bluuebunny/crossref_metadata_2025 huggingface dataset:
Involvement of the Cell Wall Integrity Pathway of Saccharomyces cerevisiae in Protection against Cadmium and Arsenate Stresses
Authors: Techo Todsapol , Charoenpuntaweesin Sirada , Auesukaree Choowong
Published: August 2020

Environmental pollution by metal/metalloids such as cadmium and arsenic has become a serious problem in many countries, especially in developing countries. This study shows that in the yeast
S. cerevisiae
, the CWI pathway plays a protective role against cadmium and arsenate through the upregulation of genes involved in cell wall biosynthesis and cell cycle control, possibly in order to modulate cell wall reconstruction and cell cycle phase transition, respectively. These data provide insights into molecular mechanisms underlying adaptive responses to cadmium and arsenate.
Virulence Genetics of anErwinia amylovoraPutative Polysaccharide Transporter Family Member
Authors: Klee Sara M. , Sinn Judith P. , Christian Elena , Holmes Aleah C. , Zhao Kaixi , Lehman Brian L. , Peter Kari A. , Rosa Cristina , McNellis Timothy W.
Published: August 2020

The Gram-negative enterobacteriumErwinia amylovoracauses fire blight disease in apple and pear trees. Lipopolysaccharides and the exopolysaccharide amylovoran are essentialE. amylovoravirulence factors. We found that mutations inrfbXdisrupted amylovoran production and virulence in apple fruits and tree shoots and that the deletion ofyibDsuppressed therfbXmutant phenotype. The level of expression ofyibDwas about 10-fold higher in the ΔrfbXmutant than the wild type. A forward genetic suppressor screen in the ΔrfbXmutant uncovered multiple mutations inyibDand supported the conclusion that the virulence defect ofrfbXmutants is due to reduced amylovoran production. TheyibDandrfbXgenes are expressed as a two-gene operon,yibD rfbX. TherfbXgene encodes a previously uncharacterized putative polysaccharide subunit transporter, whileyibDencodes a predicted glycosyltransferase. Mutation ofrfbXdid not have a detectable effect on lipopolysaccharide patterns; however, the overexpression ofyibDin both the wild-type and ΔyibDΔrfbXgenetic backgrounds disrupted both amylovoran and lipopolysaccharide production. Additionally, the overexpression ofyibDin the ΔyibDΔrfbXmutant inhibited bacterial growth in amylovoran-inducing medium. This growth inhibition phenotype was used in a forward genetic suppressor screen and reverse-genetics tests to identify several genes involved in lipopolysaccharide production, which, when mutated, restored the ability of the ΔyibDΔrfbXmutant overexpressingyibDto grow in amylovoran-inducing medium. Remarkably, all the lipopolysaccharide gene mutants tested were defective in lipopolysaccharide and amylovoran production. These results reveal a genetic connection between amylovoran and lipopolysaccharide production inE. amylovora.

IMPORTANCEThis study discovered previously unknown genetic connections between exopolysaccharide and lipopolysaccharide production in the fire blight pathogenErwinia amylovora. This represents a step forward in our understanding of the biology underlying the production of these two macromolecules. Fire blight is an economically important disease that impacts the production of apples and pears worldwide. Few fire blight control measures are available, and growers rely heavily on antibiotic applications at bloom time. Both exopolysaccharide and lipopolysaccharide areE. amylovoravirulence factors. Our results indicate that the overexpression of theyibDgene inE. amylovoradisrupts both lipopolysaccharide production and exopolysaccharide production. This effect could potentially be used as the basis for the development of an antivirulence treatment for the prevention of fire blight disease.
Airway Delivery of Anti-influenza Monoclonal Antibodies Results in Enhanced Antiviral Activities and Enables Broad-Coverage Combination Therapies
Authors: Vigil Adam , Frias-Staheli Natalia , Carabeo Teresa , Wittekind Michael
Published: August 2020

Influenza causes widespread illness in humans and can result in morbidity and death, especially in the very young and elderly populations. Because influenza vaccination can be poorly effective some years, and the immune systems of the most susceptible populations are often compromised, passive immunization treatments using broadly neutralizing antibodies is a promising therapeutic approach. However, large amounts of a single antibody are required for effectiveness when delivered through systemic administration (typically intravenous infusion), precluding the feasible dosing of antibody combinations via this route. The significance of our research is the demonstration that effective therapeutic treatments of multiple relevant influenza types (H1N1, H3N2, and B) can be achieved by airway administration of a single combination of relatively small amounts of three anti-influenza antibodies. This advance exploits the discovery that airway delivery is a more potent way of administering anti-influenza antibodies compared to systemic delivery, making this a feasible and cost-effective therapeutic approach.
The Dynamic Interface of Viruses with STATs
Authors: Harrison Angela R. , Moseley Gregory W.
Published: August 2020

Viruses commonly antagonize the antiviral type I interferon response by targeting signal transducer and activator of transcription 1 (STAT1) and STAT2, key mediators of interferon signaling. Other STAT family members mediate signaling by diverse cytokines important to infection, but their relationship with viruses is more complex. Importantly, virus-STAT interaction can be antagonistic or stimulatory depending on diverse viral and cellular factors. While STAT antagonism can suppress immune pathways, many viruses promote activation of specific STATs to support viral gene expression and/or produce cellular conditions conducive to infection.
Development of a Fluorescence-Based, High-Throughput SARS-CoV-2 3CL
pro
Reporter Assay
Authors: Froggatt Heather M. , Heaton Brook E. , Heaton Nicholas S.
Published: August 2020

