Uporaba mikrosatelitskih biljega za utvrđivanje genetske varijabilnosti kod dugotrajno selekcioniranih linija crvenih rodajland kokoši

Genetic improvement needs the existence and precise estimation of genetic variability, and microsatellites are a molecular marker of choice for its assessment. Rhode Island Red (RIR) is a brown-egger chicken breed. A RIR population, selected on the basis of 40-week part-period egg production for 30 generations, maintained as a closed flock to develop multi-colored strains for backyard farming, were studied to determine genetic variability and heterozygosity using microsatellite markers. Genomic DNA from 76 randomly selected birds was analyzed at 10 microsatellite loci, alleles were separated on 3.4% MetaPhor Agarose, sized using Quantity-One software and analyzed by POPGENE v1.32. Nine loci demonstrated polymorphism resolving 30 alleles, the average number of alleles/ locus being 3.00 ± 1.41. Allele numbers varied from two to six and size ranged from 102 to 320bp. The average polymorphic information content and Nei’s heterozygosity were 0.3134 ± 0.064 and 0.4119 ± 0.2575, respectively, suggesting these loci to be moderately polymorphic and informative. The observed (Na) and effective number (Ne) of alleles and Shannon’s index averaged 3.0000 ± 1.4142, 2.0324 ± 0.9416 and 0.7342 ± 0.4649, respectively. Ne was less than Na at most loci, indicating the prevalence of heterozygosity at these loci. Mean expected heterozygosity was higher than mean observed heterozygosity at most loci, indicating that the population was not in Hardy-Weinberg equilibrium but was under the influence of some forces such as selection, etc. The same was also demonstrated by Chi-square and G-square tests. It may be inferred that long-term selection for 40-week part-period egg production has resulted in Hardy-Weinberg disequilibrium at the studied microsatellite loci, thereby suggesting a probable association between these microsatellite loci and layer economic traits in RIR chicken, and this might be useful in marker assisted selection for egg production in future.Genetsko poboljšanje traži postojanu i preciznu procjenu genetske varijabilnosti. S tom svrhom moguće je koristiti mikrosatelite kao molekularne biljege. Crveni rodajland je pasmina kokoši koja nese smeđa jaja, a ovo je istraživanje provedeno u populaciji koja je kroz 30 generacija selekcionirana na osnovi 40-tjedne proizvodnje jaja. Populacija je održavana kao zatvoreno jato namijenjeno za razvoj višebojnih linija koje bi se držale u dvorišnom tipu uzgoja. Cilj istraživanja bio je utvrditi genetsku varijabilnost i heterozigotnost primjenom mikrosatelitskih biljega. Genomska DNA od 76 slučajno odabranih kokoši analizirana je na 10 mikrosatelitskih lokusa. Aleli su razlučeni na 3,4 % MetaPhor agarozi, a njihova veličina i analiza određeni su pomoću Quantity-One i POPGENE v1.32 računalnih programa. Devet lokusa bilo je polimorfno pokazujući 30 alela, pri čemu je prosječni broj alela/lokusu iznosio 3,00 ± 1,41. Broj alela kretao se od 2 do 6, a veličina u rasponu 102 do 320 bp. Prosječna informativnost polimorfizma i Neieva heterozigotnost iznosili su 0,3134 ± 0,064 te 0,4119 ± 0,2575, što pokazuje da su promatrani lokusi umjereno polimorfni i informativni. Opaženi (Na) i efektivni broj (Ne) alela te Shannonov indeks imali su prosječne vrijednosti 3,0000 ± 1,4142, zatim 2,0324 ± 0,9416 i 0,7342 ± 0,4649. Ne je bio manji od Na na većini lokusa upujući na prevalenciju heterozigotnosti na tim lokusima. Na većini lokusa srednja očekivana heterozigotnost bila je veća u odnosu na srednju opaženu heterozigotnost, što je pokazalo da populacija nije bila u Hardy-Weinbergovoj ravnoteži već je bila pod utjecajem nekih sila kao npr. selekcije ili drugo. Isto je potvrđeno Hi-kvadrat i G-kvadrat testovima. Može se zaključiti da je dugotrajna selekcija s obzirom na 40-tjednu proizvodnju jaja rezultirala Hardy-Weinbergovom neravnotežom na istraženim mikrosatelitskim lokusima što upućuje na moguću povezanost između tih mikrosatelitskih lokusa i gospodarski važnih obilježja nesivosti kod kokoši crvenog rodajlanda. Navedeno bi moglo biti korisno u budućoj selekciji potpomognutoj s biljezima za proizvodnju jaja

