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Pacific bluefin tuna (PBT), Thunnus orientalis, is one of the most important species for aquaculture in Japan. Recently, the reduction in muscle fat content associated with sexual maturation in farmed PBT has become a serious problem. To develop technologies for inducing sterility, detailed and reliable data on gonadal development in PBT are needed. Here, we demonstrated the process of gonadal sex differentiation, and of early ovarian and testicular development during the immature stages in PBT. Gonadal sex differentiation was first characterized by the formation of the ovarian cavity in female and of the efferent ducts in male 57 days post hatching (dph). The gonads then differentiated into ovaries or testes according to the genotypic sex until 83 dph. During this period, primordial germ cells, oogonia, and type-A spermatogonia were solitarily distributed in the gonads, and the number of germ cells did not differ between sexes. After gonadal sex differentiation, gonads of PBTs developed in a sexually dimorphic manner: proliferation and differentiation of germ cells occurred earlier in the ovaries than in the testes. The oogonia in ovaries formed cysts at 185 dph, but the type-A spermatogonia were solitarily distributed in testes at this stage, and cysts of type-A spermatogonia were first observed at 247 dph. Moreover, the oogonia entered meiosis and differentiated into chromatin-nucleolus stage oocytes until 247 dph, and subsequently into peri-nucleolus stage oocytes until 285 dph, whereas the type-A spermatogonia differentiated into type-B spermatogonia, spermatocytes, spermatids, and spermatozoa from 446 dph onwards. We believe the results of this study provide the necessary basis for future studies on sterile PBT production.

BackgroundEvidence-based international expert consensus regarding the impact of peripheral nerve block (PNB) use in total hip/knee arthroplasty surgery.MethodsA systematic review and meta-analysis: randomized controlled and observational studies investigating the impact of PNB utilization on major complications, including mortality, cardiac, pulmonary, gastrointestinal, renal, thromboembolic, neurologic, infectious, and bleeding complications.Medline, PubMed, Embase, and Cochrane Library including Cochrane Database of Systematic Reviews, Cochrane Central Register of Controlled Trials, NHS Economic Evaluation Database, were queried from 1946 to August 4, 2020.The Grading of Recommendations Assessment, Development, and Evaluation approach was used to assess evidence quality and for the development of recommendations.ResultsAnalysis of 122 studies revealed that PNB use (compared with no use) was associated with lower ORs for (OR with 95% CIs) for numerous complications (total hip and knee arthroplasties (THA/TKA), respectively): cognitive dysfunction (OR 0.30, 95% CI 0.17 to 0.53/OR 0.52, 95% CI 0.34 to 0.80), respiratory failure (OR 0.36, 95% CI 0.17 to 0.74/OR 0.37, 95% CI 0.18 to 0.75), cardiac complications (OR 0.84, 95% CI 0.76 to 0.93/OR 0.83, 95% CI 0.79 to 0.86), surgical site infections (OR 0.55 95% CI 0.47 to 0.64/OR 0.86 95% CI 0.80 to 0.91), thromboembolism (OR 0.74, 95% CI 0.58 to 0.96/OR 0.90, 95% CI 0.84 to 0.96) and blood transfusion (OR 0.84, 95% CI 0.83 to 0.86/OR 0.91, 95% CI 0.90 to 0.92).ConclusionsBased on the current body of evidence, the consensus group recommends PNB use in THA/TKA for improved outcomes.Recommendation: PNB use is recommended for patients undergoing THA and TKA except when contraindications preclude their use. Furthermore, the alignment of provider skills and practice location resources needs to be ensured. Evidence level: moderate; recommendation: strong.

Kawakawa Euthynnus affinis, also known as eastern little tuna or mackerel tuna, is a species of tuna (Thunnini tribe, subgroup Pelagiaria clade) (Sanciangco et al., 2016). Tuna includes 15 species: eight of genus Thunnus (true tuna) and seven “tuna-like” of four genera: Allothunnus, Auxis, Euthynnus, and Katsuwonus. E. affinis is widely distributed throughout the tropical and subtropical waters of the continental shelf areas of the Indo-Pacific region (Collette, 2001). The fish reaches a length of 45–60 cm and matures at approximately 3 years of age. It inhabits almost exclusively the upper layers of the ocean (Bernal et al., 2017) and feeds mainly on small pelagic fish (Griffiths et al., 2009). E. affinis makes up a substantial proportion of the commercial and artisanal fishery in many countries of the Indo-Pacific region. The meat of E. affinis is of high quality (Mukundan et al., 1979) with a comparatively high level of docosahexaenoic acid (Saito et al., 1999), but deteriorates rapidly if not properly handled (Mukundan et al., 1979). E. affinis exhibits the swimming mechanics of true tunas (Donley and Dickson, 2000) but has no swim bladder and differs from true tunas in red muscle distribution, allometry, and vascular anatomy (Bernal et al., 2017). The ability to maintain an elevated temperature in eye, brain, and red muscle has been suggested for the genus Euthynnus (Dickson et al., 2000), but reports specific to E. affinis are lacking. Compared to true tunas, E. affinis has received scant attention from researchers, and little is known about its biology and physiology. This is likely to change, as E. affinis has recently become of interest in marine aquaculture. E. affinis is the second tuna species whose full-life cycle culture in captivity has been developed so far, including spawning, egg collection, incubation, larval rearing, and grow-out to marketable size (Yazawa et al., 2015, 2016). Aquaculture in general is currently facing significant challenges to increasing production while maintaining sustainability (Bridson et al., 2020). Genetic improvement, via selective breeding and genetic engineering, is a major focus of research and can yield rapid benefits to efficient production in fish farming (Lu and Luo, 2020). To these ends, a high-quality species genome assembly is critical. Despite recent advances in sequencing technologies and genomics that, in addition to basic fish science (Lien et al., 2016; Hughes et al., 2018; Yuan et al., 2018; Du et al., 2020), have applications to aquaculture practices (Lu and Luo, 2020) and fisheries (Benestan, 2020), genomic information of tuna species is limited. To date, the genomes of only three tuna species are available in the public repositories, none of which are assembled to chromosome level. This situation exists within the entire Pelagiaria clade that, along with tuna, includes the economically important mackerel (Scombrini tribe) and bonito (Sardini tribe). Here, we report the chromosome-level genome assembly of kawakawa E. affinis (NCBI:txid8227). To our knowledge, this is the first available chromosome-level assembly within the Pelagiaria clade. The reported genome assembly is accompanied by transcriptome assembly, genetic linkage map, annotation of transposons, repetitive elements, and 23,059 genes. The dataset provides a solid genome resource not only for further study of E. affinis basic biology and genome-scale selective breeding but also for enhancing both basic and applied research within the Pelagiaria clade.

