Leveraging Integrated RNA Sequencing to Decipher Adrenomedullin's Protective Mechanisms in Experimental Bronchopulmonary Dysplasia.

Bronchopulmonary dysplasia (BPD) is a chronic lung disease commonly affecting premature infants, with limited therapeutic options and increased long-term consequences. Adrenomedullin (Adm), a proangiogenic peptide hormone, has been found to protect rodents against experimental BPD. This study aims to elucidate the molecular and cellular mechanisms through which Adm influences BPD pathogenesis using a lipopolysaccharide (LPS)-induced model of experimental BPD in mice. Bulk RNA sequencing of Adm-sufficient (wild-type or Adm+/+) and Adm-haplodeficient (Adm+/-) mice lungs, integrated with single-cell RNA sequencing data, revealed distinct gene expression patterns and cell type alterations associated with Adm deficiency and LPS exposure. Notably, computational integration with cell atlas data revealed that Adm-haplodeficient mouse lungs exhibited gene expression signatures characteristic of increased inflammation, natural killer (NK) cell frequency, and decreased endothelial cell and type II pneumocyte frequency. Furthermore, in silico human BPD patient data analysis supported our cell type frequency finding, highlighting elevated NK cells in BPD infants. These results underscore the protective role of Adm in experimental BPD and emphasize that it is a potential therapeutic target for BPD infants with an inflammatory phenotype.

Amphiregulin Exerts Proangiogenic Effects in Developing Murine Lungs.

Interrupted lung angiogenesis is a hallmark of bronchopulmonary dysplasia (BPD); however, druggable targets that can rescue this phenotype remain elusive. Thus, our investigation focused on amphiregulin (Areg), a growth factor that mediates cellular proliferation, differentiation, migration, survival, and repair. While Areg promotes lung branching morphogenesis, its effect on endothelial cell (EC) homeostasis in developing lungs is understudied. Therefore, we hypothesized that Areg promotes the proangiogenic ability of the ECs in developing murine lungs exposed to hyperoxia. Lung tissues were harvested from neonatal mice exposed to normoxia or hyperoxia to determine Areg expression. Next, we performed genetic loss-of-function and pharmacological gain-of-function studies in normoxia- and hyperoxia-exposed fetal murine lung ECs. Hyperoxia increased Areg mRNA levels and Areg+ cells in whole lungs. While Areg expression was increased in lung ECs exposed to hyperoxia, the expression of its signaling receptor, epidermal growth factor receptor, was decreased, indicating that hyperoxia reduces Areg signaling in lung ECs. Areg deficiency potentiated hyperoxia-mediated anti-angiogenic effects. In contrast, Areg treatment increased extracellular signal-regulated kinase activation and exerted proangiogenic effects. In conclusion, Areg promotes EC tubule formation in developing murine lungs exposed to hyperoxia.

Hyperoxia Disrupts Lung Lymphatic Homeostasis in Neonatal Mice.

Inflammation causes bronchopulmonary dysplasia (BPD), a common lung disease of preterm infants. One reason this disease lacks specific therapies is the paucity of information on the mechanisms regulating inflammation in developing lungs. We address this gap by characterizing the lymphatic phenotype in an experimental BPD model because lymphatics are major regulators of immune homeostasis. We hypothesized that hyperoxia (HO), a major risk factor for experimental and human BPD, disrupts lymphatic endothelial homeostasis using neonatal mice and human dermal lymphatic endothelial cells (HDLECs). Exposure to 70% O2 for 24-72 h decreased the expression of prospero homeobox 1 (Prox1) and vascular endothelial growth factor c (Vegf-c) and increased the expression of heme oxygenase 1 and NAD(P)H dehydrogenase [quinone]1 in HDLECs, and reduced their tubule formation ability. Next, we determined Prox1 and Vegf-c mRNA levels on postnatal days (P) 7 and 14 in neonatal murine lungs. The mRNA levels of these genes increased from P7 to P14, and 70% O2 exposure for 14 d (HO) attenuated this physiological increase in pro-lymphatic factors. Further, HO exposure decreased VEGFR3+ and podoplanin+ lymphatic vessel density and lymphatic function in neonatal murine lungs. Collectively, our results validate the hypothesis that HO disrupts lymphatic endothelial homeostasis.