The COVID-19 pandemic has already led to more than 700,000 deaths and innumerable changes to daily life worldwide. Along with development of a vaccine, identification of effective antivirals to treat infected patients is of the highest importance. However, rapid drug discovery requires efficient methods to identify novel compounds that can inhibit the virus. In this work, we present a method for identifying inhibitors of the SARS-CoV-2 main protease, 3CL
pro
. This reporter-based assay allows for antiviral drug screening in human cell culture at biosafety level 2 (BSL2) with high-throughput compatible sample processing and analysis. This assay may help identify novel antivirals to control the COVID-19 pandemic.
Ubiquitin Modification of the Epstein-Barr Virus Immediate Early Transactivator Zta
Authors: Zhao Mengmeng , Nanbo Asuka , Becnel David , Qin Zhiqiang , Morris Gilbert F. , Li Li , Lin Zhen
Published: August 2020

Epstein-Barr virus (EBV) is a ubiquitous human pathogen and associated with various human diseases. EBV undergoes latency and lytic replication stages in its life cycle. The transition into the lytic replication stage, at which virus is produced, is mainly regulated by the viral gene product, Zta. Therefore, the regulation of Zta function becomes a central issue regarding viral biology and pathogenesis. Known modifications of Zta include phosphorylation and sumoylation. Here, we report the role of ubiquitination in regulating Zta function. We found that Zta is subjected to ubiquitination in both EBV-infected and EBV-negative cells. The ubiquitin modification targets 4 lysine residues on Zta, leading to both mono- and polyubiquitination of Zta. Ubiquitination of Zta affects the protein’s stability and likely contributes to the progression of viral lytic replication. The function and fate of Zta may be determined by the specific lysine residue being modified.
Application of a Biologically Contained Reporter System To Study Gain-of-Function H5N1 Influenza A Viruses with Pandemic Potential
Authors: Spieler Eva E. , Moritz Eva , Stertz Silke , Hale Benjamin G.
Published: August 2020

Understanding how animal influenza viruses can adapt to spread in humans is critical to prepare for, and prevent, new pandemics. However, working safely with pathogens that have pandemic potential requires tight regulation and the use of high-level physical and biological risk mitigation strategies to stop accidental loss of containment. Here, we used a biological containment system for influenza viruses to study strains with pandemic potential. The system relies on deletion of the essential HA gene from the viral genome and its provision by a genetically modified cell line, to which virus propagation is therefore restricted. We show that this method permits safe handling of these pathogens, including gain-of-function variants, without the risk of generating fully infectious viruses. Furthermore, we demonstrate that this system can be used to assess virus sensitivity to both approved and experimental drugs, as well as the antigenic profile of viruses, important considerations for evaluating prepandemic vaccine and antiviral strategies.
The Antifungal Protein AfpB Induces Regulated Cell Death in Its Parental Fungus
Penicillium digitatum
Authors: Bugeda Adrià , Garrigues Sandra , Gandía Mónica , Manzanares Paloma , Marcos Jose F. , Coca María
Published: August 2020

Disease-causing fungi pose a serious threat to human health and food safety and security. The limited number of licensed antifungals, together with the emergence of pathogenic fungi with multiple resistance to available antifungals, represents a serious challenge for medicine and agriculture. Therefore, there is an urgent need for new compounds with high fungal specificity and novel antifungal mechanisms. Antifungal proteins in general, and AfpB from
Penicillium digitatum
in particular, are promising molecules for the development of novel antifungals. This study on AfpB’s mode of action demonstrates its potent, specific fungicidal activity through the interaction with multiple targets, presumably reducing the risk of evolving fungal resistance, and through a regulated cell death process, uncovering this protein as an excellent candidate for a novel biofungicide. The in-depth knowledge on AfpB mechanistic function presented in this work is important to guide its possible future clinical and agricultural applications.
Antibiotic Resistance in Vibrio cholerae: Mechanistic Insights from IncC Plasmid-Mediated Dissemination of a Novel Family of Genomic Islands Inserted at
trmE
Authors: Rivard Nicolas , Colwell Rita R. , Burrus Vincent
Published: August 2020

The increasing association of the etiological agent of cholera,
Vibrio cholerae
serogroup O1 and O139, with multiple antibiotic resistance threatens to deprive health practitioners of this effective tool. Drug resistance in cholera results mainly from acquisition of mobile genetic elements. Genomic islands conferring multidrug resistance and mobilizable by IncC conjugative plasmids were reported to circulate in non-O1/non-O139
V. cholerae
clinical strains isolated from the 2010 Haitian cholera outbreak. As these genomic islands can be transmitted to pandemic
V. cholerae
serogroups, their mechanism of transmission needed to be investigated. Our research revealed plasmid- and genomic island-encoded factors required for the resistance island excision, mobilization, and integration, as well as regulation of these functions. The discovery of related genomic islands carrying diverse phage resistance genes but lacking antibiotic resistance-conferring genes in a wide range of marine dwelling bacteria suggests that these elements are ancient and recently acquired drug resistance genes.
A Ceftazidime-Avibactam-Resistant and Carbapenem-Susceptible Klebsiella pneumoniae Strain Harboring
bla
KPC-14
Isolated in New York City
Authors: Niu Siqiang , Chavda Kalyan D. , Wei Jie , Zou Chunhong , Marshall Steven H. , Dhawan Puneet , Wang Deqiang , Bonomo Robert A. , Kreiswirth Barry N. , Chen Liang
Published: August 2020

