Genome and Phylogenetic Analysis of Cucumber Green Mottle Mosaic Virus Global Isolates and Validation of a Highly Sensitive RT-qPCR Assay.

The last two decades have seen exponential growth in the international movement of seeds for annual food crops, from a gross U.S. import value of $349 million in 1999 to $1.05 billion in 2019. This has led to the proportionate growth of seedborne pathogens dispersed with seed stocks. One such viral pathogen is cucumber green mottle mosaic virus (CGMMV), a tobamovirus that infects cucurbit crops such as melon, watermelon, cucumber, pumpkin, and squash. The first CGMMV introduction to California occurred in 2013, with subsequent annual outbreaks or detections since then. Here, we describe the use of next-generation sequencing to characterize the full genomes of 25 CGMMV isolates collected between 2013 and 2020 in California, either from CGMMV field detections or seed lots identified as CGMMV positive. We sequenced an additional 31 CGMMV isolates collected in Europe, Israel, and southeast Asia that were provided by industry collaborators. We also performed an in silico nucleotide database search in GenBank for full genome CGMMV sequences to include in all in silico analyses. Based on conserved regions within the coat protein gene, we then developed a quantitative reverse-transcription PCR assay for the sensitive and specific detection of CGMMV in seed and plant samples. Finally, based on our sequence and phylogenetic analysis, our data support that CGMMV has been introduced multiple times into California.

Sclerostin and bone turnover markers response to cycling and running at the same moderate-to-vigorous exercise intensity in healthy men.

BACKGROUND: Recreational cycling is a popular activity which stimulates and improves cardiovascular fitness. The corresponding benefits for bone are unclear. PURPOSE: This study examined the effect of running (high-impact) vs. cycling (low-impact), at the same moderate-to-vigorous exercise intensity, on markers of bone formation (N-terminal propeptide of type I collagen, PINP) and bone resorption (C-telopeptide of type I collagen, CTX-1), a non-collagenous bone remodeling marker (osteocalcin), as well as bone-modulating factors, including parathyroid hormone (PTH), irisin (myokine) and sclerostin (osteokine). METHODS: Thirteen healthy men (23.7 ± 1.0 y) performed two progressive exercise tests to exhaustion (peak VO2) on a cycle ergometer (CE) and on a treadmill (TM). On subsequent separate days, in randomized order, participants performed 30-min continuous running or cycling at 70% heart rate reserve (HRR). Blood was drawn before, immediately post- and 1 h into recovery. RESULTS: PTH transiently increased (CE, 51.7%; TM, 50.6%) immediately after exercise in both exercise modes. Sclerostin levels increased following running only (27.7%). Irisin increased following both running and cycling. In both exercise modes, CTX-1 decreased immediately after exercise, with no significant change in PINP and osteocalcin. CONCLUSION: At the same moderate-to-vigorous exercise intensity, running appears to result in a greater transient sclerostin response compared with cycling, while the responses of bone markers, PTH and irisin are similar. The longer-term implications of this differential bone response need to be further examined.

Effects of High-Intensity Interval Running Versus Cycling on Sclerostin, and Markers of Bone Turnover and Oxidative Stress in Young Men.

This study compared sclerostin's response to impact versus no-impact high-intensity interval exercise in young men and examined the association between exercise-induced changes in sclerostin and markers of bone turnover and oxidative stress. Twenty healthy men (22.3 ± 2.3 years) performed two high-intensity interval exercise trials (crossover design); running on treadmill and cycling on cycle ergometer. Trials consisted of eight 1 min running or cycling intervals at ≥ 90% of maximal heart rate, separated by 1 min passive recovery intervals. Blood samples were collected at rest (pre-exercise), and 5 min, 1 h, 24 h, and 48 h following each trial. Serum levels of sclerostin, cross-linked telopeptide of type I collagen (CTXI), procollagen type I amino-terminal propeptide (PINP), thiobarbituric acid reactive substances (TBARS), and protein carbonyls (PC) were measured. There was no significant time or exercise mode effect for PINP and PC. A significant time effect was found for sclerostin, CTXI, and TBARS with no significant exercise mode effect and no significant time-by-mode interaction. Sclerostin increased from pre- to 5 min post-exercise (47%, p < 0.05) and returned to baseline within 1 h following the exercise. CTXI increased from pre- to 5 min post-exercise (28%, p < 0.05), then gradually returned to baseline by 48 h. TBARS did not increase significantly from pre- to 5 min post-exercise but significantly decreased from 5 min to 48 h post-exercise. There were no significant correlations between exercise-induced changes in sclerostin and any other marker. In young men, sclerostin's response to high-intensity interval exercise is independent of impact and is not related to changes in bone turnover and oxidative stress markers.

