bluuebunny/crossref_metadata_2025 huggingface dataset:
American foulbrood in a honeybee colony: spore-symptom relationship and feedbacks between disease and colony development
Authors: Stephan Jörg G. , de Miranda Joachim R. , Forsgren Eva
Published: March 2020

Background

The most severe bacterial disease of honeybees is American foulbrood (AFB). The epidemiology of AFB is driven by the extreme spore resilience, the difficulty of bees to remove these spores, and the considerable incidence of undetected spore-producing colonies. The honeybee collective defence mechanisms and their feedback on colony development, which involves a division of labour at multiple levels of colony organization, are difficult to model. To better predict disease outbreaks we need to understand the feedback between colony development and disease progression within the colony. We therefore developed Bayesian models with data from forty AFB-diseased colonies monitored over an entire foraging season to (i) investigate the relationship between spore production and symptoms, (ii) disentangle the feedback loops between AFB epidemiology and natural colony development, and (iii) discuss whether larger insect societies promote or limit within-colony disease transmission.

Results

Rather than identifying a fixed spore count threshold for clinical symptoms, we estimated the probabilities around the relationship between spore counts and symptoms, taking into account modulators such as brood amount/number of bees and time post infection. We identified a decrease over time in the bees-to-brood ratio related to disease development, which should ultimately induce colony collapse. Lastly, two contrasting theories predict that larger colonies could promote either higher (classical epidemiological SIR-model) or lower (increasing spatial nest segregation and more effective pathogen removal) disease prevalence.

Conclusions

AFB followed the predictions of the SIR-model, partly because disease prevalence and brood removal are decoupled, with worker bees acting more as disease vectors, infecting new brood, than as agents of social immunity, by removing infected brood. We therefore established a direct link between disease prevalence and social group size for a eusocial insect. We furthermore provide a probabilistic description of the relationship between AFB spore counts and symptoms, and how disease development and colony strength over a season modulate this relationship. These results help to better understand disease development within honeybee colonies, provide important estimates for further epidemiological modelling, and gained important insights into the optimal sampling strategy for practical beekeeping and honeybee research.
Spontaneous fertility and variable spectrum of reproductive phenotype in a family with adult-onset X-linked adrenal insufficiency harboring a novel DAX-1/NR0B1 mutation
Authors: Vargas Michelle Cerutti C. , Moura Felipe Scipião , Elias Cecília P. , Carvalho Sara R. , Rassi Nelson , Kunii Ilda S. , Dias-da-Silva Magnus R. , Costa-Barbosa Flavia Amanda
Published: March 2020

Background

Adrenal hypoplasia congenita (AHC) is an X-linked disorder that affects the adrenal cortex and hypothalamus-pituitary-gonadal axis (HPG), leading to primary adrenocortical insufficiency (PAI) and hypogonadotropic hypogonadism. AHC is caused by a mutation in the DAX-1 gene (NR0B1). More commonly, this disease is characterized by early-onset PAI, with symptoms in the first months of life. However, a less severe phenotype termed late-onset AHC has been described, as PAI signs and symptoms may begin in adolescence and adulthood. Here we describe a family report of a novel mutation within NR0B1 gene and variable reproductive phenotypes, including spontaneous fertility, in a very late-onset X-linked AHC kindred.

Case presentation

Three affected maternal male relatives had confirmed PAI diagnosis between 30 y and at late 64 y. The X-linked pattern has made the endocrinology team to AHC suspicion. Regarding the HPG axis, all males presented a distinct degree of testosterone deficiency and fertility phenotypes, varying from a variable degree of hypogonadism, oligoasthenoteratozoospermia to spontaneous fertility. Interestingly, the other five maternal male relatives unexpectedly died during early adulthood, most likely due to undiagnosed PAI/adrenal crisis as the probable cause of their premature deaths. Sequencing analysis of the NR0B1 gene has shown a novel NR0B1 mutation (p.Tyr378Cys) that segregated in three AHC family members.

Conclusions

NR0B1 p.Tyr378Cys segregates in an AHC family with a variable degree of adrenal and gonadal phenotypes, and its hemizygous trait explains the disease in affected family members. We recommend that NR0B1 mutation carriers, even those that are allegedly asymptomatic, be carefully monitored while reinforcing education to prevent PAI and consider early sperm banking when spermatogenesis still viable.
In vitro anti-HIV activity of some Indian medicinal plant extracts
Authors: Palshetkar Aparna , Pathare Navin , Jadhav Nutan , Pawar Megha , Wadhwani Ashish , Kulkarni Smita , Singh Kamalinder K.
Published: March 2020

Background

Human Immunodeficiency Virus (HIV) persists to be a significant public health issue worldwide. The current strategy for the treatment of HIV infection, Highly Active Antiretroviral Therapy (HAART), has reduced deaths from AIDS related disease, but it can be an expensive regime for the underdeveloped and developing countries where the supply of drugs is scarce and often not well tolerated, especially in persons undergoing long term treatment. The present therapy also has limitations of development of multidrug resistance, thus there is a need for the discovery of novel anti-HIV compounds from plants as a potential alternative in combating HIV disease.

Methods

Ten Indian medicinal plants were tested for entry and replication inhibition against laboratory adapted strains HIV-1IIIB, HIV-1Ada5 and primary isolates HIV-1UG070, HIV-1VB59 in TZM-bl cell lines and primary isolates HIV-1UG070, HIV-1VB59 in PM1 cell lines. The plant extracts were further evaluated for toxicity in HEC-1A epithelial cell lines by transwell epithelial model.

