as a space filler in surgically created bone defects of seven cases due to curettage of bone tumours or removal for bone grafts, and they were followed up for 78 to 109 months. Bone formation around HA blocks peaked within 1 year after implantation, and then it faded gradually. However, shapes of HA blocks were rarely changed in all the cases. A clear zone around HA blocks was never observed . HA blocks were biocompatible and demonstrated a character resistant to degradation.