Birth registration in Nepal: An assessment of progress based on two national surveys.

Birth registration, an essential component of the civil registration system, is expected to be complete and universal. This study assesses the progress made in recent years and identifies gaps in birth registration in Nepal. Data from the Multiple Indicator Cluster Surveys undertaken in 2014 and 2019 are used for the analysis. The two surveys included a total of 12,007 children under five years of age living with their mothers at the time of the surveys. The survey respondents were 11,821 mothers and 186 caretakers (in the case of those without mothers) of the children. The variations in the proportion of births registered among various subgroups of the children are assessed by performing bivariate analysis and binary logistic regression. Birth registration increased considerably, from 58% (95% CI: 57-59%) in 2014 to 77% (95% CI: 76-78%) in 2019. Several of the disparities between and among the various population subgroups that were evident in the 2014 survey had been considerably reduced or eliminated by 2019. The disparities in registration between boys and girls attenuated over time. Although birth registration increased for all children (ages 0-59 months old), infants still had comparatively lower levels of registration. The relatively disadvantaged provinces showed significant progress between the two survey periods. Considerable and significant progress has been made in birth registration in recent years. However, achieving universal and complete birth registration would require sustaining recent achievements and applying proven strategic interventions to ensure the inclusion of the unregistered births.

Extracellular Signal-Regulated Kinase 1 Alone Is Dispensable for Hyperoxia-Mediated Alveolar and Pulmonary Vascular Simplification in Neonatal Mice.

Bronchopulmonary dysplasia (BPD) is a morbid lung disease distinguished by lung alveolar and vascular simplification. Hyperoxia, an important BPD causative factor, increases extracellular signal-regulated kinases (ERK)-1/2 expression, whereas decreased lung endothelial cell ERK2 expression reduces angiogenesis and potentiates hyperoxia-mediated BPD in mice. However, ERK1's role in experimental BPD is unclear. Thus, we hypothesized that hyperoxia-induced experimental BPD would be more severe in global ERK1-knockout (ERK1-/-) mice than their wild-type (ERK1+/+ mice) littermates. We determined the extent of lung development, ERK1/2 expression, inflammation, and oxidative stress in ERK1-/- and ERK1+/+ mice exposed to normoxia (FiO2 21%) or hyperoxia (FiO2 70%). We also quantified the extent of angiogenesis and hydrogen peroxide (H2O2) production in hyperoxia-exposed neonatal human pulmonary microvascular endothelial cells (HPMECs) with normal and decreased ERK1 signaling. Compared with ERK1+/+ mice, ERK1-/- mice displayed increased pulmonary ERK2 activation upon hyperoxia exposure. However, the extent of hyperoxia-induced inflammation, oxidative stress, and interrupted lung development was similar in ERK1-/- and ERK1+/+ mice. ERK1 knockdown in HPMECs increased ERK2 activation at baseline, but did not affect in vitro angiogenesis and hyperoxia-induced H2O2 production. Thus, we conclude ERK1 is dispensable for hyperoxia-induced experimental BPD due to compensatory ERK2 activation.

Fire and forage quality: Postfire regrowth quality and pyric herbivory in subtropical grasslands of Nepal.