Results

The methanolic extracts of Achyranthes aspera, Rosa centifolia and aqueous extract of Ficus benghalensis inhibited laboratory adapted HIV-1 strains (IC80 3.6–118 μg/ml) and primary isolates (IC80 4.8–156 μg/ml) in TZM-bl cells. Methanolic extract of Strychnos potatorum, aqueous extract of Ficus infectoria and hydroalcoholic extract of Annona squamosa inhibited laboratory adapted HIV-1 strains (IC80 4.24–125 μg/ml) and primary isolates (IC80 18–156 μg/ml) in TZM-bl cells. Methanolic extracts of Achyranthes aspera and Rosa centifolia, (IC801-9 μg/ml) further significantly inhibited HIV-1 primary isolates in PM1cells. Methanolic extracts of Tridax procumbens, Mallotus philippinensis, Annona reticulate, aqueous extract of Ficus benghalensis and hydroalcoholic extract of Albizzia lebbeck did not exhibit anti-HIV activity in all the tested strains. Methanolic extract of Rosa centifolia also demonstrated to be non-toxic to HEC-1A epithelial cells and maintained epithelial integrity (at 500 μg/ml) when tested in transwell dual-chamber.

Conclusion

These active methanolic extracts of Achyranthes aspera and Rosa centifolia, could be further subjected to chemical analysis to investigate the active moiety responsible for the anti-HIV activity. Methanolic extract of Rosa centifolia was found to be well tolerated maintaining the epithelial integrity of HEC-1A cells in vitro and thus has potential for investigating it further as candidate microbicide.
Neferine and lianzixin extracts have protective effects on undifferentiated caffeine-damaged PC12 cells
Authors: Chen Jingjing , Tang Mimi , Liu Manhua , Jiang Yueping , Liu Bin , Liu Shao
Published: March 2020

Background

The embryos of Nelumbo nucifera Gaertn seeds, lianzixin, are used in China as food and traditional herbal medicine. Principal therapeutic indications are insomnia, anxiety and pyrexia. Caffeine is a psychostimulant and excessive use predisposes to cell damage and neurotoxicity. We aimed to investigate the potential protect effect of Neferine and lianzixin extracts on undifferentiated caffeine-damaged phaeochromocytoma cells (PC12 cells).

Methods

A cell damage model based on undifferentiated PC12 was established with caffeine. Effect of Lianzixin extracts (total alkaloids, alcohol extract and water extract) and neferine on caffeine-damaged PC12 cells was evaluated. Cell viability was assessed using the methyl thiazolyl tetrazolium (MTT) assay, cellular morphology by inverted microscope, the nucleus by Hoechst 33342 staining and cleaved poly ADP-ribose polymerase (PARP) expression by western blot analysis.

Results

Lianzixin extracts (total alkaloids, alcohol extract and water extract) and neferine improved the viability of PC12 cells damaged by caffeine. The morphology of PC12 cells pretreated with neferine, or alcohol or water extract of lianzixin aggregated and attached better than caffeine-damaged cells, but cells pretreated with total alkaloids of lianzixin showed abnormal morphology. Compared with caffeine-damaged cells, cells pretreated with neferine, or alcohol or water extract of lianzixin showed a notable increase in nucleus staining and an obvious decrease in cleaved PARP expression.

Conclusions

Lianzixin extracts and neferine have protective effects against caffeine-induced damage in PC12 cells, which laid a foundation for finding a new medicine value of Lianzixin.
Introducing career skills for dental students as an undergraduate course at the University of Szeged, Hungary
Authors: Szabó Réka Magdolna , Davis Joan Mary , Antal Márk
Published: March 2020

Background

In the last three decades there is a growing recognition in the dental profession that dental education must go beyond teaching the technicalities of dentistry and include professionalism and communication skills that the future dentist may need. Such skills are best taught in a student-centered way. Literature suggests that student-centered elements are difficult to introduce in traditional, teacher-centered curricula. This is especially true in post-communist countries where higher education was under strict state control for decades. The aim of the piece of research presented here was to investigate how difficult it is to introduce a student-centered career skills course in a traditionally teacher-centered dental curriculum.

Methods

Considering the needs of our final-year dental students and Super’s model of career development, we created an undergraduate curricular career skills course running for two semesters in two languages (Hungarian and English). The primary aim of the course is to help students with their career expectations and develop their identity as a professional workforce. The secondary aim is to teach skills that students can use when applying for a job. At the end of the semesters, we assessed our students’ satisfaction with various aspects of the course by using a questionnaire. Results were analyzed item-wise and according to the main aspects of the course (i.e., groups of items organized around a particular aspect).

Results

General satisfaction with the course was high, and practical skills training (such as CV and motivation letter writing) got the highest scores. From the answers it appears that the students were the least comfortable with having to deal with their personal values and preferences.

