bluuebunny/crossref_metadata_2025 huggingface dataset:
An oral combined contraceptive user with elevated D-dimer post COVID-19: a case report
Authors: Alyousefi Nada A.
Published: August 2021

Background

This case discusses the challenges created by COVID-19 (coronavirus disease 2019) in the area of hormonal contraception, highlighting the contraception knowledge gap for women in their post COVID-19 period, especially if they had high D-dimer levels.

Case presentation

This case involves a thirty-eight-year-old woman taking combined oral contraception (desogestrel/ethinyl oestradiol tablets) with a history of varicose veins. She recovered from a COVID-19 infection in November 2020. She presented to the emergency room with right lower-limb pain below the knee and progressive swelling for five days in February 2021. Physical examination of the lower limb showed mild swelling and tenderness of the right leg compared to the left leg. D-Dimer was elevated (1.06 mcg/mL FEU). COVID-19 screening was negative. A Doppler scan to exclude DVT was performed considering the clinical picture and high D-dimer level. There was no evidence of DVT in the right limb. She was reassured and discharged with instructions on when to visit the emergency room. The D-dimer had decreased to 0.53 mcg/mL FEU in March 2021. She booked an appointment with family medicine clinics because she was concerned about the continuation of combined oral contraception (desogestrel/ethinyl oestradiol tablets) with high D-dimer and risk of thrombosis. The follow-up D-dimer level in May 2021 was normal (0.4 mcg/mL FEU). The patient preferred to continue taking oral contraception.

Conclusion

An evidence-based consensus is needed to guide clinicians in providing contraception counselling for such patients.
Indole alkaloids inhibit zika and chikungunya virus infection in different cell lines
Authors: Monsalve-Escudero Laura Milena , Loaiza-Cano Vanessa , Pájaro-González Yina , Oliveros-Díaz Andrés Felipe , Diaz-Castillo Fredyc , Quiñones Wiston , Robledo Sara , Martinez-Gutierrez Marlen
Published: August 2021

Background

In recent years, an increase in the occurrence of illnesses caused by two clinically- important arboviruses has been reported: Zika virus (ZIKV) and Chikungunya virus (CHIKV). There is no licensed antiviral treatment for either of the two abovementioned viruses. Bearing in mind that the antiviral effect of indole alkaloids has been reported for other arboviral models, the present study proposed to evaluate the antiviral in vitro and in silico effects of four indole alkaloids on infections by these two viruses in different cell lines.

Methods

The antiviral effects of voacangine (VOAC), voacangine-7-hydroxyindolenine (VOAC-OH), rupicoline and 3-oxo voacangine (OXO-VOAC) were evaluated in Vero, U937 and A549 cells using different experimental strategies (Pre, Trans, Post and combined treatment). Viral infection was quantified by different methodologies, including infectious viral particles by plating, viral genome by RT-qPCR, and viral protein by cell ELISA. Moreover, molecular docking was used to evaluate the possible interactions between structural and nonstructural viral proteins and the compounds. The results obtained from the antiviral strategies for each experimental condition were compared in all cases with the untreated controls. Statistically significant differences were identified using a parametric Student’s t-test. In all cases, p values below 0.05 (p < 0.05) were considered statistically significant.

Results

In the pre-treatment strategy in Vero cells, VOAC and VOAC-OH inhibited both viral models and OXO-VOAC inhibited only ZIKV; in U937 cells infected with CHIKV/Col, only VOAC-OH inhibited infection, but none of the compounds had activity in A549 cells; in U937 cells and A549 cells infected with ZIKV/Col, the three compounds that were effective in Vero cells also had antiviral activity. In the trans-treatment strategy, only VOAC-OH was virucidal against ZIKV/Col. In the post-treatment strategy, only rupicoline was effective in the CHIKV/Col model in Vero and A549 cells, whereas VOAC and VOAC-OH inhibited ZIKV infection in all three cell lines. In the combined strategy, VOAC, VOAC-OH and rupicoline inhibited CHIKV/Col and ZIKV/Col, but only rupicoline improved the antiviral effect of ZIKV/Col-infected cultures with respect to the individual strategies. Molecular docking showed that all the compounds had favorable binding energies with the structural proteins E2 and NSP2 (CHIKV) and E and NS5 (ZIKV).

Conclusions

The present study demonstrates that indole alkaloids are promising antiviral drugs in the process of ZIKV and CHIKV infection; however, the mechanisms of action evaluated in this study would indicate that the effect is different in each viral model and, in turn, dependent on the cell line.
Stress and behavior patterns throughout medical education – a six year longitudinal study
Authors: Voltmer Edgar , Köslich-Strumann Susen , Voltmer Jan-Bennet , Kötter Thomas
Published: August 2021

Background

Medical education has a reputation for being demanding and stressful. However, longitudinal surveys across the whole course of study considering risks and resources are rare.

Methods

For the evaluation of stress and coping we administered the standard instruments Perceived Medical School Stress Scale (PMSS), Hospital Anxiety and Depression Scale (HADS), Work-Related Behavior and Experience Patterns (AVEM), Maslach Burnout Inventory (MBI), and a short form of the Coping Orientations to Problems Experienced Scale (Brief COPE) in three consecutive cohorts of medical students (N = 377) at one German university. Students were surveyed at the beginning of their studies (t0) and again during each consecutive summer semester (t1-t6).