Fire is rampant throughout subtropical South and Southeast Asian grasslands. However, very little is known about the role of fire and pyric herbivory on the functioning of highly productive subtropical monsoon grasslands lying within the Cwa climatic region. We assessed the temporal effect of fire on postfire regrowth quality and associated pyric-herbivory in the subtropical monsoon grasslands of Bardia National Park, Nepal. Every year, grasslands are burned as a management intervention in the park, especially between March and May. Within a week after fire, at the end of March 2020, we established 60 m × 60 m plots within patches of burned grassland in the core area of the Park. We collected grass samples from the plots and determined physical and chemical properties of the vegetation at regular 30-day intervals from April to July 2020, starting from 30 days after fire to assess postfire regrowth forage quality. We counted pellet groups of cervids that are abundant in the area for the same four months from 2 m × 2 m quadrats that were permanently marked with pegs along the diagonal of each 60 m × 60 m plot to estimate intensity of use by deer to the progression of postfire regrowth. We observed strong and significant reductions in crude protein (mean value 9.1 to 4.1 [55% decrease]) and phosphorus (mean value 0.2 to 0.11 [45% decrease]) in forage collected during different time intervals, that is, from 30 days to 120 days after fire. Deer utilized the burned areas extensively for a short period, that is, up to two months after fire when the burned areas contained short grasses with a higher level of crude protein and phosphorus. The level of use of postfire regrowth by chital (Axis axis) differed significantly over time since fire, with higher intensity of use at 30 days after fire. The level of use of postfire regrowth by swamp deer (Rucervus duvaucelii) did not differ significantly until 90 days after fire, however, decreased significantly after 90 days since fire. Large-scale single event fires, thus, may not fulfil nutritional requirements of all species in the deer assemblage in these subtropical monsoon grasslands. This is likely because the nutritional requirements of herbivores differ due to differences in body size and physiological needs-maintenance, reproduction, and lactation. We recommend a spatiotemporal manipulation of fire to reinforce grazing feedback and to yield forage of high quality for the longest possible period for a sustainable high number of deer to maintain a viable tiger population within the park.

dbDNA™: An advanced platform for genetic medicines.

The supply of plasmid DNA has become a major bottle neck in the ever-expanding genetic medicine sector. Therefore, the development of new, scalable, faster DNA production technologies is vital for this sector going forward.

Leopard (Panthera pardus) occupancy in the Chure range of Nepal.

Conservation of large carnivores such as leopards requires large and interconnected habitats. Despite the wide geographic range of the leopard globally, only 17% of their habitat is within protected areas. Leopards are widely distributed in Nepal, but their population status and occupancy are poorly understood. We carried out the sign-based leopard occupancy survey across the entire Chure range (~19,000 km2) to understand the habitat occupancy along with the covariates affecting their occupancy. Leopard signs were obtained from in 70 out of 223 grids surveyed, with a naïve leopard occupancy of 0.31. The model-averaged leopard occupancy was estimated to be 0.5732 (SE 0.0082) with a replication-level detection probability of 0.2554 (SE 0.1142). The top model shows the additive effect of wild boar, ruggedness, presence of livestock, and human population density positively affecting the leopard occupancy. The detection probability of leopard was higher outside the protected areas, less in the high NDVI (normalized difference vegetation index) areas, and higher in the areas with livestock presence. The presence of wild boar was strong predictor of leopard occupancy followed by the presence of livestock, ruggedness, and human population density. Leopard occupancy was higher in west Chure (0.70 ± SE 0.047) having five protected areas compared with east Chure (0.46 ± SE 0.043) with no protected areas. Protected areas and prey species had positive influence on leopard occupancy in west Chure range. Similarly in the east Chure, the leopard occupancy increased with prey, NDVI, and terrain ruggedness. Enhanced law enforcement and mass awareness activities are necessary to reduce poaching/killing of wild ungulates and leopards in the Chure range to increase leopard occupancy. In addition, maintaining the sufficient natural prey base can contribute to minimize the livestock depredation and hence decrease the human-leopard conflict in the Chure range.

Maternal Mortality Levels and Trends in Nepal: A Brief Update.

Recently revised estimates of levels and trends of maternal mortality based on three sources are reviewed and trajectory for reaching the mortality reducing target by 2030 is assessed. According to the estimates provided by the UN Maternal Mortality Inter-agency Group (MMEIG), the maternal mortality ratio (MMR) started from a high of 901 in 1990, dropping to 258 in 2015, or a reduction of 71% over the 25 year period. Between 2000 and 2015, MMEIG estimates showed a reduction of 53%. The MMR is targeted to be reduced to at least to 70 by 2030. This implies a reduction of 73%, which is considerably higher than the reduction recorded during the earlier (2000-2015) period (which was 53%), or annualized rate of decline of 8.7% v 5.0%. Therefore, the future trajectory warrants undertaking strategic interventions more intensively than what may have been hitherto. Concurrently, more attention also needs to be given to strengthening the recording and reporting of maternal deaths. Keywords: levels and trends; maternal mortality; Nepal; projections.