Conclusions

While it is common for universities to offer various forms of career intervention, to our knowledge, no other university offers a curricular career skills course specifically for dental students. Our student-centered course designed in a problem-based learning framework worked even in a traditionally teacher-centered educational environment, where university students are rarely encouraged to be active participants in courses. By sharing our experience, we would like to encourage our fellow dental educators working in similar environments to devise and offer such courses.
Confidence and motivation to help those with a mental health problem: experiences from a study of nursing students completing mental health first aid (MHFA) training
Authors: Crawford Gemma , Burns Sharyn
Published: March 2020

Background

Those studying nursing are at greater risk for developing mental health problems than other tertiary students. Mental Health First Aid (MHFA) training may assist students to support peers and build mental health literacy. Understanding motivation to participate in training can identify factors influencing uptake and completion. This paper explores motivators for university nursing students to participate in MHFA training and uses previous experience and confidence in assisting someone with a mental health problem to triangulate data.

Method

A randomised controlled trial was employed to measure the impact of the course for nursing students at a large Western Australian university. An online survey was administered prior to MHFA training with undergraduate nursing students (n = 140). Thematic analysis of open-ended questions explores motivators to participate and help provided to an individual. Baseline frequencies describe demographics, confidence in helping and exposure to someone with a mental health problem. A Chi Square test compared confidence in helping and exposure to someone with a mental health problem.

Results

More than half of participants reported contact with individuals experiencing mental health problems (55%;n = 77); approximately a third (35.8%) reported limited confidence to assist. Those in previous contact with someone with a mental health problem (71.5%;n = 55) were significantly more likely to feel confident in helping (p = 0.044). Mental health literacy, helping others, career and experiences were described as training motivators.

Conclusion

Exploiting motivators, both intrinsic and extrinsic may increase MHFA training uptake and completion. Tertiary institutions would benefit from policy to embed MHFA training into nursing degrees. The training may have utility for university degrees more broadly.

Trial registration

Australian New Zealand Clinical Trials Registry;ACTRN12614000861651. Registered 11 August 2014 (retrospectively registered).
The relationship of nursing practice laws to suicide and homicide rates: a longitudinal analysis of US states from 2012 to 2016
Authors: Choi Kristen R. , Takada Sae , Saadi Altaf , Easterlin Molly C. , Buchbinder Liza S. , Natsui Shaw , Zimmerman Frederick J.
Published: March 2020

Background

Nursing resources can have a protective effect on patient outcomes, but nurses and nursing scope of practice have not been studied in relation to injury outcomes. The purpose of this study was to examine whether scope of practice and ease of practice laws for nurse practitioners and registered nurses are associated with suicide and homicide rates in the United States.

Methods

This state-level analysis used data from 2012 to 2016. The outcome variables were age-adjusted suicide and homicide rates. The predictor variables were NP scope of practice by state (limited, partial, or full) and RN ease of practice (state RN licensure compact membership status). Covariates were state sociodemographic, healthcare, and firearm/firearm policy context variables that have a known relationship with the outcomes.

Results

Full scope of practice for NPs was associated with lower rates of suicide and homicide, with stronger associations for suicide. Likewise, greater ease of practice for RNs was associated with lower suicide and homicide rates.

Conclusions

Findings suggest that nurses are an important component of the healthcare ecosystem as it relates to injury outcomes. Laws supporting full nursing practice may have a protective effect on population health in the area of injuries and future studies should explore this relationship further.
Efficiency of health resource utilisation in primary-level maternal and child health hospitals in Shanxi Province, China: a bootstrapping data envelopment analysis and truncated regression approach
Authors: Zhang Tao , Lu Wei , Tao Hongbing
Published: March 2020

Background

District- and county-level maternal and child health hospitals (MCHHs) are positioned to provide primary maternal and child healthcare in rural and urban areas of China. Their efficiencies and productivity largely affect the equity and accessibility of maternal and child health care. This study aimed to assess the efficiency of district- and county-level MCHHs in China and identify their associated factors.

Methods

Thirty-three district- and 84 county-level MCHHs were selected from Shanxi Province in 2017. At the first stage, bootstrapping data envelopment analysis (DEA) models were established to calculate the technical efficiency (TE), pure technical efficiency (PTE) and scale efficiency (SE) of district- and county-level hospitals. At the second stage, the estimated efficiency scores were regressed against external and internal hospital environmental factors by using bootstrap truncated regression to identify their determinants.

Results

The average TE, PTE and SE scores for district-level MCHHs were 0.7433, 0.8633 and 0.9335, respectively. All hospitals were found to be weakly efficient, although more than 50% of the hospitals performed with efficient SE (SE scores≥100%). As for county-level MCHHs, their average TE, PTE and SE scores were 0.5483, 0.6081 and 0.9329, respectively. The hospitals with TE and PTE scores less than 0.7 accounted for more than 60%, and no hospital was observed to operate effectively. Truncated regressions suggested that the proportion of health professionals, including doctors, nurses, pharmacists, inspection technician and image technician (district level:β = 0.57, 95% CI = 0.30–0.85; county level:β = 0.33, 95% CI = 0.15–0.52), and the number of health workers who received job training (district level:β = 0.67, 95% CI = 0.26–1.08; county level:β = 0.34, 95% CI = 0.14–0.54) had a positive association with efficiency scores. The amount of financial subsidy (β = 0.07, 95% CI = 0.05–0.09) was found to be directly proportional to the productive efficiency of the county-level MCHHs.