Risk of Major Congenital Malformations and Exposure to Antiseizure Medication Monotherapy

Importance Women with epilepsy (WWE) require treatment with antiseizure medications (ASMs) during pregnancy, which may be associated with an increased risk of major congenital malformations (MCMs) in their offspring. Objective To investigate the prevalence of MCMs after prenatal exposure to 8 commonly used ASM monotherapies and changes in MCM prevalence over time. Design, Setting, and Participants This was a prospective, observational, longitudinal cohort study conducted from June 1999 to October 2022. Since 1999, physicians from more than 40 countries enrolled ASM-treated WWE before pregnancy outcome was known and followed up their offspring until 1 year after birth. Participants aged 14 to 55 years who were exposed to 8 of the most frequently used ASMs during pregnancy were included in this study. Data were analyzed from April to September 2023. Exposure Maternal use of ASMs at conception. Main Outcomes and Measures MCMs were assessed 1 year after birth by a committee blinded to type of exposure. Teratogenic outcomes across exposures were compared by random-effects logistic regression adjusting for potential confounders and prognostic factors. Results A total of 10 121 prospective pregnancies exposed to ASM monotherapy met eligibility criteria. Of those, 9840 were exposed to the 8 most frequently used ASMs. The 9840 pregnancies occurred in 8483 women (mean [range] age, 30.1 [14.1-55.2] years). MCMs occurred in 153 of 1549 pregnancies for valproate (9.9%; 95% CI, 8.5%-11.5%), 9 of 142 for phenytoin (6.3%; 95% CI, 3.4%-11.6%), 21 of 338 for phenobarbital (6.2%; 95% CI, 4.1%-9.3%), 121 of 2255 for carbamazepine (5.4%; 95% CI, 4.5%-6.4%), 10 of 204 for topiramate (4.9%; 95% CI, 2.7%-8.8%), 110 of 3584 for lamotrigine (3.1%; 95% CI, 2.5%-3.7%), 13 of 443 for oxcarbazepine (2.9%; 95% CI, 1.7%-5.0%), and 33 of 1325 for levetiracetam (2.5%; 95% CI, 1.8%-3.5%). For valproate, phenobarbital, and carbamazepine, there was a significant increase in the prevalence of MCMs associated with increasing dose of the ASM. Overall prevalence of MCMs decreased from 6.1% (153 of 2505) during the period 1998 to 2004 to 3.7% (76 of 2054) during the period 2015 to 2022. This decrease over time was significant in univariable logistic analysis but not after adjustment for changes in ASM exposure pattern. Conclusions and Relevance Of all ASMs with meaningful data, the lowest prevalence of MCMs was observed in offspring exposed to levetiracetam, oxcarbazepine, and lamotrigine. Prevalence of MCMs was higher with phenytoin, valproate, carbamazepine, and phenobarbital, and dose dependent for the latter 3 ASMs. The shift in exposure pattern over time with a declining exposure to valproate and carbamazepine and greater use of lamotrigine and levetiracetam was associated with a 39% decline in prevalence of MCMs, a finding that has major public health implications

Population-level risks of alcohol consumption by amount, geography, age, sex, and year: a systematic analysis for the Global Burden of Disease Study 2020