KPC is currently the most common carbapenemase identified in the United States. More than 40 KPC variants have been described, of which KPC-2 and KPC-3 are the most frequent clinical variants. However, our understanding of the genetic structures and β-lactam resistance profiles of other novel KPC variants remains incomplete. Here, we report a novel
bla
KPC
variant (
bla
KPC-14
) and the complete genome sequence of
bla
KPC-14
-harboring
K. pneumoniae
strain BK13048, which is susceptible to carbapenems but resistant to ceftazidime-avibactam. To the best of our knowledge, this is one of the earliest KPC-producing
K. pneumoniae
strains exhibiting resistance to ceftazidime-avibactam.
Development of a Plasmid Shuttle Vector System for Genetic Manipulation of Chlamydia psittaci
Authors: Shima Kensuke , Weber Mary M. , Schnee Christiane , Sachse Konrad , Käding Nadja , Klinger Matthias , Rupp Jan
Published: August 2020

Psittacosis, caused by avian
C. psittaci
, has a major economic impact in the poultry industry worldwide and represents a significant risk for zoonotic transmission to humans. In the past decade, the tools of genetic manipulation have been improved for chlamydial molecular studies. While several genetic tools have been mainly developed in
Chlamydia trachomatis
, a stable gene-inducible shuttle vector system has not to date been available for
C. psittaci
. In this study, we adapted a
C. trachomatis
plasmid shuttle vector system to
C. psittaci
. We constructed a
C. psittaci
plasmid backbone shuttle vector called pCps-Tet-mCherry. The construct expresses GFP in
C. psittaci
. Importantly, exogeneous genes can be inserted at an MCS and are regulated by a tet promoter. The application of the pCps-Tet-mCherry shuttle vector system enables a promising new approach to investigate unknown gene functions of this pathogen.
Stagnant slab front within the mantle transition zone controls the formation of Cenozoic intracontinental high-Mg andesites in northeast Asia
Authors: Xu Wen-Liang , Chen Jia-Hui , Weng Ai-Hua , Tang Jie , Wang Feng , Wang Chun-Guang , Guo Peng , Wang Yi-Ni , Yang Hao , Sorokin Andrey A.
Published: August 2020

The geochemistry of Cenozoic intracontinental high-Mg andesites (HMAs) in northeast Asia, together with regional geophysical data, offers an opportunity to explore the genetic relationship between the formation of intracontinental HMAs and subduction of the Pacific plate. Compared with primary HMAs in arcs, Cenozoic intracontinental HMAs in northeast Asia have lower Mg# [100 × Mg/(Mg + Fe2+)] values (53–56) and CaO contents (5.8–6.6 wt%), higher alkali (Na2O + K2O) contents (5.15–6.45 wt%), and enriched Sr-Nd-Hf isotopic compositions (87Sr/86Sr = 0.7056–0.7059; εNd = −4.9 to −3.4; εHf = −4.7 to −2.6) as well as lower Pb isotope ratios (206Pb/204Pb = 16.76–19.19; 207Pb/204Pb = 15.42–15.45; 208Pb/204Pb = 36.71–37.11). These Cenozoic intracontinental HMAs are similar to Cenozoic potassic basalts in northeast China with respect to their Sr-Nd-Pb-Hf isotopic compositions but have higher SiO2 and Al2O3 contents and lower K2O, MgO, and light rare earth element contents. These features indicate that these Cenozoic intracontinental HMAs originated from the mantle, where recycled ancient sediments and water contributed to partial melting of peridotite. Combined with the presence of a large low-resistivity anomaly derived from the mantle transition zone (MTZ) near these intracontinental HMAs, and their occurrence above the stagnant slab front within the MTZ (at 600 km depth) in northeast Asia, we conclude that the stagnant slab front, with high contents of recycled ancient sediments and water, has controlled the formation of Cenozoic intracontinental HMAs in northeast Asia.
Impact of fault damage on eastern Tibet topography
Authors: Kirkpatrick Heather M. , Moon Seulgi , Yin An , Harrison T. Mark
Published: August 2020

Tectonic deformation can influence spatiotemporal patterns of erosion by changing both base level and the mechanical state of bedrock. Although base-level change and the resulting erosion are well understood, the impact of tectonic damage on bedrock erodibility has rarely been quantified. Eastern Tibet, a tectonically active region with diverse lithologies and multiple active fault zones, provides a suitable field site to understand how tectonic deformation controls erosion and topography. In this study, we quantified erosion coefficients using the relationship between millennial erosion rates and the corresponding channel steepness. Our work shows a twofold increase in erosion coefficients between basins within 15 km of major faults compared to those beyond 15 km, suggesting that tectonic deformation through seismic shaking and rock damage significantly affects eastern Tibet erosion and topography. This work demonstrates a field-based, quantitative relationship between rock erodibility and fault damage, which has important implications for improving landscape evolution models.
Coronoidoplasty in TMJ ankylosis treatment
Authors: Selvaraj Daniel Sathiya Sundaram , Ommen Ajish George , Ebenezer Jagadish
Published: August 2020

A 2-year-old boy was brought by his parents with complaints of difficulty in mouth opening for the past one and half years. He had difficulty in chewing and was malnourished, with developing facial asymmetry. He was diagnosed with right side temporomandibular joint ankylosis. We planned for surgical removal of the ankylotic mass. But we modified the treatment protocol. Instead of doing coronoidectomy after aggressive excision of the ankylotic mass as advocated by Kaban, we did a ‘coronoidoplasty’ after aggressive excision of the ankylotic mass. Coronoidotomy or coronoidectomy is one of the rungs in the treatment ladder that is followed in surgical management of temporomandibular joint ankylosis. But one of the postoperative complications after coronoidectomy is the open bite. The difficulty to close the mouth becomes more pronounced when bilateral coronoidectomy is done. However, ‘coronoidoplasty’, as we have done for this patient retains the action of the temporalis muscle on the mandible in closing the mouth, yet removes the mechanical interference of the coronoid process. Postoperatively the patient was able to clench his teeth well, chew properly and there was no open bite.
Rebalancing glucolipid metabolism and gut microbiome dysbiosis by nitrate-dependent alleviation of high-fat diet-induced obesity
Authors: Ma Linsha , Hu Liang , Jin Luyuan , Wang Jiangyi , Li Xiangchun , Wang Weili , Chang Shimin , Zhang Chunmei , Wang Jingsong , Wang Songlin
Published: August 2020