Study design Systematic review. Objective To determine the difference in postoperative sagittal alignment when single level cervical radiculopathy or myelopathy is treated with an anterior cervical discectomy and fusion (ACDF) using a structural graft and plate, a stand-alone ACDF, or a total disc arthroplasty (TDR). We also wanted to determine if postoperative sagittal alignment impacted clinical outcomes in this patient population. Summary of background data Although there are several accepted techniques for interbody reconstruction during single level anterior cervical surgery, little is known on the impact of any of them on segmental and regional sagittal alignment. Methods A systematic review of the literature was performed according to the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines of the PubMed, Embase, and Cochrane databases. Only studies which contained pre and postoperative C2-7 Cobb angles as well as Neck Disability Index (NDI) scores following single level traditional ACDF with plate and cage, stand-alone ACDF, or TDR with at least 1 year follow up were included. Results There were 12 publications that fulfilled the inclusion criteria with a total of 658 patients. Cluster regression analysis showed no difference between treatment arms at each respective time-point or in the overall change in NDI from preoperative to postoperative (P = 0.88). Cluster regression analysis showed no difference between treatment arms at each respective time point or in the overall change in lordosis from pre-op to post-op (P = 0.93). Conclusion This review provides evidence that while NDI scores improved across all devices, there was no significant difference in NDI improvement or C2-7 Cobb angle change between single level traditional ACDF, stand-alone ACDF, or TDR. Although TDR has not been considered a lordosis producing operation, our investigation shows it does not differ significantly in sagittal profile from other cervical fusion techniques. Moreover, we show that the NDI score improved, regardless of device implanted. Level of evidence 1.

Examination of gonads, fresh coloration and external morphology of Parupeneus spilurus (Perciformes: Mullidae), based on 55 males [115.4–315.9 mm SL (mean 254.1 mm)] and 22 females [116.2–299.5 mm SL (225.3 mm)], provided evidence of both sexual dichromatism and dimorphism. Males consistently had six to nine oblique yellow stripes and two yellow stripes on the second dorsal fin and anal fin, respectively. Females less than 212.3 mm SL had a reddish second dorsal fin, becoming yellowish (but no stripes or blotches) with maturity (> 215.4 mm SL). The anal fin of females was usually reddish without yellow stripes, even at full maturity, although a single yellow stripe on the anal fin is likely during breeding periods (at least before ovulation). Whereas the caudal fin of females always lacked yellow markings, these occurred in mature males (> 193.0 mm SL), tending to change from stripes to blotches via broken stripes with subsequent growth. Males differed slightly from females in some relative proportions (orbit diameter, and upper-jaw and anal-fin base lengths). Parupeneus spilurus was found to be a gonochoristic, multiple-spawning species (more than one spawning per year) with asynchronous ovarian development.

In bluefin tuna aquaculture, high mortalities of hatchery-reared juveniles occur in sea cages owing to wall collisions that are caused by high-speed swimming in panic due to changes in illuminance. Here, we report that targeted gene mutagenesis of the ryanodine receptor (RyR1b), which allows the sarcoplasmic reticulum to release Ca2+ in fast skeletal muscle, using highly active Platinum TALENs caused slow swimming behaviour in response to external stimuli in Pacific bluefin tuna (PBT) larvae. This characteristic would be a useful trait to prevent wall collisions in aquaculture production. A pair of Platinum TALENs targeting exons 2 and 43 of the PBT ryr1b gene induced deletions in each TALEN target site of the injected embryos with extremely high efficiency. In addition, ryr1b expression was significantly decreased in the mutated G0 larvae at 7 days after hatching (DAH). A touch-evoked escape behaviour assay revealed that the ryr1b-mutated PBT larvae swam away much less efficiently in response to mechanosensory stimulation at 7 DAH than did the wild-type larvae. Our results demonstrate that genome editing technologies are effective tools for determining the functional characterization of genes in a comparatively short period, and create avenues for facilitating genetic studies and breeding of bluefin tuna species.

Background: Phenomena including early sport specialization and year-round training and competition have contributed to an increase in pediatric sports injuries. There has been a concomitant increase in clinical studies focusing on physically active children and adolescents. These studies include investigations of patient-reported outcome measures (PROMs). While the use of PROMs in pediatric orthopaedics has been increasing, PROMs are often inappropriately applied to study populations for whom they are not specifically validated. Purpose: The purpose of this study was to establish a comprehensive list of pediatric- and adolescent-validated PROMs and catalog their psychometric properties as a resource for clinicians and researchers. Study Design: Systematic review; Level of evidence, 4. Methods: A systematic review of articles in PubMed, Embase, and the Cochrane library was performed to identify articles developing and validating PROMs appropriate for use in pediatric sports medicine research. The inclusion criteria were as follows: age Results: Our search strategy identified 14,708 unique articles, among which 139 studies (0.9%) were included in the final data analysis. Forty-seven distinct PROMs were identified, as well as 160 cross-cultural adaptations. While all identified PROMs were validated in physically active young participants, only 12 (26%) were specifically created initially for active children. Thirty (64%) PROMs were health-related quality-of-life measures; 13 (28%) were psychosocial measures; and 4 (9%) were activity scales. No studies validated PROMs for use with wrist/hand injuries, and only 1 PROM each was valid for hip, back/spinal, and foot/ankle injuries in pediatric sports. Conclusion: This systematic review yielded 47 unique PROMs reliable and valid for use in pediatric and adolescent sports medicine. This list will unify clinicians and researchers in using these age-appropriate measures while identifying areas that are still in need of appropriate PROMs for young athletes.

Patellofemoral arthroplasty (PFA) and total knee arthroplasty (TKA) are accepted treatments for end-stage isolated patellofemoral osteoarthritis (PFOA). However, complications and re-operations have historically differed between the two procedures. We performed a systematic review to report on the re-operation rates between TKA and modern PFA for isolated PFOA. Systematically identified publications reporting on patients that underwent either TKA or modern PFA for isolated PFOA were reviewed. Meta-analysis software was used to screen potential articles with at least one year follow-up that detailed reasons for re-operation. Data was extracted and analyzed for all re-operations. Survival of the implant was used as the primary outcome; return to the operating room (OR) for any reason was used as a secondary outcome. The weighted rate of either conversion or revision arthroplasty in the PFA group and the TKA group was 6.34 and 0.11, respectively. The weighted rate of return to the OR for bony and soft tissue procedures was 1.06 and 0.79, respectively. The weighted rate of manipulation under anaesthesia (MUA) was 0.32 and 1.23, respectively. Patients who undergo PFA may be more likely to return to the operating room for conversion to TKA and/or revision surgery than those who undergo TKA.