Response of Sclerostin and Bone Turnover Markers to High Intensity Interval Exercise in Young Women: Does Impact Matter?

This study examined potential exercise-induced changes in sclerostin and in bone turnover markers in young women following two modes of high intensity interval exercise that involve impact (running) or no-impact (cycling). Healthy, recreationally active, females (n=20; 22.5±2.7 years) performed two exercise trials in random order: high intensity interval running (HIIR) on a treadmill and high intensity interval cycling (HIIC) on a cycle ergometer. Trials consisted of eight 1 min running or cycling intervals at ≥90% of maximal heart rate, separated by 1 min passive recovery intervals. Blood samples were collected at rest (pre-exercise) and 5 min, 1h, 24h, and 48h following each exercise trial. Serum was analyzed for sclerostin, cross linked telopeptide of type I collagen (CTXI), and procollagen type I amino-terminal propeptide (PINP). A significant time effect was found for sclerostin, which increased from pre-exercise to 5 min after exercise in both trials (100.2 to 131.6 pg/ml in HIIR; 102.3 to 135.8 pg/ml in HIIC, p<0.001) and returned to baseline levels by 1h, with no difference between exercise modes and no exercise mode-by-time interaction. CTXI did not significantly change following either trial. PINP showed an overall time effect following HIIR, but none of the post hoc pairwise comparisons were statistically significant. In young women, a single bout of high intensity exercise induces an increase in serum sclerostin, irrespective of exercise mode (impact versus no-impact), but this response is not accompanied by a response in either bone formation or resorption markers.

Mycoviruses of an endophytic fungus can replicate in plant cells: evolutionary implications.

So far there is no record of a specific virus able to infect both fungal and plant hosts in nature. However, experimental evidence shows that some plant virus RdRPs are able to perform replication in trans of genomic or DI RNAs in the yeast Saccharomyces cerevisiae. Furthermore, tobacco mosaic virus was recently shown to replicate in a filamentous ascomycetous fungus. Thus, at least experimentally, some plant viruses can infect some fungi. Endophytic fungi have been reported from many plants and several of these fungi have been shown to contain viruses. Here we tested if mycoviruses derived from a marine plant endophyte can replicate in plant cells. For this purpose, we used partially purified viral particles from isolate MUT4330 of Penicillium aurantiogriseum var. viridicatum which harbors six virus species, some having dsRNA and some positive-strand ssRNA genomes. These were transfected into three distinct plant protoplast cell systems. Time-course analysis of absolute RNA accumulation provided for the first time evidence that viruses of two species belonging to the Partitiviridae and Totiviridae families, can replicate in plant cells without evidence of host adaptation, i.e, changes in their nucleotide sequence.

Identification of the cleavage sites of the RNA2-encoded polyproteins for two members of the genus Torradovirus by N-terminal sequencing of the virion capsid proteins.

Torradoviruses, family Secoviridae, are emergent bipartite RNA plant viruses. RNA1 is ca. 7kb and has one open reading frame (ORF) encoding for the protease, helicase and RNA-dependent RNA polymerase (RdRp). RNA2 is ca. 5kb and has two ORFs. RNA2-ORF1 encodes for a putative protein with unknown function(s). RNA2-ORF2 encodes for a putative movement protein and three capsid proteins. Little is known about the replication and polyprotein processing strategies of torradoviruses. Here, the cleavage sites in the RNA2-ORF2-encoded polyproteins of two torradoviruses, Tomato marchitez virus isolate M (ToMarV-M) and tomato chocolate spot virus, were determined by N-terminal sequencing, revealing that the amino acid (aa) at the -1 position of the cleavage sites is a glutamine. Multiple aa sequence comparison confirmed that this glutamine is conserved among other torradoviruses. Finally, site-directed mutagenesis of conserved aas in the ToMarV-M RdRp and protease prevented substantial accumulation of viral coat proteins or RNAs.