Introduction

High-fat diet (HFD)-induced obesity is accompanied by compromised nitric oxide (NO) signaling and gut microbiome dysregulation. Inorganic dietary nitrate, which acts as a NO donor, exerts beneficial effects on metabolic disorders. Here, we evaluated the effects of dietary nitrate on HFD-induced obesity and provided insights into the underlying mechanism.

Research design and methods

To investigate the preventive effect of dietary nitrate on HFD-induced obesity, C57BL/6 mice were randomly assigned into four groups (n=10/group), including normal control diet group (normal water and chow diet), HFD group (normal water and HFD), HFD+NaNO3group (water containing 2 mM NaNO3and HFD), and HFD+NaCl group (water containing 2 mM NaCl and HFD). During the experiment, body weight was monitored and glucolipid metabolism was evaluated. The mechanism underlying the effects of nitrate on HFD-induced obesity was investigated by the following: the NO3--NO2--NO pathway; endothelial NO synthase (eNOS) and cyclic guanosine monophosphate (cGMP) levels; gut microbiota via 16SRNA analysis.

Results

Dietary nitrate reduced the body weight gain and lipid accumulation in adipose and liver tissues in HFD-fed mice. Hyperlipidemia and insulin resistance caused by HFD were improved in mice supplemented with nitrate. The level of eNOS was upregulated by nitrate in the serum, liver, and inguinal adipose tissue. Nitrate, nitrite, and cGMP levels were decreased in mice fed on HFD but reversed in the HFD+NaNO3group. Nitrate also rebalanced the colon microbiota and promoted a normal gut microbiome profile by partially attenuating the impacts of HFD.Bacteroidales S24-7,Alistipes,Lactobacillus, andRuminococcaceaeabundances were altered, andBacteroidales S24-7andAlistipesabundances were higher in the HFD+NaNO3group than that in the HFD group.

Conclusions

Inorganic dietary nitrate alleviated HFD-induced obesity and ameliorated disrupted glucolipid metabolism via NO3--NO2--NO pathway activation and gut microbiome modulation.
Association between race/ethnicity and the risk of amputation of lower extremities among medicare beneficiaries with diabetic foot ulcers and diabetic foot infections
Authors: Tan Tze-Woei , Armstrong David G , Concha-Moore Kirsten C , Marrero David G , Zhou Wei , Calhoun Elizabeth , Chang Ching-Yuan , Lo-Ciganic Wei-Hsuan
Published: August 2020

Introduction

This study aimed to examine the association of race and ethnicity on the risk of lower extremity amputations among Medicare beneficiaries with diabetic foot ulcers (DFUs) and diabetic foot infections (DFIs).

Research design and methods

A retrospective study included 2011–2015 data of a 5% sample of fee-for-service Medicare beneficiaries with a newly diagnosed DFU and/or DFI. The primary outcome was the time to the first major amputation episode after a DFU and/or DFI were identified using the diagnosis and procedure codes. We used multivariable Cox proportional hazards models to estimate the risk of time to the first major amputation across races, adjusting for sociodemographic and health status factors. Adjusted hazard ratios (aHRs) with a 95% CI were reported.

Results

Among 92 929 Medicare beneficiaries newly diagnosed with DFUs and/or DFIs, 77% were whites, 14.3% African Americans (AAs), 3.3% Hispanics, 0.7% Native Americans (NAs), and 4.0% were other races. The incidence rates of major amputation were 0.02 person-years for NAs, 0.02 person-years for AAs, 0.01 person-years for Hispanics, 0.01 person-years for other races, and 0.01 person-years for whites (p<0.05). Multivariable analysis showed that AAs (aHR=1.9, 95% CI 1.7 to 2.2, p<0.0001) and NAs (aHR=1.8, 95% CI 1.3 to 2.6, p=0.001) were associated with an increased risk of major amputation compared with whites. Beneficiaries with DFUs and/or DFIs diagnosed by a podiatrist or primary care physician (aHR=0.7, 95% CI 0.6 to 0.8, p<0.0001, specialists as reference) or at an outpatient visit (aHR=0.3, 95% CI 0.3 to 0.3, p<0.0001, inpatient stay as reference) were associated with a decreased risk of major amputation.

Conclusions

Racial and ethnic disparities in the risk of lower extremity amputations appear to exist among fee-for-service Medicare beneficiaries with diabetic foot problems. AAs and NAs with DFUs and/or DFIs were associated with an increased risk of major amputations compared with white Medicare beneficiaries.
Ultrasound-guided bilateral pudendal nerve blocks of nulliparous women with epidural labour analgesia in the second stage of labour: a randomised, double-blind, controlled trial
Authors: Xu Jialing , Zhou Riyong , Su Weijue , Wang Shi , Xia Yun , Papadimos Thomas , Zhao Junzhao , Xu Xuzhong
Published: August 2020

Objective

To explore whether an ultrasound-guided pudendal nerve block (PNB) could decrease anaesthetic use, thereby shortening the length of the second stage of labour in women undergoing epidural analgesia.

