bluuebunny/crossref_metadata_2025 huggingface dataset:
Phenotypic and genetic characterizations of the Milan cohort of von Willebrand disease type 2
Authors: Seidizadeh Omid , Baronciani Luciano , Pagliari Maria Teresa , Cozzi Giovanna , Colpani Paola , Cairo Andrea , Siboni Simona Maria , Biguzzi Eugenia , Peyvandi Flora
Published: April 2022

von Willebrand disease (VWD) type 2 is caused by qualitative abnormalities of von Willebrand factor (VWF). This study aimed to determine the genotypic and phenotypic characterizations of a large VWD type 2 cohort from Milan. We included 321 patients (54% female) within 148 unrelated families from 1995 to 2021. Patients were fully characterized using laboratory phenotypic tests, and the genotypic diagnosis was confirmed by target genetic analysis using Sanger sequencing. Patients were diagnosed with type 2A (n = 98; 48 families), 2B (n = 85; 38 families), 2M (n = 112; 50 families), or 2N (n = 26; 12 families). Eighty-two unique VWF variants, including 8 novel variants, were found. The potential pathogenic effect of novel variants was assessed by in silico analysis. Most patients were heterozygous for a single variant (n = 259; 81%), whereas 37 cases (11%) had 2 variants (4 homozygous, 9 in trans, and 24 in cis). Twenty-five patients (8%) had ≥3 variants, mainly as a result of gene conversions. Among the 82 distinct variants identified, 5 different types, including missense (n = 64), gene conversion (n = 10), synonymous (n = 1), deletion (n = 4), and splice (n = 3), were observed. The results from this large cohort showed that VWD type 2 is invariably due to variants that do not prevent the synthesis of the protein, and a vast majority of patients (88%) had missense variants. Given the complexity of type 2 diagnosis and the necessity of performing several phenotypic tests, genetic analysis for patients suspected of having type 2 is beneficial to establish the correct diagnosis.
Analysis of contributory factors to incidents related to medication assistance for residents taking medicines in residential care homes for the elderly: a qualitative interview survey with care home staff
Authors: Kizaki Hayato , Yamamoto Daisuke , Satoh Hiroki , Masuko Kotaro , Maki Hideyuki , Konishi Yukari , Hori Satoko , Sawada Yasufumi
Published: April 2022

Background

In Japan, staff who are not doctors or nurses can assist the elderly in residential care facilities to take their pre-packaged medicines. Therefore, there is a potential risk of incidents specific to staffs. The aim of this study was to clarify the causes of incidents related to medication assistance by staff in residential care facilities.

Method

Semi-structured interviews with staff involved in medication incidents in long-term care facilities, focusing on how and why each incident happened, were conducted. The interview covered basic information about the subject and resident, the circumstances under which the incident had occurred, contributing factors, and countermeasures put in place. Verbatim transcripts of the interviews were prepared. Based on thematic analysis, codes and themes were created.

Results

Twelve subjects participated in this study. All subjects were staffs (not doctors or nurses) in long-term care facilities. All incidents covered in this study were incidents in which the wrong resident was given the medication. The incidents arose because of “not following procedures”, such as lack of “self-check of residents’ faces/residents’ names/residents’ medicine envelopes” or “double-check with other staff” or “using a device for medication intake”. Contributory factors were grouped into four categories: individual resident factor items such as “decreased ability to understand their medication” or “refusal to take medicines”, individual staff factor items such as “lack of knowledge related to medication” or “mental burden” or “experience in medication assistance”, team factor items such as “failure to communicate with other staff”, work environment factor items such as “presence of other residents” or “other work besides medication assistance” or “not enough time” or “little understanding of fostering a safety culture at the facility”.

Conclusion

This study identified four categories of contributory factors that may lead to incidents during medication assistance by caregivers for residents of care homes. These findings should be helpful for risk management in residential care facilities where staff usually provide medication assistance. Separation of meal times and medication assistance, and professional review to stagger the timing of administration of residents’ medication may be effective in reducing incidents.
Identification and engagement of naturally occurring retirement communities to support healthy aging in Canada: A set of methods for replication
Authors: DePaul Vincent G. , Parniak Simone , Nguyen Paul , Hand Carri , Letts Lori , McGrath Colleen , Richardson Julie , Rudman Debbie , Bayoumi Imaan , Cooper Helen , Tranmer Joan , Donnelly Catherine
Published: April 2022

Background

Naturally occurring retirement communities (NORCs), unplanned communities with a high proportion of older adult residents, offer a model to support older adults to age well in place. The aim of this paper is to provide a comprehensive description of the methods used to identify and engage NORCs appropriate for the development of supportive service programming in Canada.

Methods

Three steps were used to identify and select NORCs in which to develop supportive service programming including: 1) identification of potential NORCs using Canadian Census Dissemination Areas, the Ontario Marginalization Index and Google Maps, 2) engagement of property owner/manager to determine the availability of common space for communal programming and willingness of the owner to support programming and, 3) engagement of older adult residents within the NORC to co-design programming.