Conclusion

The operational inefficiency of district- and county-level MCHHs in Shanxi Province is severe and needs to be substantially improved, especially in terms of TE and PTE. Hiring additional medical personnel and ensuring the stability of the workforce should be prioritised. The Chinese government must provide sufficient financial subsidy to compensate for service costs.
Testing patient-informed approaches for visually depicting the hemoglobin A1c value to patients with poorly controlled diabetes: a randomized, controlled trial
Authors: Gopalan Anjali , Suttner Leah , Troxel Andrea B. , McDonough Kevin , Schapira Marilyn M.
Published: March 2020

Background

Patients’ understanding of the hemoglobin A1c (HbA1c) has been linked to better diabetes care outcomes (glycemic control, self-care). This is concerning given low documented rates of HbA1c understanding. In this non-blinded, randomized trial, we compared two formats for communicating the HbA1c, selected based on input from people with diabetes, to standard presentation to assess their impact on participants’ glycemic control and diabetes-related perceptions.

Methods

To design the tested formats, we interviewed 25 patients with diabetes and reviewed a range of possible formats, including color-based scales and graphs. The interviews were recorded, transcribed, and subjected to thematic analysis. Synthesizing interviewees’ feedback, we selected two formats, one using a combination of words and colors (Words) and one using a color-coded graph (Graph), for further evaluation. We then randomized adults with poorly controlled diabetes to receive mailed information on their current diabetes control in one of three ways: 1) standard lab report (control), 2) Words format, or 3) Graph format. The primary outcome was HbA1c change at 6 months. Also examined were changes in participants’ diabetes-related perceptions and choice of participation incentive.

Results

Of the 234 enrolled participants, 76.9% were Black, and their median baseline HbA1c was 9.1% (interquartile range 8.4–10.4). There were no between-arm differences in HbA1c change (− 1.04% [SD 2.2] Control vs. -0.59% [SD 2.0] Words vs. -0.54% [SD 2.1] Graph, p > 0.05 for all comparisons). Participants in the Words arm had an increase in the accuracy of their perceptions of diabetes seriousness (p = 0.04) and in the number of participants reporting a diabetes management goal (p = 0.01).

Conclusions

The two patient-informed communication formats did not differentially impact glycemic control among adults with inadequately controlled diabetes. However, a significant proportion of participants in the Words arm had an increase in the accuracy of their perception of diabetes seriousness, a potential mediating factor in positive diabetes-related behavioral changes. With increasing use of patient-facing online portals, thoughtfully designed approaches for visually communicating essential, but poorly understood, information like the HbA1c to patients have the potential to facilitate interpretation and support self-management.

Clinical trial registration

Prospectively registered as NCT01886170.
Complexity of nursing care at 24 h from admission predicts in-hospital mortality in medical units: a cohort study
Authors: Ausili Davide , Bernasconi Davide Paolo , Rebora Paola , Prestini Lucia , Beretta Giorgio , Ferraioli Laura , Cazzaniga Anna , Valsecchi Maria Grazia , Di Mauro Stefania
Published: March 2020

Background

The Informative System of Nursing Performance was developed to measure complexity of nursing care based on the actual interventions performed by nurses at the point of care. The association of this score with in-hospital mortality was not investigated before. Having this information is relevant to define evidence-based criteria that hospital administrators can use to allocate nursing workforce according to the real and current patients’ need for nursing care. The aim of this study is to assess the association between complexity of nursing care and in-hospital mortality.

Methods

Register-based cohort study on all patients admitted to acute medical wards of a middle-large hospital in the North of Italy between January 1, 2014, to December 31, 2015 and followed up to discharge. Out of all the eligible 7247 records identified in the Hospital Discharge Register, 6872 records from 5129 patients have been included. A multivariable frailty Cox model was adopted to estimate the association between the Informative System of Nursing Performance score, both as continuous variable and dichotomized as low (score < 50) or high (score ≥ 50), and in-hospital mortality adjusting for several factors recorded at admission (age, gender, type of admission unit, type of access and Charlson Comorbidity Index).

Results

The median age of the 5129 included patients was 76 [first-third quartiles 64–84] and 2657(52%) patients were males. Over the 6872 admissions, there were 395 in-hospital deaths among 2922 patients at high complexity of nursing care (13.5%) and 74/3950 (1.9%) among those at low complexity leading to a difference of 11.6% (95% CI: 10.3–13.0%). Adjusting by relevant confounders, the hazard rate of mortality in the first 10 days from admission resulted 6 times significantly higher in patients at high complexity of nursing care with respect to patients at low complexity (hazard ratio, HR 6.58, 95%CI: 4.50;9.62, p < 0.001). The HR was lower after 10 days from admission but still significantly higher than 1. By considering the continuous score, the association was confirmed.

Conclusion

Complexity of nursing care is strongly associated to in-hospital mortality of acute patients admitted to medical departments. It predicts in-hospital mortality better than widely used indicators, such as comorbidity.
Assessing health system challenges and opportunities for better noncommunicable disease outcomes: the case of Mauritius
Authors: Musango Laurent , Timol Maryam , Burhoo Premduth , Shaikh Faisal , Donnen Philippe , Kirigia Joses Muthuri
Published: March 2020

Background

The objectives of the study reported in this paper were: (a) to score the coverage of core NCD population-based interventions and individual services in Mauritius; (b) to analyse and score the presence of 15 common health system challenges that impede delivery of core NCD interventions and services in Mauritius; and (c) to provide policy recommendations for Mauritius to address health system barriers to delivery of NCD interventions and services.