Results

Stress and symptoms of anxiety and depression increased in the first two years of medical studies but decreased again towards their end. Consistently, freshmen medical students presented with a large proportion of the healthy pattern at t0 (56 %) that decreased to 30 % at t2, and increased up to 44 % at t6. Correspondingly, the proportion with the burnout-related risk pattern B increased from 9 to 16 % at t2, again decreasing to 7 % at t6. Over the whole course of study there was an almost continuous increase of the unambitious pattern S from t0 13 to 40 % at t6. Characteristic differences especially between the healthy pattern and the risk patterns regarding stress, mental health symptoms and coping were observed. Female students showed a higher vulnerability for stress, anxiety and depression as well as lower proportions with a healthy pattern, and higher proportions with risk patterns for overexertion and burnout.

Conclusions

The development of stress, symptoms and behavior and experience patterns especially in the first two years, demonstrating increasing study-related stress in the preclinical years, as well as the high proportion with an unambitious pattern at the end of the course of study emphasize the need for prevention and health promotion at both the individual and contextual levels.
What works for whom in compassion training programs offered to practicing healthcare providers: a realist review
Authors: Sinclair Shane , Kondejewski Jane , Jaggi Priya , Roze des Ordons Amanda L. , Kassam Aliya , Hayden K. Alix , Harris Daranne , Hack Thomas F.
Published: August 2021

Background

Patients and families want their healthcare to be delivered by healthcare providers that are both competent and compassionate. While compassion training has begun to emerge in healthcare education, there may be factors that facilitate or inhibit the uptake and implementation of training into practice. This review identified the attributes that explain the successes and/or failures of compassion training programs offered to practicing healthcare providers.

Methods

Realist review methodology for knowledge synthesis was used to consider the contexts, mechanisms (resources and reasoning), and outcomes of compassion training for practicing healthcare providers to determine what works, for whom, and in what contexts.

Results

Two thousand nine hundred ninety-one articles underwent title and abstract screening, 53 articles underwent full text review, and data that contributed to the development of a program theory were extracted from 45 articles. Contexts included the clinical setting, healthcare provider characteristics, current state of the healthcare system, and personal factors relevant to individual healthcare providers. Mechanisms included workplace-based programs and participatory interventions that impacted teaching, learning, and the healthcare organization. Contexts were associated with certain mechanisms to effect change in learners’ attitudes, knowledge, skills and behaviors and the clinical process.

Conclusions

In conclusion this realist review determined that compassion training may engender compassionate healthcare practice if it becomes a key component of the infrastructure and vision of healthcare organizations, engages institutional participation, improves leadership at all levels, adopts a multimodal approach, and uses valid measures to assess outcomes.
Medical education in china: progress in the past 70 years and a vision for the future
Authors: Wang Weimin
Published: August 2021

Medical education in China has undergone significant reforms in contemporary times. As the world’s largest medical education system, it is important to understand the status of China’s medical education in our interdependent world. This paper highlights the current landscape of medical education in China, particularly the progress that have been made in recent years. It also examines the current topics and challenges facing China’s medical educators today, and proposed recommendations for improving medical education in China. The medical education in China will produce better qualified health professionals to meet the health needs of Chinese population according to the new requirements of the “Healthy China 2030” blueprint.
Online versus classroom teaching for medical students during COVID-19: measuring effectiveness and satisfaction
Authors: AlQhtani Abdullh , AlSwedan Nasser , Almulhim Abdullelah , Aladwan Raghad , Alessa Yara , AlQhtani Kholoud , Albogami Malak , Altwairqi Khalid , Alotaibi Fahad , AlHadlaq Abdulmajeed , Aldhafian Osama
Published: August 2021

Background

The COVID-19 pandemic and physical distancing have had a significant impact on the conversion of traditional teaching methods to online teaching methods, which although not uncommon in medical schools, has to date only been used for some aspects of the teaching process. Thus, we aimed to measure the effectiveness of e-learning during the COVID-19 pandemic, as well as medical students’ preferences regarding e-learning and classroom teaching, and the possibility of applying it post-pandemic.

Methods

A cross-sectional online survey of medical students (N = 376) in six medical schools was carried out after their second semester, from August 15 to 20, 2020. Ten parameters were measured for the effectiveness of e-learning based on a 5-point Likert-scale and five parameters were measured for satisfaction.

Results

e-learning was more or equally effective in four parameters such as assignment submission and meeting individual needs, but less effective in six parameters, including building skills and knowledge, and interaction level. Satisfaction was either high or neutral in all five parameters.

Conclusions

Our findings have shown that e-learning can assist the teaching process in medical schools in some respects, but cannot be used for the entire teaching process.
Evaluating an integrated care pathway for frail elderly patients in Norway using multi-criteria decision analysis
Authors: Islam M. Kamrul , Ruths Sabine , Jansen Kristian , Falck Runa , Mölken Maureen Rutten-van , Askildsen Jan Erik
Published: August 2021

Background

To provide value-based care for patients with multi-morbidity, innovative integrated care programmes and comprehensive evaluations of such programmes are required. In Norway, a new programme called “Holistic Continuity of Patient Care” (HCPC) addresses the issue of multi-morbidity by providing integrated care within learning networks for frail elderly patients who receive municipal home care services or a short-term stay in a nursing home. This study conducts a multi-criteria decision analysis (MCDA) to evaluate whether the HCPC programme performs better on a large set of outcomes corresponding to the ‘triple aim’ compared to usual care.