Results

Four cities in the south-east, south-central, and south-west of Ontario, Canada were identified to develop NORCs with supportive service programming. Using the methods described, six NORCs were identified, landlords and older adult residents were engaged, and programs initiated between April 2018 and March 2019. The sites included two private high-rise apartments, a city-owned low-rise subsidized apartment complex, two multi-building private high-rise complexes and a mobile home community. An average of 35 (min 20, max 78) older adult members were engaged in an average of 20.5 unique activity sessions at each site per month. On average, social (54%) and physical activities (30%) were more common than nutritional (10%) and knowledge-sharing (8%).

Conclusions

The increased prevalence of unplanned, geographically-bound NORCs creates an opportunity for governments, social and health service providers and policy makers to support healthy aging in their communities. Our experience with the creation of six new NORCs with supportive service programming provides a tested set of methods that can be applied in other communities.
Anti-neutrophil cytoplasmic antibody associated vasculitis in patients with rheumatoid arthritis
Authors: Wu Haiting , Lu Yiyun , Hu Rongrong , Ye Wei , Wen Yubing , Cai Jianfang , Li Hang , Li Xuemei
Published: April 2022

Background

Anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) may coexist with rheumatoid arthritis (RA). However, it is unclear whether the manifestations of AAV with and without coexisting RA are similar. This observational study aimed to investigate the clinicopathological manifestations of AAV with coexisting RA and to explore potential predictors for identifying AAV superimposed on RA.

Methods

Patients with both AAV and RA were identified by searching our hospital database and the literature. Data including age, sex, clinical manifestation, laboratory tests, renal pathology, and therapeutic regimens were retrieved. To assess the difference in clinical features and renal pathology between AAV patients with and without RA, we conducted 1:4 matched case-control studies.

Results

A total of 47 patients were identified, 15 from our hospital and 32 from the literature, and 33 (70.2%) were women. AAV was diagnosed later than RA in 83.0% of the patients and manifested as microscopic polyangiitis (MPA) in 78.7% of the patients. The kidney was the most frequently involved extra-articular organ (74.5%), followed by the lung (51.1%), and skin (8.5%). Patients with both AAV and RA were more likely to be asymptomatic (26.7% vs 3.3%, p = 0.013) than those with isolated AAV. However, they did not differ in other clinicopathological features. In RA patients, those with ANCA associated glomerulonephritis, were more likely to have decreased renal function at renal biopsy as opposed to those with primary glomerulonephritis.

Conclusions

AAV can coexist with RA. In this coexistence, AAV usually develops after RA, is more likely to be asymptomatic, and manifests predominately as MPA with renal involvement. Thus, we should remain vigilant to superimposed AAV on RA.
Impact of Resistance Exercise and Nutritional Endorsement on physical performance in patients with GvHD (IRENE-G study) – design and rational of a randomized controlled trial
Authors: Bujan Rivera Janina , Kühl Rea , Zech Ulrike , Hendricks Anne , Luft Thomas , Dreger Peter , Friedmann-Bette Birgit , Betz Theresa-Maria , Wiskemann Joachim
Published: April 2022

Background

Graft-versus-host disease (GvHD) remains a major complication and limitation to successful allogeneic hematopoietic stem cell transplantation. Treatment of GvHD is challenging due to its heterogeneous nature of presentation, with steroids remaining the established first-line treatment. Long-term doses of systemic corticosteroids have many well-known side-effects including muscle atrophy. Despite the fact that reports in non-cancer clinical populations treated with glucocorticoids demonstrated that resistance training can reverse atrophy and weakness, no RCT has evaluated the potential of resistance training on preventing the disease- and treatment-induced loss of skeletal muscle mass and function in GvHD patients yet. In this context, ensuring adequate nutrition is important as protein deprivation may accelerate the wasting process. As GvHD patients are commonly found to be malnourished, nutritional medical care should be considered when investigating the effect of exercise in GvHD patients. Therefore, the aim of the present “Impact of Resistance Exercise and Nutritional Endorsement on physical performance in patients with GvHD” – Study (IRENE-G) is to evaluate the effects of resistance exercise in combination with nutritional endorsement on physical, nutritional and patient-reported outcomes in GvHD patients.

Methods

IRENE-G is a 24-week prospective interventional RCT. One hundred twelve participants will be randomly allocated (1:1) to one of two arms: resistance exercise and nutritional optimization (experimental) vs. nutritional optimization only (control). Participants in the experimental group will engage in a supervised, progressive moderate-to-high intensity resistance training that is consistent with exercise guidelines for cancer patients, while additionally receiving nutritional support/therapy. Subjects of the control group solely receive nutritional support/therapy based on individual needs. Participants will be assessed at baseline, at 8, 16, 24 weeks for physical performance and various physiological, nutritional and patient-reported outcomes. Follow-up will be 6 months after intervention completion.

Discussion

To our knowledge, this will be the first RCT to assess and compare the effects of a resistance intervention supplemented by nutritional support/therapy against nutritional support only on various health-related outcomes in GvHD patients. The study will contribute to our understanding of the value of exercise and nutritional endorsement in counteracting the negative consequences of GvHD and its treatment.

