Generation of a homozygous mutant drug transporter (ABCB1) knockout line in the sea urchin Lytechinus pictus.

Sea urchins are premier model organisms for the study of early development. However, the lengthy generation times of commonly used species have precluded application of stable genetic approaches. Here, we use the painted sea urchin Lytechinus pictus to address this limitation and to generate a homozygous mutant sea urchin line. L. pictus has one of the shortest generation times of any currently used sea urchin. We leveraged this advantage to generate a knockout mutant of the sea urchin homolog of the drug transporter ABCB1, a major player in xenobiotic disposition for all animals. Using CRISPR/Cas9, we generated large fragment deletions of ABCB1 and used these readily detected deletions to rapidly genotype and breed mutant animals to homozygosity in the F2 generation. The knockout larvae are produced according to expected Mendelian distribution, exhibit reduced xenobiotic efflux activity and can be grown to maturity. This study represents a major step towards more sophisticated genetic manipulation of the sea urchin and the establishment of reproducible sea urchin animal resources.

90 versus 60 min of early skin-to-skin contact on exclusive breastfeeding rate in healthy infants' ≥35 weeks: A randomised controlled trial.

AIM: To compare the effect of 90 versus 60 min of early skin-to-skin contact (SSC) among vaginally born healthy infants ≥35 weeks of gestation on their exclusive breastfeeding rates and breastfeeding behaviour. METHODS: This parallel-group, open-label, randomised controlled trial enrolled healthy term and late preterm infants born vaginally. Infants in the intervention group received early SSC for 90 min compared to 60 min in the control group. The primary outcome was the proportion of infants on exclusive breastfeeding at 60 ± 12 h. RESULTS: One hundred ninety-eight mother-infant dyads were randomised (99 in each group). The infants in the 90-min SSC group were more likely to be exclusively breastfed at 60 ± 12 h as compared to the 60-min SSC group (RR, 95% CI-1.44, [1.15-1.79], p < 0.01). The modified infant breastfeeding assessment tool score at 60 ± 12 h was significantly higher in the 90-min SSC group (median [IQR]-9, [8, 10] versus 8 [7, 10], p = 0.03]. The proportion of infants on exclusive breastfeeding at 6, 10, and 14 weeks of age was also significantly higher in the 90-min SSC group (RR, 95% CI-1.39 [1.11-1.74], 1.36 [1.08-1.07], and 1.38 [1.08-1.75], respectively). CONCLUSION: Increasing the duration of early SSC showed a dose-response benefit on exclusive breastfeeding rates and breastfeeding behaviour. TRIAL REGISTRATION: CTRI/2018/09/015632, registered on 06/09/2018.

Cxcl9l and Cxcr3.2 regulate recruitment of osteoclast progenitors to bone matrix in a medaka osteoporosis model.

Bone homeostasis requires continuous remodeling of bone matrix to maintain structural integrity. This involves extensive communication between bone-forming osteoblasts and bone-resorbing osteoclasts to orchestrate balanced progenitor cell recruitment and activation. Only a few mediators controlling progenitor activation are known to date and have been targeted for intervention of bone disorders such as osteoporosis. To identify druggable pathways, we generated a medaka (Oryzias latipes) osteoporosis model, where inducible expression of receptor-activator of nuclear factor kappa-Β ligand (Rankl) leads to ectopic formation of osteoclasts and excessive bone resorption, which can be assessed by live imaging. Here we show that upon Rankl induction, osteoblast progenitors up-regulate expression of the chemokine ligand Cxcl9l. Ectopic expression of Cxcl9l recruits mpeg1-positive macrophages to bone matrix and triggers their differentiation into osteoclasts. We also demonstrate that the chemokine receptor Cxcr3.2 is expressed in a distinct subset of macrophages in the aorta-gonad-mesonephros (AGM). Live imaging revealed that upon Rankl induction, Cxcr3.2-positive macrophages get activated, migrate to bone matrix, and differentiate into osteoclasts. Importantly, mutations in cxcr3.2 prevent macrophage recruitment and osteoclast differentiation. Furthermore, Cxcr3.2 inhibition by the chemical antagonists AMG487 and NBI-74330 also reduced osteoclast recruitment and protected bone integrity against osteoporotic insult. Our data identify a mechanism for progenitor recruitment to bone resorption sites and Cxcl9l and Cxcr3.2 as potential druggable regulators of bone homeostasis and osteoporosis.