Background Slipped capital femoral epiphysis (SCFE) is an important cause of hip pain and disability in pediatric patients. SCFE occurs bilaterally in 12% to 80% of cases, and the risk of contralateral SCFE is noted to be 2335 times higher than the index SCFE. Several studies have reported risk factors for contralateral SCFE; however, these studies have not been systematically analyzed. The purpose of this systematic review and meta-analysis was to review and analyze risk factors for subsequent contralateral SCFE and identify the strongest risk factors for a subsequent slip. Methods A systematic review was performed of all observational studies focusing on risk factors for subsequent contralateral SCFE indexed in Medline, Embase, and Cochrane databases. Data extraction was performed and summarized using descriptive statistics. Meta-analysis was performed for risk factors with sufficient constituent study data. Quality assessment was performed using the Newcastle-Ottawa Scale, and funnel plots were generated to assess publication bias. Results The initial search strategy identified 226 references, and after exclusions, 20 studies were included in this analysis. Demographic risk factors included age, sex, weight, body mass index, ethnicity, and urban/rural residence; clinical risk factors included endocrine abnormality, duration of symptoms, slip stability, and slip chronicity; and radiographic risk factors included slip angle, triradiate cartilage, alpha angle, posterior sloping angle (PSA), physeal sloping angle, modified Oxford score, and bone age. Younger patient age, body mass index≥95th percentile, presence of an endocrine abnormality, higher PSA of the unaffected hip, and lower modified Oxford score have been noted to be significant risk factors for contralateral SCFE. Meta-analysis showed that younger age (-0.9; confidence interval, -1.1, -0.6), and higher PSA (4.7 degrees; 95% confidence interval, 3.3-6.2 degrees) of the unaffected hip were predictive of subsequent contralateral SCFE. The majority of studies were of good quality. Conclusion There are several risk factors for subsequent contralateral SCFE. On the basis of the available data, younger patients with a high PSA of the unaffected hip would most likely benefit from prophylactic fixation of the unaffected hip. Level of evidence Level II.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

The technologies of developmental biology have been improving over the past 30 years. Full genome sequencing enables between species comparisons, and genomics have revealed continuous genetic relationships between species. Genetic manipulation, such as modification of specific genes and genetic improvements, has been getting easier by gene editing technology, if fresh fertilized fish eggs are prepared and cultured. On the other hand, micro-manipulation, such as micro-operation of embryos, also opens a new possibility of surrogate production. Gametes have been induced through germ-line chimera induced artificially by germ-cell transplantation between different species. In this context, the intercellular interaction between species with different genomes is an important subject to be analyzed. In this section, general methods are described for artificial fertilization and cultivation in teleosts, and a few methods useful for experimental embryology are introduced. Then, observation procedures and development are described in several species. Finally, some experiments are introduced to study embryonic mechanisms in freshwater species. These experiments may be possible in other species if chorion can be removed at the early embryonic stage.

Objective.Total hip replacement (THA) surgery is a successful procedure, yet blacks in the United States undergo THA less often and reflect poorer outcomes than whites. The purpose of this study is to systematically review the literature on health-related quality of life after THA, comparing blacks and whites.Methods.A librarian-assisted search was performed in Medline through PubMed, Embase, and Cochrane Library on February 27, 2017. Original cohort studies examining pain, function, and satisfaction in blacks and whites 1 year after elective THA were included. Using the Patient/Population–Intervention–Comparison/Comparator–Outcome (PICO) process format, our population of interest was US black adults, our intervention was elective THA, our comparator was white adults, and our outcomes of interest were pain, function, and satisfaction after elective THA. The protocol was registered under the PROSPERO international register, and the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) guidelines were followed.Results.Of the articles, 4739 were screened by title, 180 by abstract, 25 by full text, and 4 remained for analysis. The studies represented 1588 THA patients, of whom 240 (15%) were black. All studies noted more pain and worse function for blacks; although differences were statistically significant, they were not clinically significant. One study sought and identified less satisfaction for blacks after THA, and 1 study showed worse fear and anxiety scores in blacks.Conclusion.When measured, there are small differences in THA outcomes between blacks and whites, but most studies do not analyze/collect race. Future studies should address the effect of race and socioeconomic factors on healthcare disparities.

This book introduces the reader to the very sensitive, emotional, and sometimes intimidating topic of death and dying. It shows the reader how to communicate their decisions about their end-of-life...

Background: Reportedly 2% to 5.7% of total knee arthroplasties (TKAs) require revision within 5 years. The purpose of this study was to determine whether blacks are at higher risk of TKA revision than whites in the United States. Methods: We performed a systematic review of English-language articles published from 2000 to 2015. Study inclusion criteria were (1) performance of the study in the United States, (2) TKA as the primary procedure studied, (3) a follow-up period at least 2 years, (4) reporting of revision rates, and (5) analysis of patient race as an independent predictor of revision. We then performed a random-effects meta-analysis to calculate a pooled hazard ratio for TKA revision in blacks compared with whites. Results: A total of 4,286 studies were identified and screened by title; 106, by abstract; and 24, by full text. Six studies met the inclusion criteria. Only 4 of the 6 studies could undergo meta-analysis because of overlapping study populations in 3 of them. The meta-analysis represented 451,960 patients who underwent TKA, of whom 28,772 (6.4%) were black. Of the total, 31,568 patients (7.0%) underwent revision surgery. The risk of revision TKA was significantly higher among blacks than whites (pooled hazard ratio, 1.38; 95% confidence interval, 1.20 to 1.58; p < 0.001). Analysis of the 3 studies with overlapping study populations demonstrated discordant results as a result of adjustment compared with non-adjustment for insurance eligibility, a surrogate for socioeconomic status. Conclusions: Blacks in the United States are at higher risk of revision TKA than whites. Socioeconomic status contributed to revision risk and is an important confounder in analyses of race. Level of Evidence: Prognostic Level III. See Instructions for Authors for a complete description of levels of evidence.