Clinical Outcomes With the Use of Prophylactic Versus Therapeutic Anticoagulation in Coronavirus Disease 2019.

OBJECTIVES: To determine the impact of anticoagulation on inhospital mortality among coronavirus disease 2019-positive patients with the a priori hypothesis that there would be a lower risk of inhospital mortality with use of preemptive therapeutic over prophylactic dose enoxaparin or heparin. DESIGN SETTING: Retrospective cohort study from April 1, 2020, to April 25, 2020. The date of final follow-up was June 12, 2020 Two large, acute-care hospitals in Western Connecticut. PATIENTS: Five hundred and one inpatients were identified after discharge as 18 years or older and positive for severe acute respiratory syndrome coronavirus 2. The final sample size included 374 patients after applying exclusion criteria. Demographic variables were collected via hospital billing inquiries, whereas the clinical variables were abstracted from patients' medical records. EXPOSURE: Preemptive enoxaparin or heparin at a therapeutic or prophylactic dose. MAIN RESULTS: When comparing treatments through multivariable analysis, risk of inhospital mortality was 2.3 times greater in patients receiving preemptive therapeutic anticoagulation (95% CI = 1.0-4.9; p = 0.04). Additionally, the average treatment effects were higher (ß = 0.11, p = 0.01) in the therapeutic group. CONCLUSIONS: An increase in inhospital mortality was observed among patients on preemptive therapeutic anticoagulation. Thus, in the management of coronavirus disease 2019 and its complications, we recommend further research and cautious use of preemptive therapeutic over prophylactic anticoagulation.

Complementary Role of Public and Private Hospitals for Utilizing Outpatient Services in a Hill District in Nepal.

BACKGROUND: In Nepal, the private sector has prominently emerged as a provider of health services in recent years. The objective of this paper is to assess whether public and private hospitals are competing for patients with similar socioeconomic strata, or providing services to different segments of the patient population. METHODS: Data were collected prospectively from one public hospital and one private-for-profit hospital, both located in close proximity to one another in Tanahu district. A total of 384 and 389 patients presenting themselves for outpatient services available at the district public hospital and a private hospital, respectively, were systematically selected and interviewed using a survey form. The profiles of the patients were comparatively analyzed, and the reasons for using a particular hospital were assessed. Binary logistic regression was used for multivariate analysis. RESULTS: Compared to the patients using the public hospital, patients at the private hospital were younger, possessed a higher level of education, represented indigenous and disadvantaged ethnic groups, and belonged to business or agricultural occupations. The four prominent reasons for using the private hospital were: positive perception/prior experience, followed by recommendation/word-of-mouth, timely availability of services, and trustworthiness. Among the public hospital patients, the prominent reasons were: low fee for services or having insurance, positive perception/prior experience, and trustworthiness. CONCLUSIONS: Public and private hospitals have played a complementary role in serving the health needs of different patient population segments in the study district.

A new wave in the quiet revolution in contraceptive use in Nepal: the rise of emergency contraception.

BACKGROUND: In Nepal, while the use of the emergency contraceptive pill (ECP) has been increasing rapidly in recent years, very little is known about the profile of ECP users. This study assesses the increasing role of ECP and the profile of ECP users in Nepal. Some policy and programmatic concerns are also addressed. METHODS: Survey interviews were undertaken involving 185 women who visited five metropolitan medical shops located in Pokhara seeking to purchase ECP for their own use. Descriptive statistics were calculated for data analysis. RESULTS: ECP is popular among young (<25 years old) and educated women. Also, nearly 70 % of the ECP users described their most recent sexual relationship as infrequent/casual. The overwhelming majority of users are aware that ECP is for emergency use only. Most ECP users are also aware of other options like condoms, the oral pill, and injectable contraceptives; and a considerable proportion of women using ECP had also used these methods before. CONCLUSION: ECP is filling an important and unique role in reducing unplanned or unwanted pregnancy, especially among young, educated women, and those with infrequent sexual relationships.