RNA1-Independent Replication and GFP Expression from Tomato marchitez virus Isolate M Cloned cDNA.

Tomato marchitez virus (ToMarV; synonymous with Tomato apex necrosis virus) is a positive-strand RNA virus in the genus Torradovirus within the family Secoviridae. ToMarV is an emergent whitefly-transmitted virus that causes important diseases in tomato (Solanum lycopersicum) in Mexico. Here, the genome sequence of the ToMarV isolate M (ToMarV-M) was determined. We engineered full-length cDNA clones of the ToMarV-M genomic RNA (RNA1 and RNA2), separately, into a binary vector. Coinfiltration of both triggered systemic infections in Nicotiana benthamiana, tomato, and tomatillo (Physalis philadelphica) plants and recapitulated the biological activity of the wild-type virus. The viral progeny generated from tomato and tomatillo plants were transmissible by the whitefly Bemisia tabaci biotype B. Also, we assessed whether these infectious clones could be used for screening tomato cultivars for resistance to ToMarV and our results allowed us to differentiate resistant and susceptible tomato lines. We demonstrated that RNA1 of ToMarV-M is required for the replication of RNA2, and it can replicate independently of RNA2. From this, ToMarV-M RNA2 was used to express the green fluorescent protein in N. benthamiana plants, which allowed us to track cell-to-cell movement. The construction of full-length infectious cDNA clones of ToMarV-M provides an excellent tool to investigate virus-host-vector interactions and elucidate the functions of torradovirus-encoded proteins or the mechanisms of replication of torradovirus genomic RNA.

A new tobamovirus infecting tomato crops in Jordan.

In this study, we completed the whole genome sequence of a new tobamovirus isolated from tomato plants grown in greenhouses in Jordan during the spring of 2015. The 6393-nt single-stranded RNA (ssRNA) genome encodes four proteins, as do other tobamoviruses: two replication-related proteins of 126 kDa and 183 kDa, a 30-kDa movement protein (MP) and a 17.5-kDa coat protein (CP). Phylogenetic analysis showed that this virus does not group with either the tomato mosaic virus (ToMV) or the tobacco mosaic virus (TMV) clades. Instead, it stems from a branch leading to the TMV clade. Analysis of possible recombination events between this virus and representative isolates of closely related tomato-infecting tobamoviruses showed that at least one region originated by recombination. We provide evidence that we have identified a new tobamovirus, for which we propose the name "tomato brown rugose fruit virus".

Differential sclerostin and parathyroid hormone response to exercise in boys and men.

SUMMARY: Physical exercise benefits bone structure and mineralization, especially in children. Immediately following high-impact exercise, PTH increased and returned to resting values within 24 h in both groups, while sclerostin increased in men but not in boys. The underlying mechanisms and implication of this age-related differential response are unclear. INTRODUCTION: Circulating sclerostin, a negative regulator of bone, decreases during puberty and increases in adulthood. Parathyroid hormone (PTH) is inversely related to sclerostin. In mice, sclerostin decreases following 24 h of mechanical stimulation. Its response to exercise in humans and, especially in children, in whom high-impact physical exercise benefits bone structure and mineralization is unclear. The aim of this study was to investigate the acute response of sclerostin to a single exercise session of high mechanical loading and the corresponding changes in PTH in boys and men. METHODS: Twelve boys (10.2 ± 0.4 years old) and 17 young men (22.7 ± 0.8 years old) underwent a protocol of plyometric exercises (total 144 jumps). Blood samples were collected pre-, 5 min, 1 h, and 24 h post-exercise. RESULTS: Boys had significantly higher resting values of sclerostin compared with men (150 ± 37 vs. 111 ± 34 pg/ml, respectively, p = 0.006). Following exercise, sclerostin markedly increased in men but this response was attenuated in boys (at 5 min: 51 ± 38 vs. 14 ± 21%, respectively, p = 0.005). PTH levels were similar in boys and men at rest and throughout the 24-h study period, increasing significantly (p < 0.001) 5 min after exercise, decreasing after 60 min post-exercise and returning to resting values within 24 h. CONCLUSION: Although the PTH response was similar in boys and men, the sclerostin response was greater in men. The combined increases in PTH and sclerostin immediately post-exercise appear contrary to the accepted osteogenic effect of exercise. The underlying mechanisms and full implication of the differential response between children and adults need to be further examined.