Trial registration

ClinicalTrials.gov:NCT05111834. Registered 8 November 2021 – Retrospectively registered.
Waist-hip ratio is an independent predictor of moderate-to-severe OSA in nonobese males: a cross-sectional study
Authors: Wang Yan , Mao Lusi , Zhang Xiaolei
Published: April 2022

Background

Adiposity is a well-established risk factor for obstructive sleep apnea (OSA), but whether a combination of preferable anthropometric measurements may improve the accuracy of detecting OSA is unknown. This study aimed to explore the accuracies of the waist-hip ratio (WHR) in conjunction with the body mass index (BMI) when identifying the severity of OSA.

Design

A total of 2012 participants in the China-Japan Friendship Hospital from January 2018 to December 2019 underwent anthropometric measurements and an overnight home sleep test (HST). The 244 subjects who met the criteria for obstructive sleep apnea (apnea–hypopnea index (AHI) ≥ 5 events/hour) were divided into four groups: Group A (55 patients with WHR ≥ 0.9 and BMI ≥ 28 kg/m2); Group B (12 patients with WHR < 0.9 and BMI ≥ 28 kg/m2); Group C (69 patients with WHR ≥ 0.9 and BMI < 28 kg/m2); and group D (108 patients with WHR < 0.9 and BMI < 28 kg/m2).

Results

The AHI, apnea index (AI), hypopnea index (HI), and oxygen desaturation index (ODI) were significantly different among the 4 groups (p < 0.05). The WHR was positively correlated with AHI (r = 0.22, p < 0.001), AI (r = 0.270, p = 0.004), and ODI (r = 0.286, p = 0.0022) and negatively correlated with lowest oxygen pulse saturation (LSpO2) (r = 0.246, p = 0.008) only in nonobese patients. Moreover, the WHR was found to be a screening marker for moderate-to-severe OSA in Group D (p < 0.05). When used to identify severe OSA in Group D, the WHR cut-off point of 0.873 yielded a sensitivity of 65% and specificity of 56% (p < 0.05).

Conclusion

In nonobese male OSA patients, WHR is a moderate screening marker for moderate-to-severe OSA and an independent risk factor for OSA severity.
Only cervical vertebrae C0-C2, not C3 are relevant for subgrouping migraine patients according to manual palpation and pain provocation: secondary analysis of a cohort study
Authors: Schwarz Annika , Luedtke Kerstin , Schöttker-Königer Thomas
Published: April 2022

Background

Subgrouping of migraine patients according to the pain response to manual palpation of the upper cervical spine has been recently described. Based on the neuroanatomy and the convergence of spinal and trigeminal nerves in the trigeminocervical complex, the cervical segments C1 to C3 are potentially relevant. To date it has not been investigated whether palpation results of all upper cervical segments are based on one underlying construct which allows combining the results of several tests. Therefore, the aim of this secondary analysis of a cohort study was to determine whether results from all three segments form one construct.

Methods

Seventy-one migraine patients with chronic or frequent episodic migraine diagnosed according to the international headache society classification version 3 were examined by one physiotherapist. Manual palpation using a posterior to anterior pressure was performed on the upper three cervical vertebrae unilaterally left and right. The results of the palpation according to the patients’ responses were combined using factor analysis. In addition, item response theory (IRT) was used to investigate the structure of the response pattern as well as item difficulty and discrimination.

Findings

Factor analysis (principal component) showed that the palpation of C3 loads less onto the underlying construct than the palpation of C1 and C2. Considering a cut-off value > 1.0, the eigenvalues of all three segments do not represent one underlying construct. When excluding the results from C3, remaining items form one construct. The internal consistency of the pain response to palpation of C1 and C2 is acceptable with a Cronbach’s alpha of 0.69. IRT analysis showed that the rating scale model fits best to the pain response pattern. The discrimination value (1.24) was equal for all items. Item difficulty showed a clear hierarchical structure between the palpation of C1 and C2, indicating that people with a higher impairment are more likely to respond with referred pain during palpation of C2.

Conclusion

Statistical analysis confirms that results from the palpation of the cervical segments C1 and C2 in migraine patients can be combined. IRT analysis confirmed the ordinal pattern of the pain response and showed the higher probability of a pain response during palpation of C2. The pain response to C3 palpation is not relevant for unidimensional IRT analysis.

Trial Registration

German registry of clinical trials (DRKS00015995), Registered 20. December 2018, https://www.drks.de/drks_web/setLocale_EN.do
Palliative care in Uganda: quantitative descriptive study of key palliative care indicators 2018-2020
Authors: Kagarmanova Ainur , Mwesiga Mark Donald , Sisk Matthew L. , Kabagambe Cynthia , Nyakaisiki Sheba , Marentette Tom , Ahern Lacey N.
Published: April 2022

Background

The first and most recent nationwide audit of palliative care services in Uganda was conducted in 2009. Since then, Uganda has made great strides in palliative care development, including policy, education, and services implementation. This study provides an overview of the availability of palliative care services in the country and the challenges and gaps in Uganda prior to the global COVID-19 pandemic. This lays the foundation for better understanding the challenges and changes needed to support palliative care development and access in the wake of the pandemic.