Adverse consequences of unintended pregnancy for maternal and child health in Nepal.

In Nepal, 26%-38% of recent births are estimated to be from unintended pregnancies, but little is known whether these pregnancies have adverse consequences for the health of the mother and child. Data from the 2011 Nepal Demographic and Health Survey are used to examine the hypothesis that unintended pregnancies are associated with negative health outcomes for both mothers and children. When the pregnancy was unintended (compared with when it was intended) mothers were more likely to receive inadequate prenatal care (odds ratio OR = 1.50; 95% confidence interval [CI] = 1.28-1.77). They were also more likely to opt for home births (OR = 1.30; 95% CI = 1.11-1.52). Likewise, the resultant newborns of unintended pregnancies were more likely to receive inadequate immunization (OR = 1.18; 95% CI = 1.00-1.40) and to remain stunted (OR = 1.25; 95% CI = 1.00-1.56). Findings suggest significant associations between unintended pregnancy and negative health outcomes for both mothers and children in Nepal.

Women's awareness of liberalization of abortion law and knowledge of place for obtaining services in Nepal.

In Nepal, following the liberalization of the abortion law, expansion and scaling up of services proceeded in parallel with efforts to create awareness of the legalization status of abortion and provide women with information about where services are available. This article assesses the effectiveness of these programmatic interventions in the early years of the country's abortion program. Data from a 2006 national survey are analyzed with 2 outcome measures-awareness of the legal status of abortion and knowledge of places to obtain abortion services among women ages 15 to 44 years. The variations in the outcomes are analyzed by ecological-development subregion, residence, education, household wealth quintile, age, and number of living children. Bivariate and multivariate logistic regression techniques are used. Overall 32.3% (95% confidence interval = 31.4% to 33.2%) of the respondents were aware of the legal status of abortion and 56.5% (95% confidence interval = 55.5% to 57.4%) knew of a place where they could obtain an abortion. Both outcome measures showed considerable variations by the covariates. Women with secondary or higher level of education had the highest odds ratio of being aware of the law and having knowledge of a source for abortion services. Ecological-development subregions showed the second highest levels of odds ratios. Significant disparities among the population subgroups existed in the diffusion of awareness of the legal status of abortion and having knowledge of a place for abortion services in Nepal. The results point to which population subgroups to focus on and also serve as a baseline for assessing future progress in the diffusion process.

Contraceptive behavior among women after abortion in Nepal.

OBJECTIVE: To investigate the timing of contraceptive use and estimate the discontinuation rates of temporary methods among women after abortion, or a live birth or stillbirth. METHODS: A sample of married women with detailed pregnancy and contraceptive histories was analyzed in a population-based cross-sectional study based on calendar data extracted from the 2011 Nepal Demographic and Health Survey. Kaplan-Meier cumulative and discrete-time hazard models were used to estimate the timing of contraceptive use and discontinuation rates among users of temporary methods. RESULTS: The final analysis sample included 3190 women. Of 684 women who had had an abortion, 298 (43.6%) had not initiated any contraceptive use in the 12 months afterwards. Women initiated contraceptive use significantly earlier after abortion (hazard ratio [HR] 2.25; 95% CI, 1.96-2.59; P<0.001). The rate of discontinuation among contraceptive users was significantly higher in the postabortion group (HR 1.32; 95% CI, 1.05-1.65; P<0.05). Women who were educated, wealthier, had used contraceptives before the index pregnancy, had two sons and had autonomy initiated contraceptive use significantly earlier in the post-abortion period than their counterparts. CONCLUSION: Postabortion contraceptive use is low in Nepal. Postabortion family-planning counseling and related services should be strengthened with systematic monitoring and follow-up interventions.

Women's knowledge of abortion law and availability of services in Nepal.