BACKGROUND: The health risks associated with moderate alcohol consumption continue to be debated. Small amounts of alcohol might lower the risk of some health outcomes but increase the risk of others, suggesting that the overall risk depends, in part, on background disease rates, which vary by region, age, sex, and year. METHODS: For this analysis, we constructed burden-weighted dose-response relative risk curves across 22 health outcomes to estimate the theoretical minimum risk exposure level (TMREL) and non-drinker equivalence (NDE), the consumption level at which the health risk is equivalent to that of a non-drinker, using disease rates from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2020 for 21 regions, including 204 countries and territories, by 5-year age group, sex, and year for individuals aged 15-95 years and older from 1990 to 2020. Based on the NDE, we quantified the population consuming harmful amounts of alcohol. FINDINGS: The burden-weighted relative risk curves for alcohol use varied by region and age. Among individuals aged 15-39 years in 2020, the TMREL varied between 0 (95% uncertainty interval 0-0) and 0·603 (0·400-1·00) standard drinks per day, and the NDE varied between 0·002 (0-0) and 1·75 (0·698-4·30) standard drinks per day. Among individuals aged 40 years and older, the burden-weighted relative risk curve was J-shaped for all regions, with a 2020 TMREL that ranged from 0·114 (0-0·403) to 1·87 (0·500-3·30) standard drinks per day and an NDE that ranged between 0·193 (0-0·900) and 6·94 (3·40-8·30) standard drinks per day. Among individuals consuming harmful amounts of alcohol in 2020, 59·1% (54·3-65·4) were aged 15-39 years and 76·9% (73·0-81·3) were male. INTERPRETATION: There is strong evidence to support recommendations on alcohol consumption varying by age and location. Stronger interventions, particularly those tailored towards younger individuals, are needed to reduce the substantial global health loss attributable to alcohol. FUNDING: Bill & Melinda Gates Foundation

Burnout among surgeons before and during the SARS-CoV-2 pandemic: an international survey

Background: SARS-CoV-2 pandemic has had many significant impacts within the surgical realm, and surgeons have been obligated to reconsider almost every aspect of daily clinical practice. Methods: This is a cross-sectional study reported in compliance with the CHERRIES guidelines and conducted through an online platform from June 14th to July 15th, 2020. The primary outcome was the burden of burnout during the pandemic indicated by the validated Shirom-Melamed Burnout Measure. Results: Nine hundred fifty-four surgeons completed the survey. The median length of practice was 10 years; 78.2% included were male with a median age of 37 years old, 39.5% were consultants, 68.9% were general surgeons, and 55.7% were affiliated with an academic institution. Overall, there was a significant increase in the mean burnout score during the pandemic; longer years of practice and older age were significantly associated with less burnout. There were significant reductions in the median number of outpatient visits, operated cases, on-call hours, emergency visits, and research work, so, 48.2% of respondents felt that the training resources were insufficient. The majority (81.3%) of respondents reported that their hospitals were included in the management of COVID-19, 66.5% felt their roles had been minimized; 41% were asked to assist in non-surgical medical practices, and 37.6% of respondents were included in COVID-19 management. Conclusions: There was a significant burnout among trainees. Almost all aspects of clinical and research activities were affected with a significant reduction in the volume of research, outpatient clinic visits, surgical procedures, on-call hours, and emergency cases hindering the training. Trial registration: The study was registered on clicaltrials.gov "NCT04433286" on 16/06/2020

The P323L substitution in the SARS-CoV-2 polymerase (NSP12) confers a selective advantage during infection