Methods

We conducted a descriptive quantitative study of secondary data on nationwide morphine distribution, collated a list of accredited facilities, and analyzed key palliative care indicators collected through the mHealth surveillance project present at a subset of accredited facilities. Descriptive statistical analysis involved non-parametric tests using SPSS, mapping geographical distribution of available palliative care services using Geographic Information Systems software, and identification of challenges from the subset of accredited facilities.

Results

There were 226 accredited palliative care facilities across Uganda’s 135 districts in 2020. Thirty districts lacked any accredited palliative care facility. The estimated population coverage was 88.5%. The majority (68.1%) of accredited facilities were public, and private facilities received slightly more pain-relieving morphine. There was an alternating trend in the volumes of morphine delivered to public and private facilities. More than a third of the patients were diagnosed with non-communicable diseases, highlighting their significance alongside cancer and HIV/AIDS as conditions requiring palliative care. Palliative care accredited facilities offered six types of services: outreach, home visits, psychosocial, legal, bereavement, and spiritual support, but only for an average of 7 months a year due to lack of facilitation and transportation.

Conclusion

Palliative care in Uganda developed in quality, volume, and geographic coverage since 2009. The shift in palliative care patients’ primary diagnosis from HIV/AIDS to non-communicable diseases marks an important epidemiologic transition. Although accredited facilities are present in most administrative districts, more research is needed to evaluate the actual accessibility of these services. The existing services, both private and public, are limited by the amount of pain-relieving morphine, financial and transport resources. More quality data collected on key palliative care indicators is needed into geographical accessibility of palliative care services, morphine availability trends, and patient diagnoses in order to improve the provision of palliative care in Uganda.
Anti-SARS-CoV-2 potential of Cissampelos pareira L. identified by connectivity map-based analysis and in vitro studies
Authors: Haider Madiha , Anand Vivek , Enayathullah M. Ghalib , Parekh Yash , Ram Sushma , Kumari Surekha , Anmol  , Panda Gayatri , Shukla Manjari , Dholakia Dhwani , Ray Arjun , Bhattacharyya Sudipta , Sharma Upendra , Bokara Kiran Kumar , Prasher Bhavana , Mukerji Mitali
Published: April 2022

Background

Viral infections have a history of abrupt and severe eruptions through the years in the form of pandemics. And yet, definitive therapies or preventive measures are not present. Herbal medicines have been a source of various antiviral compounds such as Oseltamivir, extracted using shikimic acid from star anise (Illicium verum) and Acyclovir fromCarissa edulisare FDA (Food and Drug Administration) approved antiviral drugs. In this study, we dissect the anti-coronavirus infection activity ofCissampelos pareiraL (Cipa) extract using an integrative approach.

Methods

We analysed the signature similarities between predicted antiviral agents andCipausing the connectivity map (https://clue.io/). Next, we tested the anti-SARS-COV-2 activity ofCipain vitro. Molecular docking analyses of constituents of with key targets of SARS-CoV2 protein viz. spike protein, RNA‑dependent RNA‑polymerase (RdRp) and 3C‑like proteinase. was also performed. A three-way comparative analysis ofCipatranscriptome, COVID-19 BALF transcriptome and CMAP signatures of small compounds was also performed.

Results

Several predicted antivirals showed a high positive connectivity score withCipasuch as apcidin, emetine, homoharringtonine etc. We also observed 98% inhibition of SARS-COV-2 replication in infected Vero cell cultures with the whole extract. Some of its prominent pure constituents e.g. pareirarine, cissamine, magnoflorine exhibited 40–80% inhibition. Comparison of genes between BALF andCipashowed an enrichment of biological processes like transcription regulation and response to lipids, to be downregulated inCipawhile being upregulated in COVID-19. CMAP also showed that Triciribine, torin-1 and VU-0365114–2 had positive connectivity with BALF 1 and 2, and negative connectivity withCipa. Amongst all the tested compounds, Magnoflorine and Salutaridine exhibited the most potent and consistent strong in silico binding profiles with SARS-CoV2 therapeutic targets.
How to implement person-centred care and support for dementia in outpatient and home/community settings: Scoping review
Authors: Marulappa Nidhi , Anderson Natalie N. , Bethell Jennifer , Bourbonnais Anne , Kelly Fiona , McMurray Josephine , Rogers Heather L. , Vedel Isabelle , Gagliardi Anna R.
Published: April 2022

Background

Little prior research focused on person-centred care and support (PCCS) for dementia in home, community or outpatient care. We aimed to describe what constitutes PCCS, how to implement it, and considerations for women who comprise the majority of affected persons (with dementia, carers).

Methods

We conducted a scoping review by searching multiple databases from 2000 inclusive to June 7, 2020. We extracted data on study characteristics and PCCS approaches, evaluation, determinants or the impact of strategies to implement PCCS. We used summary statistics to report data and interpreted findings with an existing person-centred care framework.