This paper assesses women's awareness of the liberalization of abortion law and their knowledge of a place for obtaining abortion services in Nepal. The data are from the 2011 Nepal Demographic and Health Survey. The results are compared with data from a similar survey conducted in 2006. Variations in the two measures among several population sub-groups are analysed by performing logistic regression. Among women aged 15-44, 38.7% (CI: 37.8, 39.6) were aware of the legal status of abortion and 59.8% (CI: 58.9, 60.7) knew of a place to have an abortion. The percentages of both measures varied considerably by various population sub-groups. Over a 5-year period, knowledge of the legality of abortion increased by 6.4 percentage points, and awareness of service delivery sites increased by 3.3 percentage points. The increases in both measures were, however, largely limited to higher wealth quintiles and those with higher educational attainment. The results suggest the need to intensify efforts to educate women in Nepal, particularly the most disadvantaged women, about abortion law, including the conditions under which abortion is permitted, and where to access safe abortion services.

Abortion clients of a public-sector clinic and a non-governmental organization clinic in Nepal.

This paper investigates similarities and differences between abortion clients of a public-sector clinic and a non-governmental organization (NGO) clinic in Nepal. In 2010, a survey of 1,172 women was conducted in two highly-attended abortion clinics in Kathmandu-one public-sector clinic and another operated by an NGO. Data on the sociodemographic characteristics of clients, their fertility preferences, and use of contraceptives were analyzed. Similarities and differences between the two groups of clients were examined by either chi-square or t-test. The clients of the two clinics were similar with respect to age (27.3+/-5.7 years), education (26.5% had no education), and number of living children (1.88+/-1.08). They differed with regard to contraceptive practice, the circumstances resulting in unintended pregnancy, and future fertility preferences. Just over 50% clients of the public and 35% clients of the NGO clinic reported use of contraceptives surrounding the time of unintended pregnancy. The groups also differed in the contraceptive methods used and in reasons for not using any method. The NGO clinic contributed principally to expanding the availability of and access to abortion services.

Risk factors for repeat abortion in Nepal.

OBJECTIVE: To examine the incidence of and risk factors for repeat abortion in Nepal. METHODS: Data were analyzed from a survey of 1172 women who had surgical abortions between December 2009 and March 2010 in 2 clinics in Kathmandu, Nepal. Bivariate and multivariate logistic regressions were performed to estimate odds ratios for the risk factors. RESULTS: Among the respondents, 32.3% (95% confidence interval, 29.6-34.9) had repeat abortions. This incidence rose sharply with age and parity, and was higher among those with no intention of having a future child, those attaining primary or secondary level education, and those attending the non-governmental sector clinic. Women with repeat abortion were similar to those with 1 abortion in terms of contraceptive practice. Among women not using contraceptives at the time of the unintended pregnancy, the 3 most commonly cited reasons were ill health, non-compliance with the method intended for use, and dislike of the method. Women with repeat abortion showed a pattern of contraceptive acceptance immediately after the procedure similar to that of women who had 1 abortion. CONCLUSION: Repeat abortion is emerging as a major public health issue in Nepal, with implications for counseling and provision of abortion, and for family planning services.

Safe abortion services in Nepal: initial years of availability and utilization.

INTRODUCTION: Following the liberalization of the very strict Nepalese abortion law in 2002, the first services for safe induced abortion were introduced in 2004 at the nation's largest women's hospital. This paper examines the client profile, the context of demand for services, affordability and satisfaction with services. DATA AND METHODS: Data for the analysis came from a survey of women who presented themselves at the hospital for induced abortion services and subsequently received the services. RESULTS: Based on a survey of 672 clients, the median age was 26, and most women were married with an average of two living children. The majority reported being impregnated by the husband. Nearly three out of five gave their primary reason for termination as already having the number of children desired; another 42% cited finances. About two-thirds made the decision to abort jointly with the male partner. Most were satisfied with the services received and expenses incurred. About two-fifths reported having used a modern contraceptive method at the time the unwanted pregnancy occurred, while 22.6% reported practising either the safe-period or withdrawal methods. CONCLUSION: The clinic has provided affordable, quality abortion services to women in need. Findings also suggest that many areas need services strengthened, including the continued role of the family planning program in preventing unintended pregnancies.