Objective:The current study assessed changes in children and mothers’ nutritional status before and after raising Bangladeshi households out of extreme poverty through an income-generating activities (IGA) programme.Design:Extreme poor households took part in the IGA programme for 2 years and recruitment took place over four waves in annual cycles. Children and mothers were measured with regarding their nutritional status before and after the IGA programme commenced.Settings:Rural Bangladesh.Subjects:Three-hundred and eighty-two children under 5 years of age at recruitment, and their mothers.Results:After 2 years of the IGA programme, the prevalence of stunting significantly declined from 40·3 % to 33·0 % (P = 0·003), anaemia declined from 51·6 % to 44·0 % (P = 0·020) while mothers’ CED (Chronic Energy Deficiency) declined from 52·0 % to 42·7 % (P < 0·001), but no significant changes were found in children’s wasting, declining from 25·4 % to 21·5 %, underweight which remained the same at 43·2 %, while mothers’ anaemia rose from 39·3 % to 42·7 %. There were also highly significant improvements in household socio-economic status. Increases in socio-economic security (especially in relation to cash savings and net income) and improvements in food quantity and quality (indicated by greater food diversity and animal food intake) were associated with normal nutritional status, and cessation of open defecation was associated with reduction in mothers’ and child anaemia.Conclusion:The IGA programme was associated with increased household socio-economic security, such as asset accumulation, food security and sanitation, and with improvements in the nutritional status of children and their mothers in extreme poor households.

Systematic review.To determine the difference in postoperative sagittal alignment when single level cervical radiculopathy or myelopathy is treated with an anterior cervical discectomy and fusion (ACDF) using a structural graft and plate, a stand-alone ACDF, or a total disc arthroplasty (TDR). We also wanted to determine if postoperative sagittal alignment impacted clinical outcomes in this patient population.Although there are several accepted techniques for interbody reconstruction during single level anterior cervical surgery, little is known on the impact of any of them on segmental and regional sagittal alignment.A systematic review of the literature was performed according to the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines of the PubMed, Embase, and Cochrane databases. Only studies which contained pre and postoperative C2-7 Cobb angles as well as Neck Disability Index (NDI) scores following single level traditional ACDF with plate and cage, stand-alone ACDF, or TDR with at least 1 year follow up were included.There were 12 publications that fulfilled the inclusion criteria with a total of 658 patients. Cluster regression analysis showed no difference between treatment arms at each respective time-point or in the overall change in NDI from preoperative to postoperative (P = 0.88). Cluster regression analysis showed no difference between treatment arms at each respective time point or in the overall change in lordosis from pre-op to post-op (P = 0.93).This review provides evidence that while NDI scores improved across all devices, there was no significant difference in NDI improvement or C2-7 Cobb angle change between single level traditional ACDF, stand-alone ACDF, or TDR. Although TDR has not been considered a lordosis producing operation, our investigation shows it does not differ significantly in sagittal profile from other cervical fusion techniques. Moreover, we show that the NDI score improved, regardless of device implanted.1.

This paper contributes to two strands of literature: disaster literature on post-disaster mental health and economics literature on migration. Remittances are a sizable source of income in many developing countries. Evidence suggests, however, that when adults migrate in order to support their families via remittances, family members left behind often experience poorer physical and mental health. We study the effects of remittances on the mental health of victims of a disaster, the earthquake (EQ) that hit Nepal in 2015. We used three waves of data from 335 individuals in 6 villages in Western Nepal in which emigration is prevalent. The first wave of the survey was conducted one year before the 2015 EQ. In the third wave, which was conducted one year after the EQ, the respondents aged 15 and older were assessed for post-traumatic stress disorder (PTSD) and depressive symptoms. PTSD symptoms were evaluated by the 17-item Checklist Civilian (PCL-C), and depressive symptoms were measured by the 20-question Epidemiologic Studies Depression Scales (CESD) questionnaire. In order to avoid capturing the impacts of remittances for daily expenses and reduce possible endogeneity in the remittance variables, we measured the change in remittances following the EQ. After controlling for pre-disaster body-mass index and asset holdings, we found that the increase in remittances sent to HHs reduced psychological distress measured by the PTSD and depression severity scores. The remittance variables, however, did not alleviate mental disorder as defined by the international-standard cutoff points of PCL-C and CES-D.

Surrogate propagation is a systematic approach to producing donor-derived gametes using germline chimeras. In fish, the use of germline chimeras to study the development of germ cells was first conducted in the 1990s in the model fish species medaka (Oryzias latipes) and zebrafish (Danio rerio). More recently, surrogate propagation has been actively investigated as a means of efficient gamete production not only in model fish species but also in aquaculture species and endangered species. Surrogate propagation has the following components: combination of the donor and host species, host sterilization, donor cell preparation, transplantation of germ cells, and gametogenesis and gamete production in surrogate fish. In this review, we first provide a general overview of previous studies related to germ cell transplantation and the methodologies developed for different species, and consider how these have been applied in practice. Second, we consider the development of primordial germ cells in fish embryos, particularly the molecular biological approaches used for the visualization of germ cells and sterilization of host embryos. Finally, we discuss sex control in germline chimeras, which may be a key component of the use of surrogate production in aquaculture. We focus on techniques to produce sterile fish, as these are crucial to the exclusive production of donor gametes in a surrogate host. The advantages and disadvantages of various aspects of surrogate propagation are discussed, the potential use of surrogate propagation as a seedling production system is considered, and future perspectives for aquaculture are suggested.

Osteoporosis in axial spondyloarthritis may be modified by therapy. The purpose of this systematic review is to describe (i) the effect of TNFi on BMD, (ii) the effect of secukinumab on BMD, and (iii) the effect of secukinumab on radiographic disease progression in axSpA. We searched PubMed, Embase, and Cochrane using the following retrieval languages: spondyloarthritis, ankylosing spondylitis, TNF, IL-17, x-rays, and osteoporosis. Twenty-nine studies were included; 27 re: TNFi and BMD, and 2 re: IL-17 blockers and x-ray progression. TNFi over 2–4 years increased BMD of the lumbar spine (3.2–14.9%) and hip (2.26–4.7%) without reducing vertebral fractures. Secukinumab reduced radiographic progression; none (73%) and minimal (79%) at 4 years. No data on IL-17 blockade and bone were found. TNFi therapy improves bone density but not vertebral fracture rates. Secukinumab improves symptoms and may slow radiographic progression. Data is lacking regarding the effects of secukinumab on BMD and fractures. These are important questions which may impact the choice of therapy.