Markers of biological stress and mucosal immunity during a week leading to competition in adolescent swimmers.

In this study we examined changes in the salivary concentrations of immunoglobulin A (sIgA), cortisol (sC), testosterone (sT), and testosterone-to-cortisol ratio (T/C) in 21 competitive swimmers, 11-15 years old, during a week leading to competition as compared to a control (noncompetition) week. No day-to-day changes or significant differences between weeks were observed for sIgA (47.9 ± 4.4 versus 54.9 ± 5.2 µg/mL for control versus competition week, resp.), sC (2.7 ± 0.2 versus 2.5 ± 0.2 ng/mL for control versus competition week, resp.), and T/C ratio (83.4 ± 7.0 versus 77.9 ± 7.7 for control versus competition week, resp.). In contrast, sT was significantly lower during the week of competition (154.5 ± 11.3 pg/mL) as compared to the control week (181.3 ± 11.5 pg/mL) suggesting that the swimmers were in a catabolic state, although this did not have a negative effect on their performance. In conclusion, salivary cortisol did not change between the two weeks, and thus competition stress was relatively low, and mucosal immunity was unaffected in these young athletes prior to competition.

Mapping of QTL for Tolerance to Cereal Yellow Dwarf Virus in Two-rowed Spring Barley.

Cereal yellow dwarf virus (CYDV-RPV) causes a serious viral disease affecting small grain crops around the world. In the United States, it frequently is present in California where it causes significant yield losses, and when infections start early in development, plant death. CYDV is transmitted by aphids, and it has been a major impediment to developing malting barley in California. To identify chromosome locations associated with tolerance/resistance to CYDV, a segregating population of 184 recombinant inbred lines (RIL) from a cross of the California adapted malting barley line Butta 12 with the CYDV tolerant Madre Selva was used to construct a genetic map including 180 polymorphic markers mapping to 163 unique loci. Tolerance to CYDV was evaluated in replicated experiments where plants were challenged by aphid mediated inoculation with the isolate CYDV-RPV in a controlled environment. Quantitative trait loci (QTL) analysis revealed the presence of two major QTL for CYDV tolerance from Madre Selva on chromosomes 2H (Qcyd.MaBu-1) and 7H (Qcyd.MaBu-2), and 4 minor QTL from Butta 12 on chromosomes 3H, 4H, and 2H. This paper discusses the contribution of each QTL and their potential value to improve barley tolerance to CYDV.

First Report of Cucumber green mottle mosaic virus on Melon in the United States.