Background The mutational landscape of SARS-CoV-2 varies at the dominant viral genome sequence and minor genomic variant population. During the COVID-19 pandemic, an early substitution in the genome was the D614G change in the spike protein, associated with an increase in transmissibility. Genomes with D614G are accompanied by a P323L substitution in the viral polymerase (NSP12). However, P323L is not thought to be under strong selective pressure. Results Investigation of P323L/D614G substitutions in the population shows rapid emergence during the containment phase and early surge phase during the first wave. These substitutions emerge from minor genomic variants which become dominant viral genome sequence. This is investigated in vivo and in vitro using SARS-CoV-2 with P323 and D614 in the dominant genome sequence and L323 and G614 in the minor variant population. During infection, there is rapid selection of L323 into the dominant viral genome sequence but not G614. Reverse genetics is used to create two viruses (either P323 or L323) with the same genetic background. L323 shows greater abundance of viral RNA and proteins and a smaller plaque morphology than P323. Conclusions These data suggest that P323L is an important contribution in the emergence of variants with transmission advantages. Sequence analysis of viral populations suggests it may be possible to predict the emergence of a new variant based on tracking the frequency of minor variant genomes. The ability to predict an emerging variant of SARS-CoV-2 in the global landscape may aid in the evaluation of medical countermeasures and non-pharmaceutical interventions

Viral coinfections in hospitalized coronavirus disease 2019 patients recruited to the international severe acute respiratory and emerging infections consortium WHO clinical characterisation protocol UK study

Background We conducted this study to assess the prevalence of viral coinfection in a well characterized cohort of hospitalized coronavirus disease 2019 (COVID-19) patients and to investigate the impact of coinfection on disease severity. Methods Multiplex real-time polymerase chain reaction testing for endemic respiratory viruses was performed on upper respiratory tract samples from 1002 patients with COVID-19, aged <1 year to 102 years old, recruited to the International Severe Acute Respiratory and Emerging Infections Consortium WHO Clinical Characterisation Protocol UK study. Comprehensive demographic, clinical, and outcome data were collected prospectively up to 28 days post discharge. Results A coinfecting virus was detected in 20 (2.0%) participants. Multivariable analysis revealed no significant risk factors for coinfection, although this may be due to rarity of coinfection. Likewise, ordinal logistic regression analysis did not demonstrate a significant association between coinfection and increased disease severity. Conclusions Viral coinfection was rare among hospitalized COVID-19 patients in the United Kingdom during the first 18 months of the pandemic. With unbiased prospective sampling, we found no evidence of an association between viral coinfection and disease severity. Public health interventions disrupted normal seasonal transmission of respiratory viruses; relaxation of these measures mean it will be important to monitor the prevalence and impact of respiratory viral coinfections going forward

The Petrochemistry of Jake_M: A Martian Mugearite

“Jake_M,” the first rock analyzed by the Alpha Particle X-ray Spectrometer instrument on the Curiosity rover, differs substantially in chemical composition from other known martian igneous rocks: It is alkaline (&gt;15% normative nepheline) and relatively fractionated. Jake_M is compositionally similar to terrestrial mugearites, a rock type typically found at ocean islands and continental rifts. By analogy with these comparable terrestrial rocks, Jake_M could have been produced by extensive fractional crystallization of a primary alkaline or transitional magma at elevated pressure, with or without elevated water contents. The discovery of Jake_M suggests that alkaline magmas may be more abundant on Mars than on Earth and that Curiosity could encounter even more fractionated alkaline rocks (for example, phonolites and trachytes).</jats:p

Mapping inequalities in exclusive breastfeeding in low- and middle-income countries, 2000–2018

Exclusive breastfeeding (EBF)—giving infants only breast-milk for the first 6 months of life—is a component of optimal breastfeeding practices effective in preventing child morbidity and mortality. EBF practices are known to vary by population and comparable subnational estimates of prevalence and progress across low- and middle-income countries (LMICs) are required for planning policy and interventions. Here we present a geospatial analysis of EBF prevalence estimates from 2000 to 2018 across 94 LMICs mapped to policy-relevant administrative units (for example, districts), quantify subnational inequalities and their changes over time, and estimate probabilities of meeting the World Health Organization’s Global Nutrition Target (WHO GNT) of ≥70% EBF prevalence by 2030. While six LMICs are projected to meet the WHO GNT of ≥70% EBF prevalence at a national scale, only three are predicted to meet the target in all their district-level units by 2030