Methods

The Mauritius country assessment applied the guidelines developed by the World Health Organization Regional Office for Europe for systematically scoring coverage of NCD interventions and assessing health system challenges for improving NCD outcomes. The assessment used qualitative research design approach.

Results

Of the 24 core population-based interventions for addressing key NCD risk factors, 16.7% were rated extensive, 37.5% moderate and 45.8% limited. Three (20%), 8 (53%) and 4 (27%) of the 15 individual/personal CVD, diabetes and cancer services were rated extensive, moderate and limited respectively. The top five health system challenges hampering scale-up of coverage of population-based NCD interventions in Mauritius were inadequate interagency cooperation; limited application of explicit priority setting approaches; inadequate change management; sub-optimal distribution and mix human resources; insufficient population empowerment; and insufficient political commitment. The top five challenges had average scores of between 3.1 (interagency cooperation) and 2.4 (distribution and mix of human resources).

The top five health system challenges constraining expansion in coverage of individual NCD services were limited integration of evidence into practice; limited use of explicit priority-setting approaches; inadequate application of information and technology solutions; insufficient population empowerment; and sub-optimal distribution and mix of human resources. The top five challenges for individual interventions had mean scores varying between 2.6 (integration of evidence into practice) and 1.7 (distribution and mix of human resources).

Conclusions

Mauritius needs to increase its domestic general government investments into the national health system and requisite multi-sectoral action to address the priority health system challenges with a view of bridging the existing gaps in coverage of NCD population-based interventions and individual services.
Prevalence, characteristics, and costs of diagnosed homocystinuria, elevated homocysteine, and phenylketonuria in the United States: a retrospective claims-based comparison
Authors: Sellos-Moura Marcia , Glavin Frank , Lapidus David , Evans Kristin , Lew Carolyn R. , Irwin Debra E.
Published: March 2020

Background

Classical homocystinuria (HCU), an inborn error of homocysteine metabolism, has previously been estimated to affect approximately 1 in 100,000–200,000 people in the United States (US). HCU is poorly detected by newborn screening, resulting in underestimates of its prevalence. This study compared characteristics, healthcare use and costs, and projected prevalence between patients with diagnosed HCU, elevated total homocysteine (tHcy), and diagnosed phenylketonuria (PKU).

Methods

Patients in the MarketScan® Research Databases were identified with strictly-defined HCU (> 2 diagnoses, including 1 ICD-10), broadly-defined HCU (> 1 ICD-10), elevated tHcy (> 20 μmol/L) without an HCU diagnosis, or > 1 ICD-9/ICD-10 PKU diagnosis during 1/1/2010–12/31/2016 (first qualifying claim = index). Demographics and healthcare utilization and costs per patient per month (PPPM) were compared between all cohorts, frequencies of comorbidities and medications were compared between HCU and elevated tHcy patients, and healthcare provider types were assessed among HCU patients. The prevalence of patients meeting each cohort definition was projected to the United States (US) population.

Results

Patients with strictly-defined (N = 2450) and broadly-defined (N = 6613) HCU, and with elevated tHcy (N = 2017), were significantly older than PKU patients (N = 5120) (57 vs. 56 vs. 53 vs. 18 years; p < 0.05). Vitamin D deficiency, hyperlipidemia, folic acid/B vitamins, and lipid-lowering medications, among others, were more common among diagnosed HCU patients vs. those with elevated tHcy (all p < 0.05). Rates of healthcare utilization were generally higher among HCU and elevated tHcy patients, compared to PKU, though total healthcare costs were similar between groups. Most HCU patients (~ 38%) received their index diagnosis from a primary care physician; very few (~ 1%) had any claim from a geneticist during their enrollment. The age-adjusted national prevalence of HCU was projected at 31,162 (95% CI: 30,411 – 31,913; ~ 1 in 10,000 of the US population) using the broad definition.

Conclusions

The actual prevalence of HCU may be > 10 times prior estimates, at 1 in 10,000 in the US, and this study suggests that HCU is not being diagnosed until later in life. Improvements to newborn screening, detection in young children, and physician education regarding HCU among patients may be necessary to alleviate the burden of this genetic disease.
Approaches to overcome flow cytometry limitations in the analysis of cells from veterinary relevant species
Authors: Hunka Julia , Riley John T. , Debes Gudrun F.
Published: March 2020

Background

Flow cytometry is a powerful tool for the multiparameter analysis of leukocyte subsets on the single cell level. Recent advances have greatly increased the number of fluorochrome-labeled antibodies in flow cytometry. In particular, an increase in available fluorochromes with distinct excitation and emission spectra combined with novel multicolor flow cytometers with several lasers have enhanced the generation of multidimensional expression data for leukocytes and other cell types. However, these advances have mainly benefited the analysis of human or mouse cell samples given the lack of reagents for most animal species. The flow cytometric analysis of important veterinary, agricultural, wildlife, and other animal species is still hampered by several technical limitations, even though animal species other than the mouse can serve as more accurate models of specific human physiology and diseases.

Results

Here we present time-tested approaches that our laboratory regularly uses in the multiparameter flow cytometric analysis of ovine leukocytes. The discussed approaches will be applicable to the analysis of cells from most animal species and include direct modification of antibodies by covalent conjugation or Fc-directed labeling (Zenon™ technology), labeled secondary antibodies and other second step reagents, labeled receptor ligands, and antibodies with species cross-reactivity.