In July 2013, a melon (Cucumis melo var. Saski) field in Yolo County, California, was inspected as part of a phytosanitary inspection for seed production. The leaves of the plants showed mosaic, green mottle, and blotches. When plant sap was examined using a transmission electron microscope, rigid rod-shaped particles were observed. Melon plant samples were analyzed by both CDFA and USDA APHIS PPQ laboratories and tested positive using DAS-ELISA against Cucumber green mottle mosaic virus (CGMMV) (Agdia, Elkhart, IN). To confirm the presence of CGMMV, total RNA was analyzed by RT-PCR using primers CGMMV-F5370 5'-CTAATTATTCTGTCGTGGCTGCGGATGC-3' and CGMMV-R6390 5'-CTTGCAGAATTACTGCCCATA-3' designed by PPQ based on 21 genomic sequences of CGMMV found worldwide. The 976-bp amplicon was sequenced (GenBank Accession No. KJ453559) and BLAST analysis showed the sequence was 95% identical to MP and CP region of CGMMV isolates reported from Russia (GQ495274, FJ848666), Spain (GQ411361), and Israel (KF155231), and 92% to the isolates from China (KC852074), Korea (AF417243), India (DQ767631), and Japan (D12505). These analyses confirm the virus was CGMMV. To our knowledge, this is the first report of CGMMV in the United States. Based on our sequence data, a second set of primers (CGMMV-F5796 5'-TTGCGTTTAGTGCTTCTTATGT-3' and CGMMV-R6237 5'-GAGGTGGTAGCCTCTGACCAGA-3'), which amplified a 440-bp amplicon from CGMMV CP region, was designed and used for testing all the subsequent field and seed samples. Thirty-seven out of 40 randomly collected Saski melon samples tested positive for CGMMV, suggesting the virus was widespread in the field. All the melon samples also tested positive for Squash mosaic virus (SqMV) using DAS-ELISA (Agdia). Therefore, the symptoms observed likely resulted from a mixed infection. The melon field affected by CGMMV was immediately adjacent to fields of cucumber (Cucumis sativus var. Marketmore 76) and watermelon (Citrullus lanatus var. Sugar Baby) crops, both for seed production with no barrier between the crops. CGMMV was also detected from symptomatic plants from both fields. Seed lots used for planting all three crops were tested and only the melon seed was positive for CGMMV, suggesting the seed as the source of infection. The sequenced 440-bp RT-PCR amplicons from CGMMV-infected cucumber and watermelon plants and melon seeds were 99% identical to the CGMMV from the field melon. A cucumber plant infected with CGMMV but not SqMV was used for mechanical inoculation at the Contained Research Facility at University of California, Davis. Inoculated cucumber, melon, and watermelon plants showed green mottle and mosaic similar to that observed in the field. CGMMV is a highly contagious virus and damage by this virus on cucurbit crops has been reported in regions where CGMMV is present (2). CGMMV was detected on cucumber grown in greenhouses in Canada with 10 to 15% yield losses reported due to this virus (1). The three cucurbit crops in Yolo County were planted in an isolated area with no other cucurbits nearby. Measures, including destroying all the cucurbit plant material, have been taken to eradicate the virus. Use of CGMMV free cucurbit seed is necessary for prevention of this disease. References: (1) K.-S. Ling et al. Plant Dis. 98:701, 2014. (2) J. Y. Yoon et al. J. Phytopathol. 156:408, 2008.

Do molecules matter more than morphology? Promises and pitfalls in parasites.

Systematics involves resolving both the taxonomy and phylogenetic placement of organisms. We review the advantages and disadvantages of the two kinds of information commonly used for such inferences--morphological and molecular data--as applied to the systematics of metazoan parasites generally, with special attention to the malaria parasites. The problems that potentially confound the use of morphology in parasites include challenges to consistent specimen preservation, plasticity of features depending on hosts or other environmental factors, and morphological convergence. Molecular characters such as DNA sequences present an alternative data source and are particularly useful when not all the parasite's life stages are present or when parasitaemia is low. Nonetheless, molecular data can bring challenges that include troublesome DNA isolation, paralogous gene copies, difficulty in developing molecular markers, and preferential amplification in mixed species infections. Given the differential benefits and shortcomings of both molecular and morphological characters, both should be implemented in parasite taxonomy and phylogenetics.

Reliability and validity of the lactate-minimum test. A revisit.

AIM: The Lactate-Minimum Test (LMT) is a high-resolution, physiologically elegant test for estimating the anaerobic threshold (AnT), or the Maximal Lactate Steady-State (MLSS). Nevertheless, it has not gained the acceptance level of typical progressive lactate-response tests (PLRT). Aim of this study was to compare LMT's validity and reviewer reliability vs. a PLRT-type test and re-evaluate the justification for LMT's dismissal. METHODS: Sixteen male distance trained runners (37.1±11.6 yrs) were included in the study. MLSS, LMT, and PLRT tests were performed in separate sessions. Two reviewers, blind to the subjects' identity, independently determined LMT and PLRT's threshold velocities (VLMT, VPLRT) twice. Additionally, VLMT was determined objectively, using best-fit polynomial regressions (VLMTP). RESULTS: VPLRT, VLMT and VLMTP correlated well with VMLSS (r=0.92, 0.90, 0.93, resp.). VPLRT was identical to VMLSS (13.54 km·h-1), but VLMT and VLMTP were 0.33 and 0.46 km·h-1 lower, respectively. Inter-reviewer reliability was higher for VLMT than VPLRT (ICC=0.96 vs. 0.57, resp.). Intra-reviewer reliability showed a similar pattern. CONCLUSION: LMT's underestimation of MLSS appears corrigible. The validity of corrected LMT appears comparable to that of PLRT, while its reliability, objectivity and resolution are superior. Although neither test is a perfect MLSS-substitute, the corrected LMT is not inferior to PLRT-type testing and cannot be dismissed.