Methods

Prospective longitudinal survey data were collected at baseline and follow-up after 6-months. The assessment of HCPC was implemented by a novel MCDA framework. The relative weights of importance of the outcomes used in the MCDA were obtained from a discrete choice experiment among five different groups of stakeholders. The performance score was estimated using a quasi-experimental design and linear mixed methods. Performance scores were standardized and multiplied by their weights of importance to obtain the overall MCDA value by stakeholder group.

Results

At baseline in the HCPC and usual care groups, respectively, 120 and 89 patients responded, of whom 87 and 41 responded at follow-up. The average age at baseline was 80.0 years for HCPC and 83.6 for usual care. Matching reduced the standardized differences between the groups for patient background characteristics and outcome variables. The MCDA results indicated that HCPC was preferred to usual care irrespective of stakeholders. The better performance of HCPC was mostly driven by improvements in enjoyment of life, psychological well-being, and social relationships and participation. Results were consistent with sensitivity analyses using Monte Carlo simulation.

Conclusion

Frail elderly with multi-morbidity represent complex health problems at large costs for society in terms of health- and social care. This study is a novel contribution to assessing and understanding HCPC programme performance respecting the multi-dimensionality of desired outcomes. Integrated care programmes like HCPC may improve well-being of patients, be cost-saving, and contribute to the pursuit of evidence based gradual reforms in the care of frail elderly.
Utilisation and safety of catheter ablation of atrial fibrillation in public and private sector hospitals
Authors: Ngo Linh , Ali Anna , Ganesan Anand , Woodman Richard J , Adams Robert , Ranasinghe Isuru
Published: August 2021

Background

Little is known about the utilisation and safety of catheter ablation of atrial fibrillation (AF) among public and private sector hospitals.

Aims

To examine the uptake of AF ablations and compare procedural safety between the sectors.

Method:

Hospitalisation data from all public and private hospitals in four large Australian states (NSW, QLD, VIC and WA) were used to identify patients undergoing AF ablation from 2012 to 17. The primary endpoint was any procedure-related complications up to 30-days post-discharge. Logistic regression was used to evaluate the association between treatment at a public hospital and risk of complications adjusting for covariates.

Results

Private hospitals performed most of the 21,654 AF ablations identified (n = 16,992, 78.5 %), on patients who were older (63.5 vs. 59.9y) but had lower rates of heart failure (7.9 % vs. 10.4 %), diabetes (10.2 % vs. 14.1 %), and chronic kidney diseases (2.4 % vs. 5.2 %) (all p < 0.001) than those treated in public hospitals. When compared with private hospitals, public hospitals had a higher crude rate of complications (7.25 % vs. 4.70 %, p < 0.001). This difference remained significant after adjustment (OR 1.74 [95 % CI 1.54–2.04]) and it occurred with both in-hospital (OR 1.83 [1.57–2.14]) and post-discharge (OR 1.39 [1.06–1.83]) complications, with certain complications including acute kidney injury (OR 5.31 [3.02–9.36]), cardiac surgery (OR 5.18 [2.19–12.27]), and pericardial effusion (OR 2.18 [1.50–3.16]).

Conclusions

Private hospitals performed most of AF ablations in Australia with a lower rate of complications when compared with public hospitals. Further investigations are needed to identify the precise mechanisms of this observed difference.
Association between service scope of primary care facilities and patient outcomes: a retrospective study in rural Guizhou, China
Authors: Li Zhong , Shi Meng , He Ruibo , Zhang Mei , Zhang Chi , Xiong Xinyu , Zhang Liang , Li Boyang
Published: August 2021

Background

Extending service scope of primary care facilities (PCFs) has been widely concerned in China. However, no current data about association between service scope of PCFs with patient outcomes are available. This study aims to investigate association between service scope of PCFs and patient outcomes.

Methods

A multistage, stratified clustered sampling method was used to collect information about service scope of PCFs from rural Guizhou, China. Claim data of 299,633 inpatient cases covered by 64 PCFs were derived from local information system of New Rural Cooperation Medical Scheme. Service scope of PCFs was collected with self-administrated questionnaires. Primary outcomes were (1) level of inpatient institutions, (2) length of stay, (3) per capita total health cost, (4) per capita out-of-pocket cost, (5) reimbursement ratio, (6) 30-day readmission. A total of 64 PCFs were categorized into five groups per facility-level service scope scores. Generalized linear regression models, logistic regression model, and ordinal regression model were conducted to identify association between service scope of PCFs and patient outcomes.