Conclusions

Using refined technical approaches, the number of parameters analyzed by flow cytometry per cell sample can be greatly increased, enabling multidimensional analysis of rare samples and giving critical insight into veterinary and other less commonly analyzed species. By maximizing information from each cell sample, multicolor flow cytometry can reduce the required number of animals used in a study.
A de novo 2.2 Mb recurrent 17q23.1q23.2 deletion unmasks novel putative regulatory non-coding SNVs associated with lethal lung hypoplasia and pulmonary hypertension: a case report
Authors: Karolak Justyna A. , Gambin Tomasz , Honey Engela M. , Slavik Tomas , Popek Edwina , Stankiewicz Paweł
Published: March 2020

Background

Application of whole genome sequencing (WGS) enables identification of non-coding variants that play a phenotype-modifying role and are undetectable by exome sequencing. Recently, non-coding regulatory single nucleotide variants (SNVs) have been reported in patients with lethal lung developmental disorders (LLDDs) or congenital scoliosis with recurrent copy-number variant (CNV) deletions at 17q23.1q23.2 or 16p11.2, respectively.

Case presentation

Here, we report a deceased newborn with pulmonary hypertension and pulmonary interstitial emphysema with features suggestive of pulmonary hypoplasia, resulting in respiratory failure and neonatal death soon after birth. Using the array comparative genomic hybridization and WGS,two heterozygous recurrent CNV deletions: ~ 2.2 Mb on 17q23.1q23.2, involvingTBX4, and ~ 600 kb on 16p11.2, involvingTBX6,that both arose de novo on maternal chromosomes were identified. In the predicted lung-specific enhancer upstream toTBX4, we have detected seven novel putative regulatory non-coding SNVs that were absent in 13 control individuals with the overlapping deletions but without any structural lung anomalies.

Conclusions

Our findings further support a recently reported model of complex compound inheritance of LLDD in which both non-coding and coding heterozygousTBX4variants contribute to the lung phenotype. In addition, this is the first report of a patient with combined de novo heterozygous recurrent 17q23.1q23.2 and 16p11.2 CNV deletions.
Effect of average volume-assured pressure support treatment on health-related quality of life in COPD patients with chronic hypercapnic respiratory failure: a randomized trial
Authors: Magdy Doaa M. , Metwally Ahmed
Published: March 2020

The long-term effect of average volume-assured pressure support (AVAPS) on health-related quality of life (HRQOL) in chronic obstructive pulmonary disease (COPD) patients with chronic hypercapnic respiratory failure (CHRF) remains unclear.

The objective of this study is to identify the long-term effect of AVAPS in COPD patients with CHRF through assessment of HRQOL, exercise tolerance after six months duration.

Methods

In this randomized, controlled, parallel-group study, 40 stable hypercapnic COPD patients were randomized in a 1:1 ratio to receive either spontaneous timed AVAPS (ST/AVAPS) (intervention) or Bilevel positive airway pressure (ST/BiPAP) (control). HRQL was measured with the Short Form 12 Health Survey Questionnaire (SF-12). Exercise tolerance assessed by 6 min walking distance. Analyses were done between groups from baseline to the average of six months measurements.

Results

AVAPS led to significant 6 months improvements in several domains of (SF-12) compared to the control group, with the greatest improvement seen in general health [treatment effect of 8.2 points (95% confidence interval [95% CI 3.2 to 11.7; p = 0.001)], vitality (treatment effect 5.4 points [95% CI 1.4 to 9.3]; p = 0.001), physical functioning 5.5 points [95% CI 1.1 to 9.8]; p = 0.001) and bodily pain 5.1 points [95% CI 3.4 to 8.8]; p = 0.002). The physical health summary score improved by 3.7 points (95% CI 1.2 to 5.8; p = 0.001), but no significant improvement in the emotional or social role functioning, mental health subscale was noted.

AVAPS also resulted in improvement 6 min walking distance 9.2 points (95% CI − 1 to − 15];p = 0.001). A significant reduction in the daytime (PaCO2) was observed after 6 months in those treated with AVAPS.

Conclusions

In COPD patients with hypercapnic respiratory failure, AVAPS improved exercise tolerance and multiple domains of HRQOL over six months of follow-up, with the significant improvement observed in general health.
Production of ent-kaurene from lignocellulosic hydrolysate in Rhodosporidium toruloides
Authors: Geiselman Gina M. , Zhuang Xun , Kirby James , Tran-Gyamfi Mary B. , Prahl Jan-Philip , Sundstrom Eric R. , Gao Yuqian , Munoz Munoz Nathalie , Nicora Carrie D. , Clay Derek M. , Papa Gabriella , Burnum-Johnson Kristin E. , Magnuson Jon K. , Tanjore Deepti , Skerker Jeffrey M. , Gladden John M.
Published: March 2020

Background

Rhodosporidium toruloides has emerged as a promising host for the production of bioproducts from lignocellulose, in part due to its ability to grow on lignocellulosic feedstocks, tolerate growth inhibitors, and co-utilize sugars and lignin-derived monomers. Ent-kaurene derivatives have a diverse range of potential applications from therapeutics to novel resin-based materials.