Maturity status in male child and adolescent athletes.

AIM: Early-maturing individuals may be at an advantage in some sports. The purpose of this study was to compare maturity status between competitive male child (10-12 years old) and adolescent (14-16 years old) athletes and minimally-active, age-matched controls. METHODS: In total, 224 males were included in the study. Children (n=115) included minimally-active boys (n=34), soccer players (n=26), gymnasts (n=25) and hockey players (n=30). Adolescents (n=109) included minimally-active adolescents (n=31), soccer players (n=30), gymnasts (n=17) and hockey players (n=31). Sexual maturity was assessed using secondary sex characteristics and salivary testosterone concentration (sT). Skeletal age was also assessed, using quantitative ultrasound (Sunlight BonAgeTM). RESULTS: Within each age group, no differences were observed between sport groups in chronological age, sT or pubertal age. In children, hockey players were more skeletally mature (12.43±1.36 years) than all other groups (11.0±1.0; 11.6±1.4 and 11.7±1.4 years for soccer, gymnasts and controls, respectively). In adolescents, hockey players and gymnasts had higher skeletal maturity (16.8±1.5 and 16.9±1.6 years, respectively; P<0.05) than controls (15.99±1.13 years). CONCLUSION: While sexual and hormonal maturity does not appear to differ between similar-aged athletes of different sports, the results suggest greater skeletal maturity in hockey players, even before puberty.

Physical activity participation and bleeding characteristics in young patients with severe haemophilia.

Patients with haemophilia are now widely advised to participate in sport activities. However, no extensive data are available about their actual participation. The aim of this study was to describe the type; intensity and duration of leisure time physical activity (PA) among young patients with severe hemophilia and to assess whether there are differences in bleeding profile and muscle strength in related to activity intensity. Forty-four boys (ages 12-25 years) with severe haemophilia were studied. PA was assessed by the Godin and Shephard (G&S) questionnaire. Bleeding profile was determined based on a one month diary filled by each patient. Muscle strength of the lower limbs muscles was assessed using a hand held dynamometer. Only three subjects did not perform any PA. Twenty-five of the participants performed strenuous PA at least once a week. An inverse, moderate association (r(p) =-0.45, P < 0.002) was found between the G&S score and age. There were no significant differences in bleeding frequency or pain but a significant difference in the cause of bleed was found: those who exercised strenuously showed a higher proportion of bleeds due to traumatic reasons (P < 0.01). No differences in muscle strength values were noted in related to activity intensity also no linear association was noted between muscle strength and bleeding profile. Further investigation, including prospective studies, is needed in order to assess the temporal sequencing between training and the occurrence of bleeds and bleeds cause.

A New Tospovirus sp. in Cucurbit Crops in Mexico.

During the 2007 growing season, melon (Cucumis melo) samples from the state of Guerrero in Mexico showing mosaic and other virus-like symptoms were collected for analysis. Electron microscopic examination of negatively stained leaf-dip extracts revealed the presence of abundant virus-like particles with features characteristic of the family Bunyaviridae. No other viral particles were observed in these preparations. However, enzyme-linked immunosorbent assays (ELISAs) specific for the most common Tospovirus spp. gave negative results. Antibodies raised against purified nucleocapsids reacted specifically with the infected leaf extracts in Western blots and double-antibody sandwich ELISA. The viral RNA was used as a template for a cDNA library, and nucleotide sequence analysis identified cloned cDNAs representing sequences corresponding to the three Tospovirus genome segments. Sequence comparisons showed that the new virus had the highest similarity to Chrysanthemum stem necrosis virus (CSNV). Phylogenetic analysis of two genome regions confirmed that this virus, provisionally named Melon severe mosaic virus (MeSMV), is a previously undescribed Tospovirus sp. belonging to the "new world" clade of Tospovirus spp. An initial survey of various cucurbit crops in various states of Mexico confirmed the widespread occurrence of this virus.

Molecular characterization and detection of plum bark necrosis stem pitting-associated virus.