Results

On average, the median service scope score of PCFs was 20, with wide variation across PCFs. After controlling for demographic and clinical characteristics, patients living in communities with PCFs of greatest service scope (Quintile V vs. I) tended to have smaller rates of admission by county-level hospitals (-6.2 % [-6.5 %, -5.9 %], city-level hospitals (-1.9 % [-2.0 %, -1.8 %]), and provincial hospitals (-2.1 % [-2.2 %, -2.0 %]), smaller rate of 30-day readmission (-0.5 % [-0.7 %, -0.2 %]), less total health cost (-201.8 [-257.9, -145.8]) and out-of-pocket cost (-210.2 [-237.2, -183.2]), and greater reimbursement ratio (2.3 % [1.9 %, 2.8 %]) than their counterparts from communities with PCFs of least service scope.

Conclusions

Service scope of PCFs varied a lot in rural Guizhou, China. Greater service scope was associated with a reduction in secondary and tertiary hospital admission, reduced total cost and out-of-pocket cost, and 30-day readmission and increased reimbursement ratio. These results raised concerns about access to care for patients discharged from hospitals, which suggests potential opportunities for cost savings and improvement of quality of care. However, further evidence is warranted to investigate whether extending service scope of PCFs is cost-effective and sustainable.
Medical service utilization and out-of-pocket spending among near-poor National Health Insurance members in South Korea
Authors: Park Sooyeol
Published: August 2021

Background

The public health care system in South Korea is a two-tiered system. The lowest-income population is covered by the Medical Aid program, and the remaining population is covered by the National Health Insurance. The near poor, a relatively low-income population which is excluded from South Korea’s Medical Aid program due to exceeding the income threshold, experiences insufficient use of medical services and incurs high out-of-pocket expenses due to a lack of coverage under the country’s National Health Insurance (NHI) program. This study aims to examine medical utilization, out-of-pocket spending, and the occurrence of catastrophic health expenditures among the near-poor group compared to both Medical Aid beneficiaries and other (higher income) NHI members.

Methods

A cross-sectional study was conducted drawing upon a nationally representative dataset derived from the 2018 Korea Welfare Panel Study. The study classified people into three groups: Medical Aid beneficiaries; the near-poor population below 50 % of the median income threshold but still not qualifying for Medical Aid and thus enrolled in NHI; and NHI members above the threshold of 50 % of the median income. Using a generalized boosted model to estimate the propensity score weights between study groups, this study examined medical utilization, out-of-pocket spending, and the occurrence of catastrophic health expenditure among the study groups.

Results

The findings suggest that the utilization of medical services was not significantly different among the study groups. However, out-of-pocket spending and the occurrence of catastrophic health expenditure were significantly higher in the near-poor group compared to the other two groups.

Conclusions

The study found that the near-poor group was the most vulnerable among the Korean population because of their higher chance of incurring greater out-of-pocket spending and catastrophic health expenditures than is the case among the Medical Aid beneficiary and above-poverty line groups. Health policy needs to take the vulnerability of this near-poor population into account.
Lean adoption in hospitals: the role of contextual factors and introduction strategy
Authors: Rosa Angelo , Marolla Giuliano , Lega Federico , Manfredi Francesco
Published: August 2021

Background

In the scientific literature, many studies describe the application of lean methodology in the hospital setting. Most of the articles focus on the results rather than on the approach adopted to introduce the lean methodology. In the absence of a clear view of the context and the introduction strategy, the first steps of the implementation process can take on an empirical, trial and error profile. Such implementation is time-consuming and resource-intensive and affects the adoption of the model at the organizational level. This research aims to outline the role contextual factors and introduction strategy play in supporting the operators introducing lean methodology in a hospital setting.

Methodology

The methodology is revealed in a case study of an important hospital in Southern Italy, where lean has been successfully introduced through a pilot project in the pathway of cancer patients. The originality of the research is seen in the detailed description of the contextual elements and the introduction strategy.

Results

The results show significant process improvements and highlight the spontaneous dissemination of the culture of change in the organization and the streamlined adoption at the micro level.

Conclusion

The case study shows the importance of the lean introduction strategy and contextual factors for successful lean implementation. Furthermore, it shows how both factors influence each other, underlining the dynamism of the organizational system.
Patient characteristics and outcome in three different working models of home-based rehabilitation: a longitudinal observational study in primary health care in Norway
Authors: Meisingset Ingebrigt , Bjerke Joakim , Taraldsen Kristin , Gunnes Mari , Sand Sylvi , Hansen Anne E. , Myhre Gard , Evensen Kari Anne I.
Published: August 2021

Background

The organisation of health care services for older adults varies within and between countries. In Norway, primary care physiotherapy services offer home-based rehabilitation to older adults. The aim of this study was to compare patients’ characteristics and treatment outcomes in three working models of home-based rehabilitation.

Methods

Patients referred to home-based rehabilitation in a large municipality in Norway were invited to participate in this prospective observational study. The three working models, early intervention, reablement and regular physiotherapy, were organised according to the patients’ function and degree of independence. The older adults (≥ 65 years) were allocated to the different models by either a multidisciplinary group of health care personnel or by direct referral. Patients’ demographic and clinical characteristics, including physical function (Patient-specific functional scale, PSFS), physical performance (Short Physical Performance Battery, SPPB) and health-related quality of life (EQ-5D) were registered at baseline and follow-up (maximum 6 months after baseline). One-way ANOVA was used to analyse group differences in clinical characteristics and paired t-tests to analyse changes from baseline to follow-up.