CRISPR/Cas9 mutagenesis reveals a role for ABCB1 in gut immune responses to Vibrio diazotrophicus in sea urchin larvae.

The ABC transporter ABCB1 plays an important role in the disposition of xenobiotics. Embryos of most species express high levels of this transporter in early development as a protective mechanism, but its native substrates are not known. Here, we used larvae of the sea urchin Strongylocentrotus purpuratus to characterize the early life expression and role of Sp-ABCB1a, a homolog of ABCB1. The results indicate that while Sp-ABCB1a is initially expressed ubiquitously, it becomes enriched in the developing gut. Using optimized CRISPR/Cas9 gene editing methods to achieve high editing efficiency in the F0 generation, we generated ABCB1a crispant embryos with significantly reduced transporter efflux activity. When infected with the opportunistic pathogen Vibrio diazotrophicus, Sp-ABCB1a crispant larvae demonstrated significantly stronger gut inflammation, immunocyte migration and cytokine Sp-IL-17 induction, as compared with infected control larvae. The results suggest an ancestral function of ABCB1 in host-microbial interactions, with implications for the survival of invertebrate larvae in the marine microbial environment.

SMN deficiency alters Nrxn2 expression and splicing in zebrafish and mouse models of spinal muscular atrophy.

Spinal muscular atrophy (SMA) is a progressive neurodegenerative disease affecting lower motor neurons. SMA is caused by mutations in the Survival Motor Neuron 1 (SMN1) gene, which result in reduced levels of functional SMN protein. Biochemical studies have linked the ubiquitously expressed SMN protein to the assembly of pre-mRNA processing U snRNPs, raising the possibility that aberrant splicing is a major defect in SMA. Accordingly, several transcripts affected upon SMN deficiency have been reported. A second function for SMN in axonal mRNA transport has also been proposed that may likewise contribute to the SMA phenotype. The underlying etiology of SMA, however, is still not fully understood. Here, we have used a combination of genomics and live Ca(2+) imaging to investigate the consequences of SMN deficiency in a zebrafish model of SMA. In a transcriptome analyses of SMN-deficient zebrafish, we identified neurexin2a (nrxn2a) as strongly down-regulated and displaying changes in alternative splicing patterns. Importantly, the knock-down of two distinct nrxn2a isoforms phenocopies SMN-deficient fish and results in a significant reduction of motor axon excitability. Interestingly, we observed altered expression and splicing of Nrxn2 also in motor neurons from the Smn(-/-);SMN2(+/+) mouse model of SMA, suggesting conservation of nrxn2 regulation by SMN in mammals. We propose that SMN deficiency affects splicing and abundance of nrxn2a. This may explain the pre-synaptic defects at neuromuscular endplates in SMA pathophysiology.

Quality of maternal and newborn health services and their impact on maternal-neonatal outcome at a primary health center.

Introduction: The utilization of the maternal and newborn health services has increased, but mere increase in utilization of services does not ensure that quality services are being provided. The aim of the study was to assess the quality of maternal and newborn services and their impact on maternal and neonatal outcome at a primary health center of Western Rajasthan in India. Materials and Methods: An exploratory study was undertaken at a conveniently selected primary health center providing 24-hour delivery services. Information regarding the availability of services was collected from the available medical officer in charge using an Indian Public Health Standards (IPHS) Proforma. Assessment of quality of services was performed by using WHO standards of care based on assessment of quality of maternal and newborn services tool by the perspectives of the provider as well as the mothers utilizing the services. 36 mothers who delivered at the selected PHC were interviewed. Results: All basic obstetric care services were available at the selected primary health centers including the 24 × 7 delivery services. The assessment of quality by provider's perspective revealed that the system of referral could be improved. Quality of maternal and newborn services assessment revealed that the practice of skin to skin contact between the mother and newborn just after the delivery was not being followed and few (30%) mothers informed that they could not start breastfeeding within 1 hours of birth. 47% mothers reported that they were not given the freedom to ask questions during delivery. Maternal and newborn outcome revealed that all mothers (100%) had a normal vaginal delivery, and 22% mothers had an episiotomy. All (100%) newborns cried immediately after birth, and average birthweight was 2.89 kg. Conclusion: PHCs are the first point of contact of mothers and healthcare delivery system. Assessment of quality of services is an important tool for quality assurance. Inclusion of evidence-based practices like skin-to-skin contact and early initiation of breastfeeding is important to improve the maternal and newborn well-being.