Results

We included 22 studies with qualitative (55%) or quantitative/multiple methods design (45%) involving affected persons (50%), or healthcare workers (50%). Studies varied in how PCCS was conceptualized; 59% cited a PCC definition or framework. Affected persons and healthcare workers largely agreed on what constitutes PCCS (e.g. foster partnership, promote autonomy, support carers). In 4 studies that evaluated care, barriers of PCCS were reported at the affected person (e.g. family conflict), healthcare worker (e.g. lack of knowledge) and organizational (e.g. resource constraints) levels. Studies that evaluated strategies to implement PCCS approaches were largely targeted to healthcare workers, and showed that in-person inter-professional educational meetings yielded both perceived (e.g. improved engagement of affected persons) and observed (e.g. use of PCCS approaches) beneficial outcomes. Few studies reported results by gender or other intersectional factors, and none revealed if or how to tailor PCCS for women. This synthesis confirmed and elaborated the PCC framework, resulting in a Framework of PCCS for Dementia.

Conclusion

Despite the paucity of research on PCCS for dementia, synthesis of knowledge from diverse studies into a Framework provides interim guidance for those planning or evaluating dementia services in outpatient, home or community settings. Further research is needed to elaborate the Framework, evaluate PCCS for dementia, explore determinants, and develop strategies to implement and scale-up PCCS approaches. Such studies should explore how to tailor PCCS needs and preferences based on input from persons with dementia, and by sex/gender and other intersectional factors such as ethnicity or culture.
Exploring the contexts, mechanisms and outcomes of a torture, abuse and dental anxiety service in Norway: a realist evaluation
Authors: Bryne Emilie , Hean Sarah Catherine Patricia Duff , Evensen Kjersti Berge , Bull Vibeke Hervik
Published: April 2022

Background

Torture, abuse and dental anxiety (TADA) are often precursors to developing a pathological relationship with dental care due to elevated anxiety. Consequently, patients who suffer from one or more of these tend to avoid dental services. This could leave them with severe tooth decay, which could affect their general and psychosocial health. Norwegian dental services have implemented the TADA service to specifically alleviate dental anxiety and restore oral health for the TADA patient group. However, the service has not been evaluated, and there is a need to understand how and why this service works, for whom, under what circumstances. Therefore, this study aimed to develop theories on how the service’s structure alleviates dental anxiety and restores these patients’ oral health. Although developed in a Norwegian context, these theories may be applicable to other national and international contexts.

Methods

This realist evaluation comprised multiple sequential methods of service and policy documents (n = 13), followed by interviews with service developers (n = 12).

Results

The analysis suggests that, by subsidising the TADA service, the Norwegian state has removed financial barriers for patients. This has improved their access to the service and, hence, their service uptake. National guidelines on service delivery are perceived as open to interpretation, and can hereby meet the needs of a heterogeneous patient group. The services have become tailored according to the available regional resources and heterogeneous needs of the patient population. A perceived lack of explicit national leadership and cooperative practices has resulted in regional service teams becoming self-reliant and insular. While this has led to cohesion within each regional service, it is not conducive to interservice collaborations. Lastly, the complexity of migration processes and poor dissemination practices is presumed to be the cause of the lack of recruitment of torture survivors to the service.

Conclusions

Policy documents and service developers described the TADA service as a hybrid bottom-up/top-down service that allows teams to practise discretion and tailor their approach to meet individual needs. Being free of charge has improved access to the service by vulnerable groups, but the service still struggles to reach torture survivors.
Complications and owner satisfaction associated with limb amputation in cats: 59 cases (2007–2017)
Authors: Wagner John R. , DeSandre-Robinson Dana M. , Moore George E. , Loughin Catherine A. , Simons Micha C.
Published: April 2022

Background

Limb amputation may be recommended in domestic cats following a severe injury or disease. The purpose of the study was to report the signalment, the complications, recovery outcome, owner satisfaction and expectations of domestic cats following limb amputation.

Results

Medical records of 3 specialty hospitals were reviewed for cats that received a single limb amputation in a 10 year period (2007–2017). These cat owners were contacted, and 59 owners completed surveys, comprising the study population. The most common reasons for limb amputation were neoplasia (54.2%, 32/59), traumatic injury (40.7%, 24/59), bone or joint infection (3.4%, 2/59), and thromboembolism (1.7%, 1/59). Thirty-four cats (57.6%) had postoperative complications. Of the fifty-nine surveys, 52.5% reported minor complications and 5.1% reported major complications. There were no differences in postoperative complication rates for thoracic versus pelvic limb amputations. All owners reported either excellent (77.9%, 46/59), good (20.3% 12/59), or fair (1.7%, 1/59) satisfaction with the procedure. Based on their previous experiences, 84.7% (50/59) of owners would elect limb amputation if medically warranted for another pet. The remaining 15.3% of owners who would not elect limb amputation again had experienced death of their pet with a median survival time of 183 days.

Conclusion

Owners reported a positive satisfaction when considering complications, recovery outcome, and expectations. This study can be used by veterinarians to guide cat owners in the decision making process of limb amputation.
Synthetic acid stress-tolerance modules improve growth robustness and lysine productivity of industrial Escherichia coli in fermentation at low pH
Authors: Yao Xurong , Liu Peng , Chen Bo , Wang Xiaoyan , Tao Fei , Lin Zhanglin , Yang Xiaofeng
Published: April 2022

Background

During fermentation, industrial microorganisms encounter multiple stresses that inhibit cell growth and decrease fermentation yields, in particular acid stress, which is due to the accumulation of acidic metabolites in the fermentation medium. Although the addition of a base to the medium can counteract the effect of acid accumulation, the engineering of acid-tolerant strains is considered a more intelligent and cost-effective solution. While synthetic biology theoretically provides a novel approach for devising such tolerance modules, in practice it is difficult to assemble stress-tolerance modules from hundreds of stress-related genes.