Background Evidence-based international expert consensus regarding anaesthetic practice in hip/knee arthroplasty surgery is needed for improved healthcare outcomes. Methods The International Consensus on Anaesthesia-Related Outcomes after Surgery group (ICAROS) systematic review, including randomised controlled and observational studies comparing neuraxial to general anaesthesia regarding major complications, including mortality, cardiac, pulmonary, gastrointestinal, renal, genitourinary, thromboembolic, neurological, infectious, and bleeding complications. Medline, PubMed, Embase, and Cochrane Library including Cochrane Database of Systematic Reviews, Cochrane Central Register of Controlled Trials, NHS Economic Evaluation Database, from 1946 to May 17, 2018 were queried. Meta-analysis and Grading of Recommendations Assessment, Development and Evaluation approach was utilised to assess evidence quality and to develop recommendations. Results The analysis of 94 studies revealed that neuraxial anaesthesia was associated with lower odds or no difference in virtually all reported complications, except for urinary retention. Excerpt of complications for neuraxial vs general anaesthesia in hip/knee arthroplasty, respectively: mortality odds ratio (OR): 0.67, 95% confidence interval (CI): 0.57–0.80/OR: 0.83, 95% CI: 0.60–1.15; pulmonary OR: 0.65, 95% CI: 0.52–0.80/OR: 0.69, 95% CI: 0.58–0.81; acute renal failure OR: 0.69, 95% CI: 0.59–0.81/OR: 0.73, 95% CI: 0.65–0.82; deep venous thrombosis OR: 0.52, 95% CI: 0.42–0.65/OR: 0.77, 95% CI: 0.64–0.93; infections OR: 0.73, 95% CI: 0.67–0.79/OR: 0.80, 95% CI: 0.76–0.85; and blood transfusion OR: 0.85, 95% CI: 0.82–0.89/OR: 0.84, 95% CI: 0.82–0.87. Conclusions Recommendation: primary neuraxial anaesthesia is preferred for knee arthroplasty, given several positive postoperative outcome benefits; evidence level: low, weak recommendation. Recommendation: neuraxial anaesthesia is recommended for hip arthroplasty given associated outcome benefits; evidence level: moderate-low, strong recommendation. Based on current evidence, the consensus group recommends neuraxial over general anaesthesia for hip/knee arthroplasty. Trial registry number PROSPERO CRD42018099935.

The induction of germ-line chimerism in fish is a strategy for the reproduction of endangered or genetically valuable fish species. Chimeras can be created by transplanting a single primordial germ cell or multiple blastomeres from a donor into a sterile host embryo. When the host reaches sexual maturity, it will produce donor-originating gametes throughout its reproductive life span. This technique provides unique experimental conditions for basic biology research in model fish species like zebrafish. The success of cell transplantation relies on the effective sterilization of host embryos, the correct identification of developing germ cells, and the synchronization of migratory cues between the host and the transplanted cells. Developments in non-transgenic methods of germ cell ablation and identification have made germ cell transplantation more applicable to use in conservation and aquaculture. In this chapter, we provide a protocol for germ cell labeling by injection of chimeric RNA or FITC-dextran, the sterilization of host embryos using an antisense morpholino oligonucleotide, and two methods for producing germ-line chimeras in zebrafish: single primordial germ cell transplant and blastomere transplant.

Bluefin tuna is one of the most important aquaculture species in several countries; however, information regarding the primordial germ cell (PGC) development and migration in this species is scarce. This information is vital for application in reproductive biotechnology, for example, induced sterility through targeted cell ablation or PGC manipulation. Teleost PGC can be visualized by injecting an RNA transcribed from the fusion product of a fluorescent protein gene attached to the 3' untranslated region (3'UTR) of zebrafish nanos3 (zf-nos3) into eggs. In this study, we identified the PGC and its migratory pathway during early embryogenesis and larvae development by injecting the GFP-zf-nos3 3'UTR mRNA into the Pacific bluefin tuna (PBT, Thunnus orientalis). PBT PGCs were initially found around the marginal and dorsal regions of the blastodisc at 50%-epiboly stage. The PGCs were aligned as two elongated lines at the posterior part of the embryonic body during the early segmentation period, and eventually formed a single tight cluster underneath somites 10 to 15 of the embryonic body until the late segmentation period. Although the aggregated PGCs stayed at the same position during hatching, they started migrating anteriorly and were split into two populations at 3 days after hatching (DAH). Until 15 DAH, these PGCs settled in two bilateral lines at the apex of the peritoneal cavity. Histological analysis of PBT larvae revealed that at 3 and 5 DAH, the PGCs were not enclosed by the somatic cells, whereas at 15 DAH, they were entirely covered by the somatic cells, indicating the development of the primordial gonads. These results are essential for future experiments in germ line control technologies for bluefin tuna.

The induction of germ-line chimerism in fish is a strategy for the reproduction of endangered or genetically valuable fish species. Chimeras can be created by transplanting a single primordial germ cell or multiple blastomeres from a donor into a sterile host embryo. When the host reaches sexual maturity, it will produce donor-originating gametes throughout its reproductive life span. This technique provides unique experimental conditions for basic biology research in model fish species like zebrafish. The success of cell transplantation relies on the effective sterilization of host embryos, the correct identification of developing germ cells, and the synchronization of migratory cues between the host and the transplanted cells. Developments in non-transgenic methods of germ cell ablation and identification have made germ cell transplantation more applicable to use in conservation and aquaculture. In this chapter, we provide a protocol for germ cell labeling by injection of chimeric RNA or FITC-dextran, the sterilization of host embryos using an antisense morpholino oligonucleotide, and two methods for producing germ-line chimeras in zebrafish: single primordial germ cell transplant and blastomere transplant.

SummaryThe nutritional status of under-five-year-old children is a sensitive indicator of a country’s health status as well as economic condition. The objectives of this study were to analyse trends in the nutritional status in Bangladeshi children over the period 1996–2007 and to examine the associations between nutritional and socioeconomic status variables. Bangladesh Demographic Health Surveys (BDHS) were the source of data, and a total of 16,278 children were examined. The Z-scores of the children were analysed as continuous as well as categorical variables (stunted, underweight and wasted). The socioeconomic status variables used were region, urban–rural residence, education and occupation of the parents, house type and household possession score. A series of General Linear Model and Sequential Linear and Binary Logistic Regression analyses were done to assess the relationship between demographic and socioeconomic variables and nutritional status. The trends of Z-scores were analysed by survey, as well as by child birth cohort. Region, house type, educational level of parents and household possession score showed significant associations with all three Z-scores of children after removing the effects of age, period of DHS and other explanatory variables in the model. No significant sex difference was observed between any of the Z-scores. There were improvements in mean WAZ and HAZ between 1996 and 2007 but deterioration in mean WHZ over this period. The obesity rate was below 2% in 2007, although the absolute numbers of obese children had nearly doubled in this 12-year period. Children from poorer households showed greater improvement than their better-off counterparts. The study reveals that over the years there has been substantial improvement in nutritional status of under-five children in Bangladesh and the main gains have been amongst the lower socioeconomic groups; it is also evident that malnutrition in Bangladesh is a multidimensional problem, like poverty itself, and warrants a proper policy mix and programme intervention.