The Effect of Birthing Ball Exercises on Labor Pain and Labor Outcome Among Primigraviade Parturient Mothers at a Tertiary Care Hospital.

INTRODUCTION: Every woman has the right to respectful and empathetic care during childbirth that addresses her needs for pain management, and allows her the liberty to make it a memorable experience. This study aimed to assess the effect of birthing ball exercises on labor pain and labor outcome among primigravidae parturients at a tertiary care hospital. METHOD: A quasi-experimental design was used. A total of 60 primigravidae with 30 each in the control and experiment groups were selected by consecutive sampling. Primigravidae in the experiment group underwent two sessions of 20 minutes of birthing ball exercises at a subsequent gap of one hour during their active phase of labor (>4 cm cervical dilation). Primigravidae in the control group received routine standard care that included continuous observation and monitoring of vital signs and progress of labor. The visual analog scale (VAS) score was assessed in the transition phase (cervical dilation 8 cm to 10cm) and labor outcomes were assessed after delivery in both groups. RESULT: The experiment group had significantly better labor outcomes in terms of labor pain, cervical dilatation, and duration of labor compared to the primigravidae in the control group (p<0.05). In addition, the majority of mothers in the experiment group (86.7 %) underwent vaginal delivery with episiotomy compared to the control group (53.3%). Findings also revealed a statistically significant difference in the newborns of both groups regarding appearance, pulse, grimace, activity, and respiration (APGAR) score, crying immediately after birth, and admission to the neonatal intensive care unit (NICU) at p<0.05. CONCLUSION: There are a variety of discomforts that a woman experiences during labor. Reducing these discomforts is an important part of good nursing care. Non-pharmacologic methods like birthing ball exercises help decrease these discomforts by reducing labor pain and improving maternal and neonatal outcomes.

Effect of supported sitting position during second stage of labor on its outcome in primigravidae: A quasi-experimental study.

Background: The childbirth position has a significant influence on labor, maternal comfort, and neonatal outcome. In sitting position, there is a faster fetal descent with the effect of gravity. The information on this subject is relatively scant. Therefore, this study aimed to examine the effect of a supported sitting position during second stage of labor on its outcome in primigravidae. Materials and Methods: A quasi-experimental study with a post-test only control group design was used. 60 primigravidae were selected using total enumerative sampling. The labor outcome was assessed by self-structured maternal neonatal outcome checklist and socio-demographic proforma. Results: Statistically significant difference was observed on the mean duration of second stage of labor among primigravidae in control and experimental group (t = 5.87, P < 0.001) and also in the APGAR score of newborns (t = -3.98, P < 0.001). A statistical significant association of duration of second stage of labor with height and intensity of maternal work was also observed. Conclusions: A supported sitting position during labor was found to be effective in reducing duration of the second stage of labor. This can be used as a nursing intervention while providing care during labor especially at primary healthcare centers that can help in reducing the duration of second stage of labor.

Effectiveness of prenatal lactation counseling on breastfeeding practices, breast engorgement, and newborn feeding behavior among postnatal mothers at a teaching institution.