Results

In total, 603 and 402 patients (median (interquartile range) age: 84 (77–88) years) completed baseline and follow-up assessments, respectively. Patients in all three working models had an increased risk for functional decline. Patients receiving early intervention (n = 62) had significantly (p < 0.001) better physical performance and health-related quality of life (SPPB mean 7.9, SD 2.7; EQ-5D:mean 0.59, SD 0.19), than patients receiving reablement (n = 132) (SPPB: mean 5.5, SD 2.6; EQ-5D: mean 0.50, SD 0.15) and regular physiotherapy (n = 409) (SPPB: mean 5.6, SD 2.8; EQ-5D: mean 0.41, SD 0.22). At follow-up, the three working models showed significantly improvements in physical function (PSFS: mean change (95 % CI): 2.5 (1.9 to 3.2); 1.8 (0.5 to 3.1); 1.7 (0.8 to 2.6), for regular physiotherapy, reablement, and early intervention, respectively). Patients receiving regular physiotherapy and reablement also significantly improved physical performance and health-related quality of life.

Conclusions

While older adults receiving reablement and regular physiotherapy showed similar patient characteristics and treatment outcomes, early intervention identified older patients at risk of functional decline at an earlier stage. These results are relevant for policy makers when designing and improving prevention and rehabilitation strategies in primary health care.
A community-based mobile clinic model delivering PrEP for HIV prevention to adolescent girls and young women in Cape Town, South Africa
Authors: Rousseau Elzette , Bekker Linda-Gail , Julies Robin F. , Celum Connie , Morton Jennifer , Johnson Rachel , Baeten Jared M. , O’Malley Gabrielle
Published: August 2021

Background

Daily doses of pre-exposure prophylaxis (PrEP) can reduce the risk of acquiring HIV by more than 95 %. In sub-Saharan Africa, adolescent girls and young women (AGYW) are at disproportionately high risk of acquiring HIV, accounting for 25 % of new infections. There are limited data available on implementation approaches to effectively reach and deliver PrEP to AGYW in high HIV burden communities.

Methods

We explored the feasibility and acceptability of providing PrEP to AGYW (aged 16–25 years) via a community-based mobile health clinic (CMHC) known as the Tutu Teen Truck (TTT) in Cape Town, South Africa. The TTT integrated PrEP delivery into its provision of comprehensive sexual and reproductive health services (SRHS). We analyzed data from community meetings and in-depth interviews with 30 AGYW PrEP users to understand the benefits and challenges of PrEP delivery in this context.

Results

A total of 585 young women started PrEP at the TTT between July 2017 – October 2019. During in-depth interviews a subset of 30 AGYW described the CMHC intervention for PrEP delivery as acceptable and accessible. The TTT provided services at times and in neighborhood locations where AGYW organically congregate, thus facilitating service access and generating peer demand for PrEP uptake. The community-based nature of the CMHC, in addition to its adolescent friendly health providers, fostered a trusting provider-community-client relationship and strengthened AGYW HIV prevention self-efficacy. The integration of PrEP and SRHS service delivery was highly valued by AGYW. While the TTT’s integration in the community facilitated acceptability of the PrEP delivery model, challenges faced by the broader community (community riots, violence and severe weather conditions) also at times interrupted PrEP delivery.

Conclusions

PrEP delivery from a CMHC is feasible and acceptable to young women in South Africa. However, to effectively scale-up PrEP it will be necessary to develop diverse PrEP delivery locations and modalities to meet AGYW HIV prevention needs.
Mineral status and enteric methane production in dairy cows during different stages of lactation
Authors: Grešáková Ľubomíra , Holodová Monika , Szumacher-Strabel Małgorzata , Huang Haihao , Ślósarz Piotr , Wojtczak Janusz , Sowińska Natalia , Cieślak Adam
Published: August 2021

Background

Lactating dairy cows are the greatest livestock contributor of methane, a major global greenhouse gas (GHG). However, good feeding management with adequate mineral intake can offers an effective approach to maintaining high levels of milk production and the health of dairy cows over the entire course of lactation, while also helping to reduce methane emission. The study described here investigated the plasma concentrations of both macroelements (Ca, Na, K, Mg, P) and microelements (Zn, Cu, Fe, Mn), as well as enteric methane emission and milk composition in high-yielding dairy cows in different lactation periods. The experiment was performed on Holstein–Friesian dairy cows with the average milk yield of 41 (± 9) L/day in a Polish commercial farm with modern dairy systems. A total of thirty high-yielding dairy cows were randomly assigned into three groups differing by lactation stage: early stage (Early, days 25–100), middle stage (Middle, days 101–250), and late stage (Late, day 250 and later). Dietary treatment for all cows was a total mixture ration (TMR) with maize and alfalfa silage the main forage components.

Results

The greatest milk yield and methane production were recorded in early-stage lactating cows, but the greatest methane intensity per kg of corrected milk was recorded in the late stage of lactation. Plasma concentrations of macroelements and microelements did not differ by lactation stages, but increased plasma concentrations of Zn and Fe and decreased plasma levels of Mg were noted during lactation. A positive correlation was found between plasma levels of Mg and other macroelements (Ca, Na, K), and between the concentrations of Fe and Zn, P in plasma, but no correlation between methane emission and mineral status was detected in the different lactation stages.