Microinjection is a powerful tool for studying embryonic development and analyzing gene functions in fish. This technique was first applied to model species of fish such as zebrafish and medaka whose egg chorions could be removed or softened before microinjection. Recent progress in genome editing using TALEN and CRISPR has opened the opportunity to analyze gene functions in a much wider range of fish including those important to marine aquaculture. Therefore, application of the microinjection technique is also required in these species. However, the characteristics of fish eggs vary widely between species and several technical difficulties need to be overcome in order to use microinjection in a wider range of species. To obtain consistent results with microinjection, an optimal method has to be developed for each target species. In this chapter, we describe the physical characteristics of the eggs of fish species that have been used in microinjection experiments in our laboratory and detail the microinjection system we developed for fish eggs with a hard chorion, such as those of marine species.

Microinjection is a powerful tool for studying embryonic development and analyzing gene functions in fish. This technique was first applied to model species of fish such as zebrafish and medaka whose egg chorions could be removed or softened before microinjection. Recent progress in genome editing using TALEN and CRISPR has opened the opportunity to analyze gene functions in a much wider range of fish including those important to marine aquaculture. Therefore, application of the microinjection technique is also required in these species. However, the characteristics of fish eggs vary widely between species and several technical difficulties need to be overcome in order to use microinjection in a wider range of species. To obtain consistent results with microinjection, an optimal method has to be developed for each target species. In this chapter, we describe the physical characteristics of the eggs of fish species that have been used in microinjection experiments in our laboratory and detail the microinjection system we developed for fish eggs with a hard chorion, such as those of marine species.

BACKGROUND Transforaminal lumbar interbody fusion (TLIF) is a well-accepted surgical technique for the treatment of degenerative spinal conditions and spinal deformity. The TLIF procedure can be performed open or using minimally invasive techniques. While several studies have found that minimally invasive TLIF (MI-TLIF) has advantages over open TLIF procedures with less blood loss, postoperative pain and hospital length of stay, opponents of the minimally invasive technique cite the lack of restoration of lumbar lordosis as a major drawback. With the increasing awareness of restoring sagittal alignment parameters in degenerative and deformity procedures, surgeons should understand the capabilities of different procedures to achieve surgical goals. To our knowledge, few studies have specifically studied the radiographic restoration of lumbar lordosis after MI-TLIF procedures. The purpose of this study was to perform a systematic review of the literature describing the sagittal lumbar radiographic parameter changes after MI-TLIF. METHODS Following PRISMA guidelines, systematic review was performed. With the assistance of a medical librarian, a highly-sensitive search strategy formulated on 1/19/2018 utilized the following search terms: “minimally invasive procedures,” “transforaminal lumbar interbody fusion,” “lumbar interbody fusion,” “diagnostic imaging,” “radiographs,” “radiography,” “x-rays,” “lordosis,” “lumbar vertebrae,” “treatment,” “outcome,” and “lumbosacral” using Boolean operators ‘AND’ and ‘OR’. Three databases were searched (PubMed/Medline, Embase, and Cochrane Library). An online system ( www.covidence.org ) was used to standardize article review. All studies were independently analyzed by two investigators and discrepancies mitigated by a third reviewer. Study selection for each cycle was Yes/No/Maybe. Cycles were: (1) (Title/Abstract); (2) (Full Text); (3) (Extraction). Inclusion criteria were: (1) All study designs, (2) MI-TLIF procedures, (3) Reporting total lumbar lordosis (LL) and/or segmental lordosis (SL) pre- and postoperatively. Exclusion criteria were: (1) non MI-TLIF procedures (ALIF, XLIF, LLIF, conventional TLIF, OLIF), (2) No reported LL or SL. RESULTS The search yielded 4,036 results with 836 duplicates leaving 3,200 studies for review. Cycle 1 eliminated 3,153 studies as irrelevant, thus, 47 were eligible for full-text review. Cycle 2 excluded 31 studies for No English full text (9), Oral/Poster (8), Wrong intervention/outcome (10), Duplicate listing (2), Full text not available (1), Literature review (1) resulting in 16 included studies. Study designs were: Randomized-controlled trial (3), Case series (6) and Retrospective (7). Mean # of subjects were 32.0 (range 8–95). Weighted-mean LL was 39.6°±9.2 (range 28–57) and postoperative LL was 45.0°±7.4 (range 36–67) with a LL post-pre difference of 5.2°±5.9 (range −7 to 15). Weighted-mean preoperative SL was 12.7°±4.3 (range 5–21) and postoperative SL was 15.0°±4.5 (range 5–22) with a SL post-pre difference of 2.1°±1.7 (range 0–8). CONCLUSIONS The current literature on MI-TLIF and restoration of LL/SL is limited to 16 published studies, 44% of which are retrospective. The published evidence supporting LL and SL restoration with MI-TLIF is sparse with variable results. This systematic review demonstrates the need for future high-level studies to fully elucidate the capabilities of MI-TLIF procedures for restoring lumbar and segmental lordosis.

ObjectiveTo analyse trends in maternal nutritional status in Bangladesh over a 12-year period and to examine the associations between nutritional status and socio-economic variables.DesignMaternal nutritional status indicators were height, weight and BMI. Socio-economic variables used were region, residency, education and occupation of the mothers and their husbands, house type, and possession score in the household.SettingBangladesh Demographic and Health Surveys (1996, 2000, 2004 and 2007) were the source of data.SubjectsA total of 16 278 mothers were included.ResultsAll of the socio-economic variables showed significant associations with maternal nutritional status indicators. Regional variation was found to be present; all three indicators were found to be lowest in the Sylhet division. Upward trends in maternal height, weight and BMI were evident from no possessions to four possessions in households, and for no education to higher education of women and their husbands. Bangladeshi mothers measured in 2007 were found to be on average 0·34 cm taller and 3·36 kg heavier than mothers measured in 1996. Between 1996 and 2007 maternal underweight fell from nearly 50 % to just over 30 % while overweight and obesity increased from about 3 % to over 9 % (WHO cut-offs) or from 7 % to nearly 18 % (Asian cut-offs).ConclusionsThe study reveals that over the 12-year period in Bangladesh there has been a substantial reduction in maternal underweight accompanied by a considerable increase in obesity. It is also evident that malnutrition in Bangladesh is a multidimensional problem that warrants a proper policy mix and programme intervention.