Design

Prospective, single-centre, randomised, double-blind, controlled trial.

Setting

An obstetric centre in a general hospital in China.

Participants

72 nulliparous women were randomised, and 71 women completed the study.

Intervention

An ultrasound-guided bilateral PNB was administered to all study participants; the PNB group were given 0.25% ropivacaine 10 mL, while the control group were given 10 mL saline.

Main outcome measure

The primary outcome measure was the duration of the second stage of labour. Secondary outcomes included additional bolus administration, total hourly bupivacaine consumption, difference in thickness between the contracted and relaxed rectus abdominis muscle before (DRAM1) and 30 min after (DRAM2) PNB, urge to defecate, maternal cooperation, preservation of the lower limb motor function, tightness of the perineum, and Numeric Rating Scale (NRS) score for pain.

Results

The duration of the second stage of labour was shorter in the PNB group than in the control group (difference of 33.8 min (95% CI 15.6 to 52.0), p<0.001). Additional bolus administration and total hourly bupivacaine consumption were lower in the PNB group than in the control group (p<0.001). DRAM2 was greater (p<0.001), rate of parturient women with the urge to defecate was higher (p=0.014), maternal cooperation was superior (p=0.002), and lower limb motor function preservation was greater (p=0.004) in the PNB group relative to the control group. Tightness of the perineum was eliminated from the results due to the inconsistent application of the criteria by the nursing staff. There was no significant difference in NRS scores between the groups.

Conclusions

Nulliparous women with epidural analgesia who received an ultrasound-guided bilateral PNB may reduce their need for bupivacaine and consequently shorten the length of the second stage of labour, therein indicating that a bilateral PNB may serve as an additional effective adjunct method of labour analgesia.

Trial registration number

ChiCTR-IOR-16009121.
Prevalence of and rationale for antimicrobial prescription during ambulatory care visits in Japan: a prospective, multicentre, cross-sectional study
Authors: Komagamine Junpei , Kobayashi Masaki , Mori Takahiro
Published: August 2020

Objective

To determine the rate of outpatient antimicrobial use and the rationale for antimicrobial prescription.

Design

A prospective, multicentre, cross-sectional study.

Setting

Ambulatory care settings at community general hospitals.

Participants

A total of 1972 consecutive ambulatory visits by 1952 patients were included from 2 February 2020 to 13 February 2020. Visits resulting in hospital admission and regularly scheduled visits were excluded.

Main outcome measures

The primary outcome was the proportion of ambulatory visits resulting in antimicrobial drug prescriptions. The secondary outcomes were the reasons for antimicrobial drug prescription and the proportion of unnecessary antimicrobial prescriptions among all antimicrobial drugs used for treatment.

Results

The mean patient age was 53.8 (SD 25.8) years old, and the proportion of women was 52.6%. A total of 162 antimicrobial drugs were prescribed in 153 (7.8%) visits. The most common antimicrobial drugs were penicillins (n=48, 29.6%), followed by third-generation cephalosporins (n=35, 21.6%) and quinolones (n=20, 12.4%). Among all the antimicrobial drugs prescribed, 125 (77.2%), 18 (11.1%) and 11 (6.8%) were used for infection treatment, wound prophylaxis and surgical prophylaxis, respectively. Of the 125 antimicrobial drugs used for infection treatment, 60 (48.0%) were judged to be unnecessary.

Conclusions

One in every 13 ambulatory visits resulted in antimicrobial use in Japan. Three-fourths of the prescribed antimicrobial drugs were used for infection treatment, but approximately half of those drugs may have been unnecessary. Further efforts to reduce unnecessary antimicrobial drug use are needed.

Trial registration number

UMIN000039360.
Psychometric properties of the Urdu version of the Hospital Anxiety and Depression Scale (HADS) among pregnant women in Abbottabad, Pakistan
Authors: Lodhi Fahad Saqib , Elsous Aymen M , Irum Saadia , Khan Adeel Ahmed , Rabbani Unaib
Published: August 2020

Background

The Hospital Anxiety and Depression Scale (HADS) is a widely used instrument to measure anxiety and depression symptoms.

Aims

This study aimed to translate, validate and test the applicability of the Urdu version of the Hospital Anxiety and Depression Scale (HADS/UV) among pregnant women.

Methods

The original English version of the HADS was translated into Urdu by three bilingual experts and retranslated to English using the forward–backward approach. The questionnaire was administered to a sample of 200 pregnant women availing obstetrics and gynaecology services for routine prenatal check-ups of Ayub Teaching Hospital, Abbottabad, Pakistan. Psychometric properties of the instrument, including reliability (internal consistency, test–retest analysis and interitems correlation), were tested. Face and content validity were also assessed. Content Validity Index (CVI) was determined using the average approach and Item-Level Content Validity Index (I-CVI) and Scale-Level Content Validity Index (S-CVI) were calculated accordingly. Construct validity was examined through exploratory factor analysis.

Results

Cronbach’s alpha coefficient has been found to be 0.82 for the anxiety subscale and 0.64 for the depression subscale, while overall alpha of the HADS/UV is 0.84. The Urdu version is content valid, and the S-CVI of anxiety subscale, depression subscale and HADS/UV are 0.947, 948 and 0.947, respectively. Test–retest reliability is 0.884 and 0.934 as measured by Pearson correlation and intraclass correlation, respectively. HADS/UV items correlated positively with the whole scale (p<0.001). Factor analysis with varimax rotation revealed that two factors explained 42.75% of the variance. Items’ distribution was quite similar to the original HADS.