Conclusions

Our results showed different mineral requirements and enteric methane emissions in each lactation stage. The feeding strategy and mineral utilization were adequate to maintain the health, mineral status, and milk production of the Holstein cows during the entire lactation period, and suggest an effective way of reducing methane emission.
Recombinant expression of Barnase in Escherichia coli and its application in plasmid purification
Authors: Shankar Ram , Schäffer Nina , Schmeer Marco , Risse Joe Max , Friehs Karl , Schleef Martin
Published: August 2021

Background

The use of bovine-origin ribonucleases has been part of the standard protocol for plasmid DNA purification. As the field of gene therapy now enters the clinical stage, such enzymes need to be phased out or alternative purification protocols need to be developed to ensure product safety and regulatory compliance. The recombinant expression of bacterial RNase is fraught with toxicity problems making it a challenging enzyme to express. The current study describes a plasmid construct that allowed expression of barnase in Escherichia coli under co-expression of its native inhibitor barstar.

Results

The pure enzyme without the inhibitor barstar was exported to the extracellular space through the periplasm and then purified from the cell-free supernatant. Cation exchange chromatography was employed as a primary purification step. This was followed by hydrophobic interaction chromatography which resulted in a concentrated fraction of active enzyme. Although current levels of volumetric activity achieved are quite meagre (4 Kunitz units mL− 1), in principle its application to plasmid DNA purification could be proved. Currently, this is capable of processing small amounts (13 g) of bacterial biomass for plasmid production.

Conclusions

The current work focusses on the downstream purification strategies for a recombinant RNase and sets a framework for higher scale production if specific productivity is increased by optimal hosts and/or re-engineered plasmids. Also important is to curtail the massive enzyme loss during purification by cation exchange chromatography. Application of even a relatively small amount of recombinant RNase would contribute to greatly reducing the initial RNA levels in alkaline lysates thereby augmenting further downstream plasmid purification steps.
Combining protein and metabolic engineering strategies for biosynthesis of melatonin in Escherichia coli
Authors: Zhang Yanfeng , He Yongzhi , Zhang Nan , Gan JiaJia , Zhang Shan , Dong Zhiyang
Published: August 2021

Background

Melatonin has attracted substantial attention because of its excellent prospects for both medical applications and crop improvement. The microbial production of melatonin is a safer and more promising alternative to chemical synthesis approaches. Researchers have failed to produce high yields of melatonin in common heterologous hosts due to either the insolubility or low enzyme activity of proteins encoded by gene clusters related to melatonin biosynthesis.

Results

Here, a combinatorial gene pathway for melatonin production was successfully established in Escherichia coli by combining the physostigmine biosynthetic genes from Streptomyces albulus and gene encoding phenylalanine 4-hydroxylase (P4H) from Xanthomonas campestris and caffeic acid 3-O-methyltransferase (COMT) from Oryza sativa. A threefold improvement of melatonin production was achieved by balancing the expression of heterologous proteins and adding 3% glycerol. Further protein engineering and metabolic engineering were conducted to improve the conversion of N-acetylserotonin (NAS) to melatonin. Construction of COMT variant containing C303F and V321T mutations increased the production of melatonin by fivefold. Moreover, the deletion of speD gene increased the supply of S-adenosylmethionine (SAM), an indispensable cofactor of COMT, which doubled the yield of melatonin. In the final engineered strain EcMEL8, the production of NAS and melatonin reached 879.38 ± 71.42 mg/L and 136.17 ± 1.33 mg/L in a shake flask. Finally, in a 2-L bioreactor, EcMEL8 produced 1.06 ± 0.07 g/L NAS and 0.65 ± 0.11 g/L melatonin with tryptophan supplementation.

Conclusions

This study established a novel combinatorial pathway for melatonin biosynthesis in E. coli and provided alternative strategies for improvement of melatonin production.
Knockdown lncRNA CRNDE enhances temozolomide chemosensitivity by regulating autophagy in glioblastoma
Authors: Zhao Zijin , Liu Miaomiao , Long Wenyong , Yuan Jian , Li Haoyu , Zhang Chi , Tang Guodong , Jiang Weixi , Yuan Xianrui , Wu Minghua , Liu Qing
Published: August 2021

Background

The regulatory roles of long non-coding RNA (lncRNA) CRNDE in temozolomide (TMZ) chemoresistance to glioblastoma multiforme (GBM) are still poorly understood. Therefore, the function, characteristics, and possible mechanism of CRNDE in TMZ-induced chemoresistance to GBM were explored.