Results

The Design, Build, Test, and Learn (DBTL) approach was employed to engineer production of the non-native diterpene ent-kaurene in R. toruloides. Following expression of kaurene synthase (KS) in R. toruloides in the first DBTL cycle, a key limitation appeared to be the availability of the diterpene precursor, geranylgeranyl diphosphate (GGPP). Further DBTL cycles were carried out to select an optimal GGPP synthase and to balance its expression with KS, requiring two of the strongest promoters in R. toruloides, ANT (adenine nucleotide translocase) and TEF1 (translational elongation factor 1) to drive expression of the KS from Gibberella fujikuroi and a mutant version of an FPP synthase from Gallus gallus that produces GGPP. Scale-up of cultivation in a 2 L bioreactor using a corn stover hydrolysate resulted in an ent-kaurene titer of 1.4 g/L.

Conclusion

This study builds upon previous work demonstrating the potential of R. toruloides as a robust and versatile host for the production of both mono- and sesquiterpenes, and is the first demonstration of the production of a non-native diterpene in this organism.
The multifaceted roles of long noncoding RNAs in pancreatic cancer: an update on what we know
Authors: Zhou Wenjia , Chen Lu , Li Chao , Huang Rui , Guo Mian , Ning Shangwei , Ji Jingjing , Guo Xiaorong , Lou Ge , Jia Xinqi , Zhao Junjie , Luo Feng , Li Chunlong , Qu Zhaowei , Yu Shan , Tai Sheng
Published: March 2020

Pancreatic cancer (PC) is one of the leading causes of cancer-related deaths worldwide. Due to the shortage of effective biomarkers for predicting survival and diagnosing PC, the underlying mechanism is still intensively investigated but poorly understood. Long noncoding RNAs (lncRNAs) provide biological functional diversity and complexity in protein regulatory networks. Scientific studies have revealed the emerging functions and regulatory roles of lncRNAs in PC behaviors. It is worth noting that some in-depth studies have revealed that lncRNAs are significantly associated with the initiation and progression of PC. As lncRNAs have good properties for both diagnostic and prognostic prediction due to their translation potential, we herein address the current understanding of the multifaceted roles of lncRNAs as regulators in the molecular mechanism of PC. We also discuss the possibility of using lncRNAs as survival biomarkers and their contributions to the development of targeted therapies based on the literature. The present review, based on what we know about current research findings, may help us better understand the roles of lncRNAs in PC.
Skp2 modulates proliferation, senescence and tumorigenesis of glioma
Authors: Wu Juan , Su Hong-kai , Yu Zhi-hui , Xi Shao-yan , Guo Cheng-cheng , Hu Zhe-yu , Qu Yue , Cai Hai-ping , Zhao Yi-ying , Zhao Hua-fu , Chen Fu-rong , Huang Yu-fan , To Shing-shun Tony , Feng Bing-hong , Sai Ke , Chen Zhong-ping , Wang Jing
Published: March 2020

Background

Gliomas represent the largest class of primary central nervous system neoplasms, many subtypes of which exhibit poor prognoses. Surgery followed by radiotherapy and chemotherapy has been used as a standard strategy but yielded unsatisfactory improvements in patient survival outcomes. The S-phase kinase protein 2 (Skp2), a critical component of the E3-ligase SCF complex, has been documented in tumorigenesis in various cancer types but its role in glioma has yet to be fully clarified. In this study, we investigated the function of Skp2 in the proliferation, stem cell maintenance, and drug sensitivity to temozolomide (TMZ) of glioma.

Methods

To investigate the role of Skp2 in the prognosis of patients with glioma, we first analyzed data in databases TCGA and GTEx. To further clarify the effect of Skp2 on glioma cell proliferation, we suppressed its level in glioblastoma (GBM) cell lines through knockdown and small molecule inhibitors (lovastatin and SZL-P1-41). We then detected cell growth, colony formation, sphere formation, drug sensitivity, and in vivo tumor formation in xenograft mice model.

Results

Skp2 mRNA level was higher in both low-grade glioma and GBM than normal brain tissues. The knockdown of Skp2 increased cell sensitivity to TMZ, decreased cell proliferation and tumorigenesis. In addition, Skp2 level was found increased upon stem cells enriching, while the knockdown of Skp2 led to reduced sphere numbers. Downregulation of Skp2 also induced senescence. Repurposing of lovastatin and novel compound SZL-P1-41 suppressed Skp2 effectively, and enhanced glioma cell sensitivity to TMZ in vitro and in vivo.

Conclusion

Our data demonstrated that Skp2 modulated glioma cell proliferation in vitro and in vivo, stem cell maintenance, and cell sensitivity to TMZ, which indicated that Skp2 could be a potential target for long-term treatment.
Circ_0032821 acts as an oncogene in cell proliferation, metastasis and autophagy in human gastric cancer cells in vitro and in vivo through activating MEK1/ERK1/2 signaling pathway
Authors: Jiang Yuanyuan , Zhang Yan , Chu Feifei , Xu Lidong , Wu Huili
Published: March 2020

Background

Circular RNA (circRNA) is increasingly attracting attention in gastric cancer (GC). Hsa_circ_0032821 (circ_0032821) has been declared to be upregulated in human GC tissues. However, the biological role of circ_0032821 remains undisclosed in GC cells.

Methods

Expression of circ_0032821 was measured by real-time quantitative PCR. Cell proliferation, autophagy, Epithelial-mesenchymal transition (EMT), migration, and invasion were evaluated by Cell counting kit-8 assay, western blotting or transwell assays. Expression of proliferating cell nuclear antigen (PCNA), Matrix metalloproteinase 2 (MMP2), MMP9, Light chain 3 (LC3), p62, total and phosphorylated Extracellular signal-regulated kinase 1/2 (ERK1/2) and Mitogen-activated protein kinase’s kinase 1 (MEK1) was evaluated by western blotting. Xenograft tumor model was established to measure tumor growth in vivo.