Background Clinical decision rules (CDRs) for pulmonary embolism (PE) have been validated in outpatients, but their performance in hospitalized patients is not well characterized. Objectives The goal of this systematic literature review was to assess the performance of CDRs for PE in hospitalized patients. Methods We performed a structured literature search using Medline, EMBASE and the Cochrane library for articles published on or before January 18, 2017. Two authors reviewed all titles, abstracts and full texts. We selected prospective studies of symptomatic hospitalized patients in which a CDR was used to estimate the likelihood of PE. The diagnosis of PE had to be confirmed using an accepted reference standard. Data on hospitalized patients were solicited from authors of studies in mixed populations of outpatients and hospitalized patients. Study characteristics, PE prevalence and CDR performance were extracted. The methodological quality of the studies was assessed using the QUADAS instrument. Results Twelve studies encompassing 3,942 hospitalized patients were included. Studies varied in methodology (randomized controlled trials and observational studies) and reference standards used. The pooled sensitivity of the modified Wells rule (cut-off ≤ 4) in hospitalized patients was 72.1% (95% confidence interval [CI], 63.7–79.2) and the pooled specificity was 62.2% (95% CI, 52.6–70.9). The modified Wells rule (cut-off ≤ 4) plus D-dimer testing had a pooled sensitivity 99.7% (95% CI, 96.7–100) and pooled specificity 10.8% (95% CI, 6.7–16.9). The efficiency (proportion of patients stratified into the ‘PE unlikely’ group) was 8.4% (95% CI, 4.1–16.5), and the failure rate (proportion of low likelihood patients who were diagnosed with PE during follow-up) was 0.1% (95% CI, 0–5.3). Conclusion In symptomatic hospitalized patients, use of the Wells rule plus D-dimer to rule out PE is safe, but allows very few patients to forgo imaging.

Background: Undernutrition among mothers and children is currently a major public health and development concern in Bangladesh. In literature relating to nutritional determinants, of particular interest is the geography, as regions with poor nutrition tend to pull down the overall nutritional status of the country. As such, reducing the regional gap can alone reduce overall undernutrition significantly, especially when regional gaps are high. The aim of this study is, therefore, to assess the magnitude of inequalities in undernutrition in children aged under 5 years in Bangladesh and their mothers, and relate this to the administrative divisions of the country. Methods: The Bangladesh Demographic and Health Surveys (1996–1997, 1999–2000, 2004 and 2007) were the sources of data, and a total of 16 278 mother–child pairs whose records were complete for the required individual and household-level variables were included in the analysis. Maternal nutritional status was measured by the body mass index (BMI). Weight-for-age, height-for-age and weight-for-height z-scores were calculated by use of the World Health Organization (WHO) Child Growth Standards to assess the nutritional status of children aged under 5 years. General linear model, sequential linear and multinomial logistic regression analyses were done to assess the inequalities in maternal and child nutritional status among the six administrative divisions of Bangladesh. Socioeconomic variables that were controlled for were residency, education and occupation of the mothers and their husbands, house type and possession score in the household. Results: Maternal BMI and prevalence of underweight, stunting and wasting in children aged under 5 years were found to vary significantly according to administrative division. Of the six divisions, Sylhet was found to have highest prevalence of undernourished mothers and children. The trends from 1996 to 2007 also established Sylhet as the poorest-performing region overall. Conclusion: The Sylhet administrative division needs specially focused attention from policy-makers if the overall performance of the health, nutrition and population sector is to reach the targets set by the country.

Primordial germ cells (PGCs) appear during early embryogenesis and differentiate into gametes through oogenesis or spermatogenesis. Teleost PGCs can be visualized by injecting RNA transcribed from the fusion product of a fluorescent protein gene attached to the 3' untranslated region (3'UTR) of zebrafish nanos3 (zf-nos3). Although this method has been widely applied to teleost PGCs, the visualization of PGCs in pelagic species that have eggs with a hard chorion is more problematic due to the technical difficulty of microinjection into their eggs. In this study, we developed a reliable method for microinjection of fertilized eggs in a pelagic species, the barfin flounder. Using a microneedle with a constriction "brake", we were able to introduce gfp-nos3 3'UTR mRNA into embryos and to determine the origin and migration route of PGCs. We also isolated the barfin flounder nos3 (bf-nos3) gene to compare its 3'UTR sequence with that of zebrafish. The 3'UTR of the bf-nos3 sequence was longer than that of zf-nos3. However, PGCs were also visualized after injection of gfp-bf-nos3 3'UTR mRNA both in zebrafish and barfin flounder. These results suggest that the function of nos3 is conserved between these species regardless of the sequence differences. The method developed here for labeling PGCs with gfp-nos3 mRNA will provide a means to study PGC development in the embryos of a wide range of marine fish species.

The presence of germ cells in the early gonad is important for sexual fate determination and gonadal development in vertebrates. Recent studies in zebrafish and medaka have shown that a lack of germ cells in the early gonad induces sex reversal in favor of a male phenotype. However, it is uncertain whether the gonadal somatic cells or the germ cells are predominant in determining gonadal fate in other vertebrate. Here, we investigated the role of germ cells in gonadal differentiation in goldfish, a gonochoristic species that possesses an XX–XY genetic sex determination system. The primordial germ cells (PGCs) of the fish were eliminated during embryogenesis by injection of a morpholino oligonucleotide against the dead end gene. Fish without germ cells showed two types of gonadal morphology: one with an ovarian cavity; the other with seminiferous tubules. Next, we tested whether function could be restored to these empty gonads by transplantation of a single PGC into each embryo, and also determined the gonadal sex of the resulting germline chimeras. Transplantation of a single GFP-labeled PGC successfully produced a germline chimera in 42.7% of the embryos. Some of the adult germline chimeras had a developed gonad on one side that contained donor derived germ cells, while the contralateral gonad lacked any early germ cell stages. Female germline chimeras possessed a normal ovary and a germ-cell free ovary-like structure on the contralateral side; this structure was similar to those seen in female morphants. Male germline chimeras possessed a testis and a contralateral empty testis that contained some sperm in the tubular lumens. Analysis of aromatase, foxl2 and amh expression in gonads of morphants and germline chimeras suggested that somatic transdifferentiation did not occur. The offspring of fertile germline chimeras all had the donor-derived phenotype, indicating that germline replacement had occurred and that the transplanted PGC had rescued both female and male gonadal function. These findings suggest that the absence of germ cells did not affect the pathway for ovary or testis development and that phenotypic sex in goldfish is determined by somatic cells under genetic sex control rather than an interaction between the germ cells and somatic cells.