Methods

Firstly, the expression level of CRNDE in 58 cases of glioma tissue specimens and 30 cases of normal brain tissues were tested by qRT-PCR. Meanwhile, the correlation between CRNDE expression level, the clinicopathological characteristics, and survival time of patients with glioma were analyzed. Then, the CRNDE expression in various glioma cell lines was detected, and CRNDE knockdown cell models were constructed. Subsequently, to explore the effect of CRNDE on chemosensitivity to TMZ, cell viability was detected by the CCK-8 assay and IC50 values, and cell proliferation was detected by cell clone assay and EdU assay, as well as cell survival was detected by apoptosis with flow cytometry under TMZ treatment. Further, the expression of drug-resistance protein ABCG2, autophagy related proteins, and PI3K/Akt/mTOR pathway were measured by western blot or qRT-PCR in TMZ-treated glioma cells. Finally, the mouse tumor xenograft model was established and the tumor volume and weight were measured, and ABCG2 expression was conducted by immunohistochemistry assay.

Results

The integrated results demonstrated lncRNA CRNDE was a poor prognosis factor for GBM patient, which was upregulated in patients who were resistant to TMZ, and closely associated with chemotherapeutic response status to TMZ treatment. Further, functional assays revealed that knockdown of CRNDE could notably reduce glioma cell viability and proliferation, and elevate cell apoptosis to enhance the chemosensitivity to TMZ in vitro and in vivo. Mechanistically, the depression of CRNDE could diminish the expression of LC3 II/I, Beclin1 and Atg5 and increase the p62 expression level to inhibit autophagy due to the activation of PI3K/Akt/mTOR pathway as well as highly correlated with ABCG2 expression.

Conclusions

Overall, the study provided that lncRNA CRNDE is a reliable clinical predictor of outcome and prognosis and a potential biomarker for predicting TMZ treatment response in GBM by modulating the autophagy through PI3K/Akt/mTOR pathway and ABCG2 expression which may be a novel therapeutic target for regulating TMZ sensitivity to GBM.
The role of the deubiquitinating enzyme DUB3/USP17 in cancer: a narrative review
Authors: Yang Guang-Fei , Zhang Xin , Su Yi-Ge , Zhao Ren , Wang Yan-Yang
Published: August 2021

The balance between ubiquitination and deubiquitination is critical for the degradation, transport, localization, and activity of proteins. Deubiquitinating enzymes (DUBs) greatly contribute to the balance of ubiquitination and deubiquitination, and they have been widely studied due to their fundamental role in cancer. DUB3/ubiquitin-specific protease 17 (USP17) is a type of DUB that has attracted much attention in cancer research. In this review, we summarize the biological functions and regulatory mechanisms of USP17 in central nervous system, head and neck, thoracic, breast, gastrointestinal, genitourinary, and gynecologic cancers as well as bone and soft tissue sarcomas, and we provide new insights into how USP17 can be used in the management of cancer.
Lymphocyte activating gene 3 protein expression in nasopharyngeal carcinoma is correlated with programmed cell death-1 and programmed cell death ligand-1, tumor-infiltrating lymphocytes
Authors: Luo Fan , Cao Jiaxin , Lu Feiteng , Zeng Kangmei , Ma Wenjuan , Huang Yan , Zhang Li , Zhao Hongyun
Published: August 2021

Background

Immunotherapy has shown promising efficacy in patients with nasopharyngeal carcinoma (NPC). Lymphocyte activating 3 gene (LAG-3) represents a significant immune target, however, its relationship with NPC remains unclear. This study aimed to evaluate LAG-3 expression in NPC and its association with CD3+ tumor-infiltrating lymphocytes (TILs), Granzyme B (GZMB), programmed death ligand 1 (PD-L1), and programmed death 1 (PD-1) expression.

Methods

A total of 182 patients with NPC from Sun Yat-sen University Cancer Center, China, were included in this retrospective study. LAG-3 expression in 15 NPC cell lines and LAG-3, CD3+ TILs, GZMB, PD-L1 and PD-1 in clinical samples were estimated using immunohistochemistry. The Chi-square test was used to estimate the association between LAG-3, other biomarkers, and clinical characteristics. Survival analysis was performed using the Kaplan–Meier method and the Cox regression model.

Results

LAG-3 was negatively expressed in all of the 15 NPC cell lines, whereas, 147 patients with NPC (80.8%) exhibited high LAG-3 expression on TILs from tumor tissues. Male patients and those who were EBV-positive presented higher LAG-3 expression. Correlation analyses showed that LAG-3 expression was related to PD-1 expression on TILs, as well as, PD-L1 expression on tumor cells (TCs) and TILs. Both the univariate and multivariate Cox models indicated that pathological type III (P = 0.036), higher LAG-3 on TILs (P < 0.001), higher PD-L1 on TCs (P = 0.027), and higher PD-1 on TILs (P < 0.001) were associated with poorer disease-free survival (DFS). However, lower PD-L1 expression on TILs was related to superior DFS only in the univariate Cox analyses (P = 0.002).

Conclusion

Higher LAG-3 and PD-1 on TILs, and higher PD-L1 expression on TCs, and pathological type III were identified as independent risk factors for poorer DFS in NPC patients. Our data demonstrate that LAG-3 is a promising inhibitory receptor that may play an important role in anti-NPC therapy.
Assessing community knowledge, attitude and practices to strengthen communication strategy for Malaria Elimination Demonstration Project in Mandla
Authors: Rajvanshi Harsh , Saha Kalyan B. , Sharma Ravendra K. , Bharti Praveen K. , Nisar Sekh , Jayswar Himanshu , Mishra Ashok K. , Shukla Man Mohan , Das Aparup , Kaur Harpreet , Wattal Suman L. , Lal Altaf A.
Published: August 2021

Background

Changes in social, belief, and behavioural practices are essential for the success of any public health delivery programme. In the planning stages of the Malaria Elimination Demonstration Project (MEDP), priority was given to communication with a goal to develop capacity of health workers and to improve the knowledge, attitude and practices (KAP) of the people of Mandla. This paper describes the level of community knowledge on malaria, including its prevention, diagnosis, treatment-seeking behaviour, and the level of satisfaction with the services provided by the project.