Results

In this study, we designed a set of synthetic acid-tolerance modules for fine-tuning the expression of multi-component gene blocks comprising a member of the proton-consuming acid resistance system (gadE), a periplasmic chaperone (hdeB), and reactive oxygen species (ROS) scavengers (sodB and katE). Directed evolution was used to construct an acid-responsive asr promoter library, from which four variants were selected and used in the synthetic modules. The module variants were screened in a stepwise manner under mild acidic conditions (pH 5–6), first by cell growth using the laboratory Escherichia coli strain MG1655 cultured in microplates, and then by lysine production performance using the industrial lysine-producing E. coli strain MG1655 SCEcL3 cultured first in multiple 10-mL micro-bioreactors, and then in 1.3-L parallel bioreactors. The procedure resulted in the identification of a best strain with lysine titer and yield at pH 6.0 comparable to the parent strain at pH 6.8.

Conclusion

Our results demonstrate a promising synthetic-biology strategy to enhance the growth robustness and productivity of E. coli upon the mildly acidic conditions, in both a general lab strain MG1655 and an industrial lysine-producing strain SCEcL3, by using the stress-responsive synthetic acid-tolerance modules comprising a limited number of genes. This study provides a reliable and efficient method for achieving synthetic modules of interest, particularly in improving the robustness and productivity of industrial strains.
A single point mutation in the Plasmodium falciparum 3′–5′ exonuclease does not alter piperaquine susceptibility
Authors: Boonyalai Nonlawat , Kirativanich Kirakarn , Thamnurak Chatchadaporn , Praditpol Chantida , Vesely Brian A. , Wojnarski Mariusz , Griesenbeck John S. , Waters Norman C.
Published: April 2022

Background

The rise in Plasmodium falciparum resistance to dihydroartemisinin–piperaquine (DHA–PPQ) treatment has been documented in the Greater Mekong Subregion with associations with mutations in the P. falciparum chloroquine resistance transporter (pfcrt) and plasmepsin 2 (pfpm2) genes. However, it is unclear whether other genes also play a role with PPQ resistance, such as the E415G mutation in the exonuclease (pfexo) gene. The aim of this study was to investigate the role of this mutation in PPQ resistance by generating transgenic parasites expressing the pfexo-E415G mutant allele.

Methods

Transgenic parasite clones carrying the E415G mutation in PfEXO of the B5 isolate were derived by CRISPR-Cas9 gene editing and verified using PCR and gene sequencing. Polymorphisms of pfkelch-13, pfcrt, and pfexo were examined by PCR while the copy number variations of pfpm2 were examined by both relative quantitative real-time PCR and the duplication breakpoint assay. Drug sensitivity against a panel of antimalarials, the ring-stage survival assay (RSA), the PPQ survival assay (PSA), and bimodal dose-response curves were used to evaluate antimalarial susceptibility.

Results

The transgenic line, B5-rexo-E415G-B8, was successfully generated. The PPQ-IC90, %PPQ survival, and the bimodal dose-response clearly showed that E415G mutation in PfEXO of B5 isolate remained fully susceptible to PPQ. Furthermore, growth assays demonstrated that the engineered parasites grew slightly faster than the unmodified parental isolates whereas P. falciparum isolates harbouring pfkelch-13, pfcrt, and pfexo mutations with multiple copies of pfpm2 grew much more slowly.

Conclusions

Insertion of the E415G mutation in PfEXO did not lead to increased PPQ-IC90 and %PPQ survival, suggesting that this mutation alone may not be associated with PPQ resistance, but could still be an important marker if used in conjunction with other markers for monitoring PPQ-resistant parasites. The results also highlight the importance of monitoring and evaluating suspected genetic mutations with regard to parasite fitness and resistance.
CDK6-PI3K signaling axis is an efficient target for attenuating ABCB1/P-gp mediated multi-drug resistance (MDR) in cancer cells
Authors: Zhang Lei , Li Yidong , Hu Chaohua , Chen Yangmin , Chen Zhuo , Chen Zhe-Sheng , Zhang Jian-Ye , Fang Shuo
Published: April 2022

Background

Multidrug resistance (MDR) mediated by ATP binding cassette subfamily B member 1 (ABCB1/P-gp) is a major cause of cancer chemotherapy failure, but the regulation mechanisms are largely unknown.

Methods

Based on single gene knockout, we studied the regulation of CDK6-PI3K axis on ABCB1-mediated MDR in human cancer cells. CRISPR/Cas9 technique was performed in KB-C2 cells to knockout cdk6 or cdk4 gene. Western blot, RT-PCR and transcriptome analysis were performed to investigate target gene deletion and expression of critical signaling factors. The effect of cdk4 or cdk6 deficiency on cell apoptosis and the cell cycle was analyzed using flow cytometry. In vivo studies were performed to study the sensitivity of KB-C2 tumors to doxorubicin, tumor growth and metastasis.