The feasibility of cryopreserving common carp (Cyprinus carpio) primordial germ cells (PGC) by vitrification of whole embryos at the 22- to 28-somite stage was investigated. Green fluorescent protein (GFP)-labeled PGC were cooled rapidly using liquid nitrogen after exposure to a pretreatment solution containing 1.5 M cryoprotectant (ethylene glycol or dimethyl sulfoxide, 30 or 50 min) and a vitrification solution containing 3 M cryoprotectant and 0.5 M sucrose (5, 10, 20, or 30 min). Embryonic cells that were pretreated for 30 min and vitrified for 20 min with ethylene glycol had the greatest rate of survival of embryonic cells (68.6%; P < 0.01), an optimal highest percentage of viable PGC (73.8 to 74.9%; P < 0.05), and no evidence of ice formation after thawing. The vitrified/thawed PGC were transplanted into blastula-stage embryos from goldfish (Carassius auratus). The PGC maintained their motility and moved to the gonadal ridge of the host embryo. Thus, the combination of vitrification and transplantation to produce germ-line chimeras is a powerful tool for the artificial production of next-generation offspring.

Primordial germ cells (PGCs) are the only cells in developing embryos with the potential to transmit genetic information to the next generation. We previously visualized the PGCs of several teleostean embryos by injecting RNA synthesized from constructs encoding green fluorescent protein (GFP) fused to the 3'UTR of the zebrafish ( Danio rerio ) nanos 1 gene ( nos 1). However, this technique was not always suitable for visualizing PGCs in embryos from all teleost species. In this study, we compared the visualization of PGCs in common carp ( Cyprinus carpio ) embryos using two artificial constructs containing GFP fused to the 3'UTR of nanos from either common carp or zebrafish. Visualization was better using GFP fused to the 3'UTR of the nanos gene from common carp, compared with that from zebrafish. The visualized PGCs successfully migrated toward the gonadal ridge after transplantation into goldfish host embryos, suggesting that they maintained normal migratory motility. These techniques could be useful for the production of inter-specific germline chimeras using common carp donor PGCs.

Germ cell-deficient fish usually develop as phenotypic males. Thus, the presence of germ cells is generally considered to be essential for female gonadal differentiation or the maintenance of ovarian structure. However, little is known of the role of germ cells in the determination of the sexual fate of gonadal somatic cells. We have established an inducible germ cell deficiency system in the loach ( Misgurnus anguillicaudatus , Cypriniformes: Cobitidae), a small freshwater fish, using knockdown of the dead end gene with a morpholino antisense oligonucleotide. Interestingly, loach lacking germ cells could develop as either phenotypic males or females, as characterized morphologically by the presence or absence of bony plates in the pectoral fins, respectively. The phenotypic males and females had testicular and ovarian structures, respectively, but lacked germ cells. Gene expression patterns in these male and female germ cell-deficient gonads were essentially the same as those in gonads of normal fish. Our observations indicate that sexually dimorphic gonads can develop in germ cell-deficient loach. In contrast to the situation in other model fish species, the gonadal somatic cells in phenotypic females autonomously differentiated into ovarian tissues and also played a role in the maintenance of gonadal structure. On the basis of our observations, we propose two possible models to explain the role of germ cells in sex determination in fish.

Summary Germ-line chimeras provide a valuable approach to surrogate propagation in poultry farming and aquaculture. The appropriate combinations of donor germ cells and host individuals enable the production of eggs and sperm and of offspring from strains with a high commercial value or valuable genotype via an easily reared species or strain. Primordial germ cells (PGCs) are one of the candidate cell types that can be used in the production of germ-line chimeras. The model teleost species, zebrafish, has been used to study specification, differentiation and migration of PGCs. However, it is important that the properties of PGCs are also determined for fish species used in aquaculture. Recent technical developments together with improvements in embryo manipulation techniques have enabled analyses of the differentiation of PGCs and their migration to the genital ridge. Here, we report the characteristic properties of PGCs from several teleost species and investigate their behavior in germ-line chimeras using fluorescently-tagged PGCs.

ObjectiveTo determine how much of the variation in nutritional status of Bangladeshi children under 5 years old can be attributed to the socio-economic status of the family.DesignNutritional status used reference Z-scores of weight-for-age (WAZ), height-for-age (HAZ) and weight-for-height (WHZ). A ‘possession score’ was generated based on ownership of a radio, television, bicycle, motorcycle and telephone, and the availability of electricity, with categories of 0 to 4+ possessions. A five-point (quintile) ‘poverty index’ was created using principal component analysis.SettingThe Bangladesh Demographic and Health Survey 2004 was the source of data.SubjectsA sample of 4891 children aged ResultsSome 57·8 % of the sample was either stunted, wasted or underweight (7·7 % were stunted, wasted and underweight). Of those stunted (48·4 %), 25·7 % were also underweight. Underweight and wasting prevalences were 40·7 % and 14·3 %, respectively. Mean WAZ, HAZ and WHZ did not differ by sex. Children of mothers with no education or no possessions were, on average, about 1 sd more underweight and stunted than those with higher educated mothers or with 4+ possessions. The possession score provided much greater discrimination of undernutrition than the poverty index. Nearly 50 % of children from households with no possessions were stunted, wasted or underweight (only 27 % in the poorest quintile), compared with only 3–6 % of children from households with 4+ possessions (over 13 % in the richest quintile).ConclusionsMaternal education and possession score were the main predictors of a child’s nutritional status. Possession score was a much better indicator of undernutrition than the poverty index.

Primordial germ cells (PGCs) are the only cells in developing embryos with the potential to transmit genetic information to the next generation. In our previous study, a single PGC transplanted into a host differentiated into fertile gametes and produced germ-line chimeras of cyprinid fish, including zebrafish. In this study, we aimed to induce germ-line chimeras by transplanting donor PGCs from various sources (normal embryos at different stages, dissociated blastomeres, embryoids, or embryoids cryopreserved by vitrification) into host blastulae, and compare the migration rates of the PGCs towards the gonadal ridge. Isolated, cultured blastomeres not subject to mesodermal induction were able to differentiate into PGCs that retained their motility. Moreover, these PGCs successfully migrated towards the gonadal ridge of the host and formed viable gametes. Motility depended on developmental stage and culture duration: PGCs obtained at earlier developmental stages and with shorter cultivation periods showed an increased rate of migration to the gonadal ridge. Offspring were obtained from natural spawning between normal females and chimeric males. These results provide the basis for new methods of gene preservation in zebrafish.