Methods

A cross sectional survey was undertaken in 1233 villages of Mandla to study the KAP and self-assessed improvement in knowledge and satisfaction level of the community. The goal of the study was to understand whether there is need for strengthening communication strategy of MEDP for better impact. The survey was conducted amongst the head/eligible members of the 733 households located in the nine blocks of the district using clustered random sampling.

Results

Though four-fifths of the respondents were able to correlate the transmission of malaria with mosquitoes, misconceptions existed among them. The types of malaria were not known to everyone. Only 39% were aware of the Indoor Residual Spray (IRS) and 41% understood the value of Long-Lasting Insecticidal Nets (LLIN). Around 71% of subjects surveyed were aware of the proper diagnostic tests for malaria. A total of 87% of the respondents knew about the MEDP staff working in their respective villages.

Conclusion

The study reported gaps in knowledge on malaria at community level. The self-assessment of the community revealed that the communication strategy established by MEDP in Mandla district has been useful to them as they are becoming better informed about the prevention and treatment aspects of disease. The lessons learned as revealed in the KAP survey will improve malaria elimination outcomes in a timely manner.
Comparing insecticide-treated nets access-use based on universal household and population indicators vis-a-vis measures adapted to sleeping spaces in Ethiopia
Authors: Kebede Yohannes , Sudhakar Morankar , Alemayehu Guda , Abebe Lakew , Birhanu Zewdie
Published: August 2021

Background

Insecticide-treated nets (ITNs) access-use has been pivotal monitoring indicator for malaria prevention and control, particularly in resource limited settings. The objective of the study was to compare ITN access-use based on universal household and population indicators and measures adapted to sleeping spaces.

Methods

A cross-sectional study was conducted in five districts of Jimma Zone, Ethiopia, March, 2019. 762 HHs were sampled for the survey. Multi-stage followed by simple random sampling used. Monitoring and evaluation reference group’s (MERG’s) indicators were used for measuring ITN access-use. MERG’s indicators are each adapted ITN access-use to sleeping spaces. Household (ownership, saturation and sufficiency) and population access and household members’ status of last night sleeping under ITN compared based on the two models. Differences of estimates of ITN access-use based on the two methods reported as magnitude of over/under estimations, at p-value < 0.05.

Results

Based on MERG’s approach, the study revealed household (HH) based indicators as such: HH ownership of at least 1 ITN (92.6%), sufficiency of ITN for every two people in HH (50.3%), and saturation of ITN for every 2 people in HHs with any ITN (54.6%). Moreover, population based indicators were: population with ITN access (P3 = 78.6%), people who slept under ITN previous night (63.0%), people who slept under ITN among who accessed it (73.1%), ITN use-gap (26.9%). Equivalent indicators of HH ownership, sufficiency, saturation, and people accessed at where they actually slept, and people slept under ITN among those accessed at where they slept estimated at 71.3%, 49.4%, 69.3%, 66.3%, and 92.1%, respectively. MERG’s approach over-estimated ownership, people’s access, and behaviour-failures by 21.3%, 12.3%, 19.0%, respectively. Over-estimation occurred for reasons such as many sleeping spaces lack ITN and > 2 people actually slept per sleeping space.

Conclusions

MERG’s universal indicators over estimated households and populations ITN access-use as a result of absence of measures capturing access-use values at spaces where people actually slept. Consequently, measures adapted to sleeping contexts revealed potential misdistributions practiced when the existing indicators are in use. Insertion of sleeping spaces into existing approach will be worthwhile and needs to be promoted as it improves curiosity in ITN distribution, produces closer estimates and prevents malaria prevention and control programmes from overlooking access-use challenges.
High-resolution probe design for measuring the dielectric properties of human tissues
Authors: Wang Xinran , Guo Hongfu , Zhou Chen , Bai Junkai
Published: August 2021

Background

In order to use the microwave to measure the dielectric constant of the human body and improve the measurement resolution, a small near-field probe working at 915 MHz is designed in this paper.

Method

Based on the electric small loop antenna model loaded by the spiral resonator (SR), a small near-field probe was designed. The probe model is designed and optimized by the HFSS (high frequency structure simulator) software. The human tissues were tested by the manufactured probe and the relationship between the S11 parameters of the probe and the human tissues was analyzed.

Results and conclusions

A probe with small size was designed and fabricated, with the overall size of 10.0 mm × 12.0 mm × 0.8 mm. The probe has a good performance with a 30.7 dB return loss, a 20 MHz bandwidth at the resonance point, and a distance resolution of 10 mm. Due to the small size and good resolution of the probe, it can be used in the measurement of human tissues.