Results

Deficiency of cdk6 led to remarkable downregulation of ABCB1 expression and reversal of ABCB1-mediated MDR. Transcriptomic analysis revealed that CDK6 knockout regulated a series of signaling factors, among them, PI3K 110α and 110β, KRAS and MAPK10 were downregulated, and FOS-promoting cell autophagy and CXCL1-regulating multiple factors were upregulated. Notably, PI3K 110α/110β deficiency in-return downregulated CDK6 and the CDK6-PI3K axis synergizes in regulating ABCB1 expression, which strengthened the regulation of ABCB1 over single regulation by either CDK6 or PI3K 110α/110β. High frequency of alternative splicing (AS) of premature ABCB1 mRNA induced by CDK6, CDK4 or PI3K 110α/110β level change was confirmed to alter the ABCB1 level, among them 10 common skipped exon (SE) events were found. In vivo experiments demonstrated that loss of cdk6 remarkably increased the sensitivity of KB-C2 tumors to doxorubicin by increasing drug accumulation of the tumors, resulting in remarkable inhibition of tumor growth and metastasis, as well as KB-C2 survival in the nude mice.

Conclusions

CDK6-PI3K as a new target signaling axis to reverse ABCB1-mediated MDR is reported for the first time in cancers. Pathways leading to inhibition of cancer cell proliferation were revealed to be accompanied by CDK6 deficiency.
Clinicopathological characteristics and outcomes of malignant adenomyoepithelioma of the breast: a single institution’s experience
Authors: Alqudaihi Heba Mohammad Abdulla , Lee Sae Byul , Son Byung Ho , Ahn Sei-Hyun , Lee Jong Won , Ko Beom Seok , Kim Hee Jeong , Chung Il Yong , Kim Jisun , Gong Gyungyub
Published: April 2022

Background

Malignant adenomyoepithelioma of the breast is a rare tumor and most of relevant literature consists of individual case reports. This study objective was designed to evaluate clinicopathological features and treatment outcomes of 15 cases of malignant adenomyoepithelioma at a single institute.

Methods

A retrospective medical record review was performed for 15 subjects confirmed with malignant adenomyoepithelioma upon postoperative pathological diagnosis at the Asan Medical Center from January 2008 to June 2018. Data regarding age at diagnosis, preoperative biopsy results, operation methods, the status of hormone receptors and HER2, and clinical outcomes were collected.

Results

All cases were female patients diagnosed at median age of 50 years. Preoperative core needle biopsy results showed that 40% of the cases (6 out of 15) were benign which was in discordance with the final malignant pathology report. Thirteen cases underwent wide excision with or without sentinel lymph node biopsy (SLNB) and 2 cases had total mastectomy with SLNB. Five of 11 cases (45.5%) were triple negative. Ten of 15 cases underwent postoperative radiation therapy, 3 cases underwent chemotherapy, and 5 cases underwent endocrine therapy. During median follow-up of 55 months, the 5-year overall survival rate was 87.5% and the 5-year disease free survival rate was 91.7%. Two lung metastases developed. One case showed local recurrence 3 years after surgery and radiotherapy and subsequently developed lung metastasis 1 year late. Another case developed lung metastasis one and a half years after surgery in combination with endocrine therapy and neoadjuvant chemotherapy.

Conclusion

Preoperative core needle biopsy showed inaccurate results for diagnosing malignant adenomyoepithelioma. Malignant adenomyoepithelioma has a high rate of triple negative subtype but has a relatively good prognosis although there is a risk of local and systemic recurrence.
Assessing differences in contraceptive provision through telemedicine among reproductive health providers during the COVID-19 pandemic in the United States
Authors: Comfort Alison B. , Rao Lavanya , Goodman Suzan , Raine-Bennett Tina , Barney Angela , Mengesha Biftu , Harper Cynthia C.
Published: April 2022

Background

Providers faced challenges in maintaining patient access to contraceptive services and public health safety during the COVID-19 pandemic. Due to increased barriers to care, providers increasingly used telemedicine for contraceptive care, curbside services, mail-order pharmacies, and on-line or home delivery of contraceptive methods, including self-administration of subcutaneous depo medroxyprogesterone acetate (DMPA-SQ). To better understand how reproductive health providers adapted service provision during the pandemic, this study assessed clinical practice changes and strategies providers adopted throughout the United States to maintain contraceptive care, particularly when clinics closed on-site, and the challenges that remained in offering contraceptive services, especially to marginalized patient populations.

Methods

We surveyed U.S. providers and clinic staff (n = 907) in April 2020–January 2021, collecting data on contraceptive service delivery challenges and adaptations, including telemedicine. We assessed clinical practice changes with multivariate regression analyses using generalized linear models with a Poisson distribution and cluster robust standard errors, adjusting for clinic patient volume, practice setting, region, Title X funding, and time of survey.