Results

Circ_0032821 was significantly upregulated in human GC tumors and cells. Moreover, circ_0032821 might be a biomarker for the advanced Tumor node metastasis (TNM) stage, lymphoid node metastasis and poor prognosis in gastric cancer. Knockdown of circ_0032821 by transfection induced decrease of cell proliferation, EMT, migration and invasion, but increase of autophagy of AGS and HGC-27 cells in vitro, as well as induced tumor growth inhibition in vivo. Besides, overexpression of circ_0032821 by transfection functioned the opposite effects in human GC cells. Mechanically, the MEK1/ERK1/2 signaling pathway was activated when circ_0032821 upregulation, whereas inhibited when circ_0032821 silencing.

Conclusion

Circ_0032821 expression induced cell proliferation, EMT, migration, invasion, and autophagy inhibition in human GC cells in vitro and in vivo through activating MEK1/ERK1/2 signaling pathway, suggesting circ_0032821 as an oncogenic role in GC.
Clarifying workforce flexibility from a division of labor perspective: a mixed methods study of an emergency department team
Authors: Wise Sarah , Duffield Christine , Fry Margaret , Roche Michael
Published: March 2020

Background

The need for greater flexibility is often used to justify reforms that redistribute tasks through the workforce. However, “flexibility” is never defined or empirically examined. This study explores the nature of flexibility in a team of emergency doctors, nurse practitioners (NPs), and registered nurses (RNs), with the aim of clarifying the concept of workforce flexibility. Taking a holistic perspective on the team’s division of labor, it measures task distribution to establish the extent of multiskilling and role overlap, and explores the behaviors and organizational conditions that drive flexibly.

Methods

The explanatory sequential mixed methods study was set in the Fast Track area of a metropolitan emergency department (ED) in Sydney, Australia. In phase 1, an observational time study measured the tasks undertaken by each role (151 h), compared as a proportion of time (Kruskal Wallis, Mann-WhitneyU), and frequency (Pearson chi-square). The time study was augmented with qualitative field notes. In phase 2, 19 semi-structured interviews sought to explain the phase 1 observations and were analyzed thematically.

Results

The roles were occupationally specialized: “Assessment and Diagnosis” tasks consumed the largest proportion of doctors’ (51.1%) and NPs’ (38.1%) time, and “Organization of Care” tasks for RNs (27.6%). However, all three roles were also multiskilled, which created an overlap in the tasks they performed. The team used this role overlap to work flexibly in response to patients’ needs and adapt to changing demands. Flexibility was driven by the urgent and unpredictable workload in the ED and enabled by the stability provided by a core group of experienced doctors and nurses.

Conclusion

Not every healthcare team requires the type of flexibility found in this study since that was shaped by patient needs and the specific organizational conditions of the ED. The roles, tasks, and teamwork that a team requires to “be flexible” (i.e., responsive and adaptable) are highly context dependent. Workforce flexibility therefore cannot be defined as a particular type of reform or role; rather, it should be understood as the capacity of a team to respond and adapt to patients’ needs within its organizational context. The study’s findings suggest that solutions for a more flexible workforce may lay in the organization of healthcare work.
SNP rs12982687 affects binding capacity of lncRNA UCA1 with miR-873-5p: involvement in smoking-triggered colorectal cancer progression
Authors: Fu Yang , Zhang Yizheng , Cui Jinyuan , Yang Ge , Peng Sanfei , Mi Wunan , Yin Xiangya , Yu Yang , Jiang Jianwu , Liu Qi , Qin Yiyu , Xu Wen
Published: March 2020

Background

This investigation was arranged to elucidate whether single nucleotide polymorphisms (SNPs) of lncRNA UCA1 was implicated in elevating colorectal cancer (CRC) risk by interacting with environmental exposures.

Methods

LncRNASNP database was firstly adopted to predict SNPs that possibly affected binding of UCA1 with miRNAs and then the interactive effect of SNPs and environmental exposure on CRC risk was evaluated by recurring to type 2 gene-environment interactions (GEI) model. Besides, MTT assay, colony formation assay, transwell assay and wound healing assay were performed to assess the activity of CRC cell lines which carried distinct genotypes of specific SNPs. The impact of nicotine on activity of CRC cells was also appraised.

Results

SNP rs12982687 of UCA1 intervened in the binding capacity of UCA1 with several miRNAs, especially miR-873-5p. MiRNAs regulated by UCA1, as predicted by mirPath software, shared genes that were enriched in HIF1 signaling pathway. Moreover, homozygote TT of rs12982687 reduced CRC risk among smokers, and CRC cells that carried rs12982687 (CC) displayed strong migration and invasion. By contrast, miR-873-5p mimic, which reduced UCA1 expression, delayed metastasis of CRC cells (all P < 0.05). Additionally, nicotine not merely elevated UCA1 and HIF-1α expressions in CRC cells, but also facilitated proliferation and metastasis of CRC cells (P < 0.05).

Conclusions

SNP rs12982687 was involved in smoking-triggered CRC progression, given its influence on UCA1's binding with miR-873-5p and HIF-1 signaling.