Conclusion

The HADS/UV is a psychometrically sound instrument with satisfactory measurement, including good internal consistency. The instrument shows promise to be a sound tool to assess anxiety and depression in pregnancy.
Impact of significant paravalvular leaks after transcatheter aortic valve implantation on anaemia and mortality
Authors: Lopez-Pais Javier , Lopez-Otero Diego , Garcia-Touchard Arturo , Izquierdo Coronel Barbara , Antúnez Muiños Pablo José , Cia Mendioroz Xabier , Pérez-Poza Marta , Otero Garcia Óscar , Martinez Peredo Carlos Gustavo , Arellano Flores Carlos , Álvarez-Rodríguez Leyre , Gomez Peña Fernando , Juskova Mária , Sanmartín Pena Juan Carlos , Estevez-Loureiro Rodrigo , Cid Álvarez Ana Belén , Trillo Nouche Ramiro , Alonso-Martín Joaquín Jesús , González-Juanatey Jose Ramón , Oteo Juan Francisco
Published: August 2020

Objective

The aim of this work is to assess the relationship between significant paravalvular leak (SPL) after transcatheter aortic valve implantation (TAVI) on anaemia and their impact on prognosis.

Methods

Observational analytic study developed at two university hospitals, including all consecutive patients who underwent TAVI during a 10-year period (2009 to 2018). A logistic regression model was created to determine independent predictors of anaemia at 3 months. Time to event outcomes were analysed with Cox regression. Median follow-up was 21.3±21.9 months.

Results

788 patients were included. 5.3% had SPL. SPL was an independent predictor of anaemia 3 months after TAVI (OR: 8.31, 95% CI: 2.06 to 33.50). SPL and anaemia at 3 months were independently associated with long-term mortality (HR: 1.82, 95% CI: 1.16 to 2.85; HR: 2.07, 95% CI: 1.39 to 3.08).

Conclusion

SPL is an independent predictor of anaemia at 3 months after TAVI, a condition that doubles long-term mortality. Our findings could explain in part the worse prognosis of SPL after TAVI. Further pathophysiological studies are necessary to explain this association.
Role of Microrna-7 in Liver Diseases: A Comprehensive Review of the Mechanisms and Therapeutic Applications
Authors: Han Sen , Zhang Ting , Kusumanchi Praveen , Huda Nazmul , Jiang Yanchao , Liangpunsakul Suthat , Yang Zhihong
Published: August 2020

MicroRNA-7 (miR-7) is a small non-coding RNA, which plays critical roles in regulating gene expression of multiple key cellular processes. MiR-7 exhibits a tissue-specific pattern of expression, with abundant levels found in the brain, spleen, and pancreas. Although it is expressed at lower levels in other tissues, including the liver, miR-7 is involved in both the development of organs and biological functions of cells. In this review, we focus on the mechanisms by which miR-7 controls cell growth, proliferation, invasion, metastasis, metabolism, and inflammation. We also summarize the specific roles of miR-7 in liver diseases. MiR-7 is considered as a tumor suppressor miRNA in hepatocellular carcinoma and is involved in the pathogenesis of hepatic steatosis and hepatitis. Future studies to further define miR-7 functions and its mechanism in association with other types of liver diseases should be explored. An improved understanding from these studies will provide us a useful perspective leading to mechanism-based intervention by targeting miR-7 for the treatment of liver diseases.
Preclinical investigations and a first-in-human phase I trial of M4112, the first dual inhibitor of indoleamine 2,3-dioxygenase 1 and tryptophan 2,3-dioxygenase 2, in patients with advanced solid tumors
Authors: Naing Aung , Eder Joseph P , Piha-Paul Sarina A , Gimmi Claude , Hussey Elizabeth , Zhang Sen , Hildebrand Vera , Hosagrahara Vinayak , Habermehl Christina , Moisan Jacques , Papadopoulos Kyriakos P
Published: August 2020

Background

M4112 is an oral, potent, and selective indoleamine 2,3-dioxygenase 1 (IDO1) and tryptophan 2,3-dioxygenase 2 (TDO2) dual inhibitor. Here, we report preclinical data and first-in-human phase I data, including safety, tolerability, pharmacokinetics, pharmacodynamics, and preliminary efficacy, of M4112 monotherapy in patients with advanced solid tumors.

Methods

In preclinical studies, M4112 was administered to mice with IDO1-expressing tumors to determine tumor IDO1 and liver TDO2 inhibition. In the phase I trial, patients received doses of M4112 two times per day in 28-day cycles until progression, toxicity, or withdrawal of consent. The primary objective was to determine the maximum tolerated dose (MTD) and recommended phase II dose (RP2D). The primary endpoint was the incidence of dose-limiting toxicities (DLTs), treatment-emergent adverse events (TEAEs), and treatment-emergent changes in safety parameters. Other endpoints included pharmacokinetics, pharmacodynamics, and antitumor effects.

Results

In mice, M4112 significantly decreased the kynurenine:tryptophan ratio in the liver and tumor. Fifteen patients received M4112 at five distinct dose levels (three patients per cohort: 100, 200, 400, 600, and 800 mg two times per day orally). Initially, all doses inhibited IDO1 ex vivo, but plasma kynurenine levels returned to or exceeded baseline levels after day 15. Despite initial changes in kynurenine, there was no significant reduction of plasma kynurenine at steady state. There was one DLT (grade 3 allergic dermatitis; 800 mg two times per day) and one grade 2 QT prolongation (800 mg two times per day), resulting in dose reduction (not a DLT). M4112 was well tolerated, and neither the MTD nor the RP2D was established. TEAEs included fatigue, nausea, and vomiting. The best overall response was stable disease (n=9, 60%).