Results

While 80% of providers reported their clinic remained open, 20% were closed on-site. Providers said the pandemic made it more difficult to offer the full range of contraceptive methods (65%), contraceptive counseling (61%) or to meet the needs of patients in marginalized communities (50%). While only 11% of providers offered telemedicine pre-pandemic, most offered telemedicine visits (79%) during the pandemic. Some used mail-order pharmacies (35%), curbside contraceptive services (22%), and DMPA-SQ for self-administration (10%). Clinics that closed on-site were more likely to use mail-order pharmacies (aRR 1.83, 95% CI [1.37–2.44]) and prescribe self-administered DMPA-SQ (aRR 3.85, 95% CI [2.40–6.18]). Clinics closed on-site were just as likely to use telemedicine as those that remained open. Among clinics using telemedicine, those closed on-site continued facing challenges in contraceptive service provision.

Conclusions

Clinics closing on-site were just as likely to offer telemedicine, but faced greater challenges in offering contraceptive counseling and the full range of contraceptive methods, and meeting the needs of marginalized communities. Maintaining in-person care for contraceptive services, in spite of staffing shortages and financial difficulties, is an important objective during and beyond the pandemic.
Altered gene expression of VEGF, IGFs and H19 lncRNA and epigenetic profile of H19-DMR region in endometrial tissues of women with endometriosis
Authors: Kamrani Sedigheh , Amirchaghmaghi Elham , Ghaffari Firouzeh , Shahhoseini Maryam , Ghaedi Kamran
Published: April 2022

Background

Endometriosis, as chronic estrogen-dependent disease, is defined by the presence of endometrial-like tissue outside the uterus. Proliferation of endometrial tissue and neoangiogenesis are critical factors in development of endometriosis. Hence, vascular endothelial growth factor (VEGF) as well as insulin‐like growth factor 1 and 2 (IGF1, 2) may be involved as inducers of cellular proliferation or neoangiogenesis. Imprinted long noncoding RNA H19 (lncRNA H19) has been suggested to be involved in pathogenesis of endometriosis via regulation of cellular proliferation and differentiation. Epigenetic aberrations appear to play an important role in its pathogenesis. The present study was designed to elucidate VEGF, IGF1, IGF2 and H19 lncRNA genes expression and epigenetic alterations of differentially methylated region (DMR) of H19 (H19-DMR) regulatory region in endometrial tissues of patients with endometriosis, in comparison with control women.

Methods

In this case–control study, 24 women with and without endometriosis were studied for the relative expression of VEGF, IGF1, IGF2 and H19 lncRNA genes using real-time polymerase chain reaction (PCR) technique. Occupancy of the MeCP2 on DMR region of H19 gene was assessed using chromatin immunoprecipitation (ChIP), followed by real-time PCR.

Results

Genes expression profile of H19, IGF1 and IGF2 was decreased in eutopic and ectopic endometrial tissues of endometriosis group, compared to the control tissues. Decreased expression of H19 in ectopic samples was significant in comparison with the controls (P < 0.05). Gene expression of VEGF was increased in eutopic tissues of endometriosis group, compared to control group. Whereas its expression level was lower in ectopic lesions versus eutopic and control endometrial samples. ChIP analysis revealed significant and nearly significant hypomethylation of H19-DMR region II in eutopic and ectopic samples, compared to the control group respectively. This epigenetic change was aligned with expression of IGF2. While methylation of H19-DMR region I was not significantly different between the eutopic, ectopic and control endometrial samples.

Conclusion

These data showed that VEGF, IGF1, IGF2 and H19 lncRNA genes expression and epigenetic alterations of H19 lncRNA have dynamic role in the pathogenesis of endometriosis, specifically in the way that hypomethylation of H19-DMR region II can be involved in IGF2 dysregulation in endometriosis.
Settings matter: a scoping review on parameters in robot-assisted gait therapy identifies the importance of reporting standards
Authors: van Dellen Florian , Labruyère Rob
Published: April 2022

Background

Lokomat therapy for gait rehabilitation has become increasingly popular. Most evidence suggests that Lokomat therapy is equally effective as but not superior to standard therapy approaches. One reason might be that the Lokomat parameters to personalize therapy, such as gait speed, body weight support and Guidance Force, are not optimally used. However, there is little evidence available about the influence of Lokomat parameters on the effectiveness of the therapy. Nevertheless, an appropriate reporting of the applied therapy parameters is key to the successful clinical transfer of study results. The aim of this scoping review was therefore to evaluate how the currently available clinical studies report Lokomat parameter settings and map the current literature on Lokomat therapy parameters.

Methods and results

A systematic literature search was performed in three databases: Pubmed, Scopus and Embase. All primary research articles performing therapy with the Lokomat in neurologic populations in English or German were included. The quality of reporting of all clinical studies was assessed with a framework developed for this particular purpose. We identified 208 studies investigating Lokomat therapy in patients with neurologic diseases. The reporting quality was generally poor. Less than a third of the studies indicate which parameter settings have been applied. The usability of the reporting for a clinical transfer of promising results is therefore limited.

Conclusion

Although the currently available evidence on Lokomat parameters suggests that therapy parameters might have an influence on the effectiveness, there is currently not enough evidence available to provide detailed recommendations. Nevertheless, clinicians should pay close attention to the reported therapy parameters when translating research findings to their own clinical practice. To this end, we propose that the quality of reporting should be improved and we provide a reporting framework for authors as a quality control before submitting a Lokomat-related article.