The complete RNA genome of plum bark necrosis stem pitting-associated virus (PBNSPaV) was cloned and sequenced and was determined to be 14, 214 nts long. The genome structure revealed seven major open reading frames (ORFs), and nontranslated regions at the 5' and 3' ends. PBNSPaV represents the simplest genome organization in the genus Ampelovirus, family Closteroviridae. The ORFs 1a and 1b encode, respectively, a large polyprotein with a molecular mass (Mr) of 259.6 kDa containing conserved domains characteristic of a papain-like protease, methyltransferase and helicase (ORF1a) and a 64.1-kDa protein of eight conserved motifs characteristic of viral RNA-dependent RNA polymerase (RdRp) (ORF1b). ORF1b is presumably expressed via a +1 ribosomal frameshift mechanism. ORF2 encodes a small 6.3-kDa hydrophobic protein of unknown function. ORF3 encodes a 57.4-kDa protein, a homologue of the HSP70 family of heat shock proteins. ORF4 encodes a 61.6-kDa protein with unknown function. ORF5 encodes a 35.9-kDa capsid protein (CP). Lastly, ORF6 encodes a 25.2-kDa minor capsid protein (CPm). Phylogenetic analyses performed on sequences of the HSP70h RdRp and CP support classification of the virus in the genus Ampelovirus. A real-time TaqMan RT-PCR assay and a one-step RT-PCR were developed for PBNSPaV detection and compared using three different sample preparation methods.

Measurement of talent in volleyball: 15-month follow-up of elite adolescent players.

AIM: The purpose of this study was two-fold: first, to examine the contribution of a battery of physical and motor tests to early phases of talent detection and early development in volleyball, and second, to differentiate between and compare the motor ability of 16-year-old starter (S) and non-starter (NS) volleyball players. METHODS: Fifteen male adolescent volleyball players underwent assessment of physical and motor ability 6 times during a 15-month training program; however, not all of them took part in each testing phase. The battery was composed of 8 physical and motor tests and 2 skill tests. The physical and motor tests included 2 speed tests, an agility run, 4 explosive power tests, and an endurance test. The skill tests evaluated service accuracy at rest and following effort. RESULTS: All participants improved their results in all but 2 tests (endurance and skill tests) across testing phases. Comparisons between the S (n=8) and NS (n=7) revealed that only one physical explosive power test (vertical jump with approach), was found to be a good indicator for distinguishing between the 2 groups of players. CONCLUSION: It was concluded that the volleyball battery of tests was not sensitive enough to distinguish between the ''good'' and ''very good'' players suggesting that physical and motor tests do not reflect open skill ability in volleyball.

A cumulative effect of physical training on bone strength in males.

Weight-bearing, high-impact exercise, as opposed to nonimpact exercise, has been demonstrated to increase bone mineral density. This was traditionally demonstrated with dual energy X-ray absorptiometry. Our objective was to assess the differences in bone properties, using quantitative ultrasound (QUS, Sunlight Omnisense, Sunlight Medical, Ltd., Tel Aviv, Israel), in male athletes involved in a weight-bearing, impact sport (soccer, SC) or a nonimpact sport (swimming and water polo, AQ), compared with nonathletic control (C) males. A total of 266 boys and men, aged 8 - 23 years, were divided into children (11.1 +/- 1.0 years; 34 SC, 34 AQ, 25 C), adolescents (14.7 +/- 1.2 years; 32 SC, 31 AQ, 31 C), and young adults (19.8 +/- 1.1 years; 31 SC, 24 AQ, 24 C) . Training experience varied between 1.5 years in the children to 15 years in the adults. Bone speed of sound (SOS) was measured bilaterally at the distal radius and the mid-tibia. Body fat was significantly lower in athletes compared with C. AQ were generally heavier and had a higher fat-free mass compared with SC and C, with no significant differences in height between groups. Radial SOS increased with age, but no differences were observed between activity groups or between the dominant (D) and nondominant (ND) arm. Tibial SOS also increased with age. In the children and adolescents, no differences were observed between activity groups. However, among adults, both SC and AQ had higher tibial SOS compared with C. These differences were mainly explained by differences in fat-free mass. Among young adults but not among children and adolescent males, both soccer and aquatic sports appear to be associated with higher bone SOS in the lower, but not the upper, extremities. Further studies are needed to assess possible sport-specific mechanisms which affect bone properties and to determine the minimal cumulative effect which is needed to influence bone properties.