Conclusions

There were no serious safety concerns at any dose. Although M4112 inhibited IDO1 activity ex vivo, plasma kynurenine levels were not reduced despite achieving target exposure.

Trial registration numberNCT03306420.
The Great Debate at ‘Immunotherapy Bridge’, Naples, December 5, 2019
Authors: Ascierto Paolo A , Bifulco Carlo , Galon Jerome , Garbe Claus , Khleif Samir N , McQuade Jennifer , Odunsi Kunle , Okada Hideho , Paulos Chrystal M , Quezada Sergio A , Tawbi Hussein A , Timmerman John , Trinchieri Giorgio , Butterfield Lisa H , Puzanov Igor
Published: August 2020

As part of the 2019 Immunotherapy Bridge congress (December 4–5, Naples, Italy), the Great Debate session featured counterpoint views from leading experts on six topical issues in immunotherapy today. These were the use of chimeric antigen receptor T cell therapy in solid tumors, whether the Immunoscore should be more widely used in clinical practice, whether antibody-dependent cellular cytotoxicity is important in the mode of action of anticytotoxic T-lymphocyte-associated protein 4 antibodies, whether the brain is immunologically unique or just another organ, the role of microbiome versus nutrition in affecting responses to immunotherapy, and whether chemotherapy is immunostimulatory or immunosuppressive. Discussion of these important topics are summarized in this report.
Assessing performance of pathogenicity predictors using clinically relevant variant datasets
Authors: Gunning Adam C , Fryer Verity , Fasham James , Crosby Andrew H , Ellard Sian , Baple Emma L , Wright Caroline F
Published: August 2020

Background

Pathogenicity predictors are integral to genomic variant interpretation but, despite their widespread usage, an independent validation of performance using a clinically relevant dataset has not been undertaken.

Methods

We derive two validation datasets: an ‘open’ dataset containing variants extracted from publicly available databases, similar to those commonly applied in previous benchmarking exercises, and a ‘clinically representative’ dataset containing variants identified through research/diagnostic exome and panel sequencing. Using these datasets, we evaluate the performance of three recent meta-predictors, REVEL, GAVIN and ClinPred, and compare their performance against two commonly used in silico tools, SIFT and PolyPhen-2.

Results

Although the newer meta-predictors outperform the older tools, the performance of all pathogenicity predictors is substantially lower in the clinically representative dataset. Using our clinically relevant dataset, REVEL performed best with an area under the receiver operating characteristic curve of 0.82. Using a concordance-based approach based on a consensus of multiple tools reduces the performance due to both discordance between tools and false concordance where tools make common misclassification. Analysis of tool feature usage may give an insight into the tool performance and misclassification.

Conclusion

Our results support the adoption of meta-predictors over traditional in silico tools, but do not support a consensus-based approach as in current practice.
Psychological impact of COVID-19 pandemic on postgraduate trainees: a cross-sectional survey
Authors: Imran Nazish , Masood Hafiz Muhammad Umar , Ayub Maryam , Gondal Khalid Masood
Published: August 2020

Background

The present study aimed to evaluate psychological impact of COVID-19 outbreak on postgraduate trainees in Pakistan by quantifying the symptoms of depression, anxiety and acute stress disorder and by analysing potential risk factors associated with these symptoms.

Methods

Following Institutional Review Board approval, a cross-sectional study was conducted among 10,178 postgraduate trainees following COVID-19 outbreak through e-log system of College of Physicians and Surgeons of Pakistan. The nine-item Patient Health Questionnaire, seven-item Generalised Anxiety Disorder scale and Stanford Acute Stress Reaction Questionnaire were used to collect data. Statistical analyses were conducted using SPSS.26. Descriptive statistics, Mann-Whitney U test, the χ2 test and logistic regression analysis were performed. The significance level was set at α=0.05.

Results

The prevalence of depressive symptoms, generalised anxiety disorder and acute stress disorder were 26.4%, 22.6% and 4.4%, respectively. Female postgraduate trainees, senior trainees and front-line workers reported experiencing more anxiety, depression and acute stress symptoms (p value<0.001). Logistic regression showed that being a front-line and senior staff member and female was associated with higher risk of experiencing symptoms of depression, anxiety and acute stress.

Conclusions

Our study findings raise concerns about the psychological well-being of postgraduate trainees during the acute COVID-19 outbreak in Pakistan. It is necessary to employ strategies to minimise the psychological distress and provide adequate psychosocial support for postgraduate trainees during crisis situation such as COVID-19 pandemic.
Impact of deprivation and comorbidity on outcomes in emergency general surgery: an epidemiological study
Authors: Wohlgemut Jared M , Ramsay George , Griffin Russell L , Jansen Jan O
Published: August 2020

Background

The impact of socioeconomic deprivation and comorbidities on the outcome of patients who require emergency general surgery (EGS) admission is poorly understood. The aim of this study was to examine the effect of deprivation and comorbidity on mortality, discharge destination and length of hospital stay (LOS) in patients undergoing EGS in Scotland.

Methods

Prospectively collected data from all Scottish adult patients (aged >15 years) requiring EGS admitted between 1997 and 2016 were obtained from the Scottish Government. Data included age, sex, Scottish Index of Multiple Deprivation (SIMD), 5-year Charlson Comorbidity Index (CCI), whether an operation took place and outcomes including mortality, discharge destination and LOS. Logistic regression was used for the analysis of mortality and discharge destination and Poisson regression was used for LOS.