Context: Breastfeeding is the best way of providing nutrients to young infants for their healthy growth and development. World Health Organization (WHO) and UNICEF recommend that breastfeeding should be initiated as early as possible after delivery and exclusive breastfeeding should be continued till 6 months of age. The proper positioning of the baby and mother makes breastfeeding effective. Aim: The study aimed to determine the effect of lactation counseling on breastfeeding practices, breast engorgement, and newborn feeding behavior among postnatal mothers. Methods: A quasi-experiment posttest-only control group research design was used and the setting of the study was antenatal OPD and postnatal ward at a teaching institution. A sample of 60 primigravidae in the age group of 18-35 years with gestational age ≥ 36 weeks seeking antenatal care at antenatal OPD were selected by consecutive sampling technique. The experimental group received two lactation counseling sessions of 30 min each 1 week apart (in person/video call), whereas control group received routine care. Breastfeeding practices, breast engorgement, and newborn feeding behavior were assessed on the 3rd postnatal day using breastfeeding practices checklist, breast engorgement scale, and newborn feeding behavior assessment tool, respectively. Descriptive and inferential statistics were used to analyze the data. Result: There was significant improvement in breastfeeding practices (t = 7.18, P = 0.00), breast engorgement (t = 2.41, P = 0.01), and newborn feeding behavior (t = 5.24, P = 0.00) in the experimental group, which proves that the prenatal lactation counseling was effective in improving breastfeeding practices, newborn feeding behavior, and reducing breast engorgement. Conclusion: The study concluded that prenatal lactation counseling was effective in improving breastfeeding practices, breast engorgement, and newborn breastfeeding behavior among primigravidae. These findings suggest that if proper lactation counseling is provided to the mothers even at the primary health centers and periphery level it can contribute to improving the breastfeeding practices, newborn feeding behavior and can prevent breast engorgement.

Online learning in nursing students: Satisfaction and barriers.

BACKGROUND: The nationwide coronavirus (COVID-19) pandemic and ensuing lockdown has enforced institutions crosswise India to provisionally close to inhibit the spread of the virus and started online learning for students. To measure the level of satisfaction of nursing students with online learning and to identify the barriers which restrict to online learning. MATERIALS AND METHODS: The current study adopted quantitative research approach with an online survey research design and carried out during May-June 2020. Participants were selected through a web-based survey (Google form), in which 219 students enrolled. Self-structured questionnaire with the Likert scale was used to measure the level of satisfaction of nursing students with online learning and identify the barriers which restrict online learning. The descriptive and inferential statistics were used for the analysis in which 219 participants were enrolled in the study of data with IBM SPSS version 20. RESULTS: Majority of student's participants 148 (67.57%) were extremely satisfied with online learning. The findings suggest that the highest barriers which restrict to online learning among nursing students is low voice and language clarity (2.16 ± 0.593), physical health barriers such as eye strain (2.43 ± 0.613), reliability and connectivity problem (2.26 ± 0.534). Among all demographic data, age is significantly associated with the level of satisfaction of online learning. CONCLUSIONS: The study data indicated that maximum students were extremely satisfied the with online learning and among barriers which effect online learning is low voice and language clarity, reliability and connectivity problem, physical health barriers such as eye strain.

Stress and professional burnout among newly graduated dentists.

BACKGROUND: Dentists encounter numerous professional stressful situations, beginning from education to day-to-day practice. The resulting stress tends to have a negative impact on their personal as well as professional lives. OBJECTIVES: To measure daily burnout, and to investigate the extent of expectations from dental career and the feeling of being unqualified new dental practitioner. MATERIALS AND METHODS: A close-ended questionnaire, i.e., "the Copenhagen Burnout Inventory," was utilized for evaluation. A total of 121 dentists with an experience ranging between 6 months and 5 years were included. The period was considered initiating from graduation to dental practicing in urban or rural areas. Ninety-seven dentists replied with filled questionnaires (80.16%). The multivariant analysis was done using SPSS 11.0 ver. (Chicago, USA). RESULTS: Using measures analysis, the mean scores for dentists on the basis of age and sex (n = 97) were calculated. The factors most commonly considered responsible for professional burnout were emotional exhaustion (39.27%), frustrations (47.83%), feeling worn out at the end of the day (35.05%), feeling worn out at the end of the working day (46.80%), exhaustion in the morning at the thought of another day at work (35.05%), feeling that every working hour is tiring (46.80%), less energy and less time for family and friends (47.83%). The most common cause for stress was professional burnout that was recorded commonly in females in the age range of 26-28 years. CONCLUSIONS: Dentists are more prone for professional burnout, anxiety, and depression. The main reason for this is the nature of their practice and their personality traits, especially while pursuing dentistry as a carrier. Stress may lead to negative impact on dentists' personal as well as professional lives.