Results

1 477 810 EGS admissions were analyzed. 16.2% were in the most deprived SIMD decile and 5.6% in the least deprived SIMD decile. 75.6% had no comorbidity, 20.3% had mild comorbidity, 2.5% had moderate comorbidity and 1.6% had severe comorbidity. 78.6% were discharged directly home. Inpatient, 30-day, 90-day and 1-year crude mortality was 1.7%, 3.7%, 7.2% and 12.4%, respectively. Logistic regression showed that severe comorbidity was associated with not being discharged directly to home (OR 0.38, 95% CI 0.37 to 0.39) and higher inpatient mortality (OR 13.74, 95% CI 13.09 to 14.42). Compared with the most affluent population, the most deprived population were less likely to be discharged directly to home (OR 0.97, 95% CI 0.95 to 0.99) and had higher inpatient mortality (OR 1.36, 95% CI 1.8 to 1.46). Poisson analysis showed that severe comorbidity (OR 1.69, 95% CI 1.68 to 1.69) and socioeconomic deprivation (OR 1.11, 95% CI 1.11 to 1.12) were associated with longer LOS.

Discussion

Increased levels of comorbidity and, to a lesser extent, socioeconomic deprivation are key drivers of mortality, discharge destination and LOS following admission to an EGS service.

Level of evidence

III (prospective/retrospective with up to two negative criteria).

Study type

Epidemiological/prognostic.
Unusual severe fungal pneumonia in Vietnamese potbelly pigs: two cases
Authors: Heller Meera , Busch Roselle , Koehne Amanda , Keating M Kelly , Mohr Charles
Published: August 2020

Two 18‐month‐old, castrated male Vientamese potbelly pigs presented for diagnosis and treatment of respiratory disease of two to three weeks duration which had been non‐responsive to antibiotic therapy. The pigs were unrelated, from separate households and presented roughly a year apart. Both pigs presented in the summer months, and were from the mid‐central valley of California. Diagnostics included complete blood count, serum biochemistry, thoracic radiographs, thoracic ultrasound examination and ultrasound‐guided thoracocentesis. Findings included severe pleural effusion and large thoracic masses in both cases. Thoracocentesis revealed suppurative exudate in one case and mixed inflammatory exudate in the second; both cases were negative on bacterial culture. Treatment in each case was limited as the severity of the disease lead to euthanasia of the patients. Definitive diagnosis of granulomatous fungal pneumonia was obtained on postmortem examination of both patients.
Systemic and mucosal levels of lactoferrin in very low birth weight infants supplemented with bovine lactoferrin
Authors: Itell Hannah L. , Berenz Andrew , Mangan Riley J. , Permar Sallie R. , Kaufman David A.
Published: August 2020

Lactoferrin supplementation may help prevent infections in preterm infants, but the efficacy has varied with different doses and products. We assessed the absorption and excretion of bovine lactoferrin (bLF) in 31 infants receiving 100, 200, or 300 mg·kg–1·day–1 of enteral bLF for 30 days. bLF and human lactoferrin (hLF) in infant saliva, blood, urine, and stool, as well as expressed (EBM) or donor breast milk (DBM) that were collected (i) before the treatment was initiated, (ii) at study day 22, and (iii) one week after treatment cessation, were measured using ELISA. During treatment, bLF was absorbed from the gastrointestinal tract and detected in plasma, saliva, and urine, as well as excreted in stool. Levels of bLF in the saliva and stool began to decline within 12 h after dosing, and bLF was undetectable in all samples one week after treatment. The concentrations of hLF exceeded those of bLF across sample types and time-points. Infants receiving EBM demonstrated higher levels of hLF in the saliva and stool than those receiving DBM. Neither bLF nor hLF levels varied by patient characteristics, bLF dosage, or infection status. This is the first study demonstrating bLF absorption into the bloodstream and distribution to saliva and urine in preterm infants. Future studies should further explore LF pharmacokinetics because higher and more frequent dosing may improve the clinical benefit of LF supplementation.
Effects of Saccharomyces cerevisiae fermentation products and subacute ruminal acidosis on feed intake, fermentation, and nutrient digestibilities in lactating dairy cows
Authors: Khalouei H. , Seranatne V. , Fehr K. , Guo J. , Yoon I. , Khafipour E. , Plaizier J.C.
Published: August 2020

Effects of Saccharomyces cerevisiae fermentation products (SCFP) and subacute ruminal acidosis (SARA) on rumen and hindgut fermentation, feed intake, and total tract nutrient digestibilities were determined in 32 lactating Holstein cows between weeks 4 and 9 of lactation. Treatments included control, 14 g·d−1 Diamond V Original XPC™ (SCFPa; Diamond V, Cedar Rapids, IA, USA), 19 g·d−1 NutriTek® (SCFPb-1X; Diamond V), and 38 g·d−1 NutriTek® (SCFPb-2X; Diamond V). During weeks 5 and 8, SARA challenges were conducted by switching from a 18.6% to a 27.9% dry matter (DM) starch diet. This reduced the rumen and feces pH. The durations of the rumen pH below 5.6 during these challenges averaged 175.0, 233.8, 246.9, and 79.3 min·d−1 for the control, SCFPa, SCFPb-1X, and SCFPb-2X treatments, respectively. Hence, SARA was not induced under the SCFPb-2X treatment. The feces pH during the SARA challenges was lowest during SCFPb-2X, suggesting this treatment shifted fermentation from the rumen to the hindgut. The SARA challenges reduced the total tract digestibility of DM, neutral detergent fiber digestibility (NDFd), and phosphorus, but tended to increase that of starch. The SCFPb-2X treatment increased the NDFd from 52.7% to 61.8% (P < 0.05). The SCFPb-2X treatment attenuated impacts of SARA.