bluuebunny/crossref_metadata_2025 huggingface dataset:
Drawing and storing blood using a 3D printed bottle cap and a disinfected 500 mL drinking bottle: A proof‐of‐concept study
Authors: Rognhaug Martin , Eliassen Håkon Skogrand
Published: April 2024

Background

Today, with wars raging in Ukraine and the Middle East, the demand for blood is high. Despite this, few companies produce the necessary equipment to draw, store, and transfuse whole blood. This study evaluated the safety and performance of a 3D printed bottle cap in conjunction with a water bottle and some available consumables to draw and store fresh whole blood.

Study Design and Methods

Bags of saline, and freshly donated whole blood, was transferred to the water bottle through a 3D printed bottle cap and stored for 72 h. An identical setup, transferring saline to a Terumo blood collection bag was used as control. Performance and safety were evaluated by calculating infusion rate and observing for backflow, respectively. The blood was also tested for hemolysis and bacterial growth at four sampling points.

Results

The cap‐and‐bottle setup was faster than control in terms of flow rate when transferring saline (1.53 vs. 1.81 mL/s, p < .001), and non‐inferior to saline control when transfusing blood (1.53 vs. 1.49 mL/s, p = .641). We did not observe any risks of causing the donor iatrogenic harm, and there was no evidence of increased hemolysis. However, there were traces of bacterial contamination in three of six bottles.

Conclusion

This study indicates that drawing blood is both feasible and safe, utilizing a 3D printed cap and bottle setup. Flow rate was faster than control, and mechanical properties of the blood were not affected. We were unable to determine the source of bacterial contamination in the blood.
Can nematode infection cause internal bleeding in dogs? A case of Dirofilaria immitis infection in cavitary fluids
Authors: Alonso Flavio H. , Shapiro Karen , Camp Lauren E.
Published: April 2024

A 9‐year‐old dog was presented with weight loss, respiratory effort, and an enlarged abdomen. Imaging studies and exploratory surgery showed pulmonary and splenic masses and bi‐cavitary effusion, later classified as hemorrhage. Cytology of the peritoneal and pleural fluids also revealed several microfilariae. Immunologic and molecular analyses confirmed Dirofilaria immitis infection and histopathology of the spleen indicated a cavernous endothelial proliferation with undefined etiology (hemangiosarcoma vs reaction to parasite infestation). The nematode larvae are speculated to have entered body cavities via erratic migration or via hemorrhage and visceral lesions to be related to parasitism. Nematode infection should be considered as a differential diagnosis for internal bleeding of undetermined origin.
Evaluation of mediastinoscopy for cranial mediastinal and tracheobronchial lymphadenectomy in canine cadavers
Authors: Gibson Erin A. , Brust Kelsey , Steffey Michele A.
Published: April 2024

Objective

To report technical feasibility and describe procedural details of a novel single incision minimally invasive approach to the mediastinum in cadaver dogs.

Study design

Cadaveric study.

Animals

Large breed (25–40 kg) cadaver dogs (n = 10).

Methods

Three of 10 cadavers were used for preliminary technique development without data recording. Cadaver specimens underwent pre‐ and postoperative thoracic computed tomographic scans. Seven dogs were placed in dorsal recumbency and mediastinoscopy was performed via a SILS port placed cranial to the thoracic inlet with CO2 insufflation of the mediastinum at 2–4 mmHg. Retrieval of all CT and visually identified mediastinal lymph nodes (LN) was attempted; endoscopic compartmental and individual LN dissection times and subjective operative challenges were recorded. Procedural success scores for visualization and dissection as well as NASA‐task force index scores were recorded per lymph node, per cadaver.

Results

Median time required for initial approach including SILS placement was 5 min (range 5–10 min). Individual LN retrieval times ranged from 2 to 32 min. Mediastinoscopic retrieval of LNs was most commonly successful for the left tracheobronchial LN (7/7), followed by the right tracheobronchial LN (4/7), the left and right sternal LNs (3/7 each), and the cranial mediastinal LNs (1/7). Post‐procedure pleural gas was identified on CT in 4/7 cadavers.

Conclusions

Mediastinoscopy as reported was feasible in large breed canine cadavers and retrieval or cup biopsy of a variety of lymph nodes is possible from the described approach. Application in living animals and its associated challenges should be further investigated.

Clinical significance

Mediastinoscopy may provide a novel minimally invasive approach to the evaluation and oncologic staging of the cranial mediastinum in dogs.
Combined action observation and mental imagery versus neuromuscular electrical stimulation as novel therapeutics during short‐term knee immobilization
Authors: Harmon Kylie K. , Girts Ryan M. , Rodriguez Gabriela , Beausejour Jonathan P. , Pagan Jason I. , Carr Joshua C. , Garcia Jeanette , Roberts Michael D. , Hahs‐Vaughn Debbie L. , Stout Jeffrey R. , Fukuda David H. , Stock Matt S.
Published: April 2024

Limb immobilization causes rapid declines in muscle strength and mass. Given the role of the nervous system in immobilization‐induced weakness, targeted interventions may be able to preserve muscle strength, but not mass, and vice versa. The purpose of this study was to assess the effects of two distinct interventions during 1 week of knee joint immobilization on muscle strength (isometric and concentric isokinetic peak torque), mass (bioimpedance spectroscopy and ultrasonography), and neuromuscular function (transcranial magnetic stimulation and interpolated twitch technique). Thirty‐nine healthy, college‐aged adults (21 males, 18 females) were randomized into one of four groups: immobilization only (n = 9), immobilization + action observation/mental imagery (AOMI) (n = 10), immobilization + neuromuscular electrical stimulation (NMES) (n = 12), or control group (n = 8). The AOMI group performed daily video observation and mental imagery of knee extensions. The NMES group performed twice daily stimulation of the quadriceps femoris. Based on observed effect sizes, it appears that AOMI shows promise as a means of preserving voluntary strength, which may be modulated by neural adaptations. Strength increased from PRE to POST in the AOMI group, with +7.2% (Cohen's d = 1.018) increase in concentric isokinetic peak torque at 30°/s. However, NMES did not preserve muscle mass. Though preliminary, our findings highlight the specific nature of clinical interventions and suggest that muscle strength can be independently targeted during rehabilitation. This study was prospectively registered: ClinicalTrials.gov NCT05072652.
Specificity of early motor unit adaptations with resistive exercise training
Authors: Del Vecchio Alessandro , Enoka Roger Maro , Farina Dario
Published: April 2024

After exposure of the human body to resistive exercise, the force‐generation capacity of the trained muscles increases significantly. Despite decades of research, the neural and muscular stimuli that initiate these changes in muscle force are not yet fully understood. The study of these adaptations is further complicated by the fact that the changes may be partly specific to the training task. For example, short‐term strength training does not always influence the neural drive to muscles during the early phase (<100 ms) of force development in rapid isometric contractions. Here we discuss some of the studies that have investigated neuromuscular adaptations underlying changes in maximal force and rate of force development produced by different strength training interventions, with a focus on changes observed at the level of spinal motor neurons. We discuss the different motor unit adjustments needed to increase force or speed, and the specificity of some of the adaptations elicited by differences in the training tasks.

image
Lactate and hydrogen ions play a predominant role in evoking the exercise pressor reflex during ischaemic contractions but not during freely perfused contractions
Authors: Ducrocq Guillaume P. , Anselmi Laura , Ruiz‐Velasco Victor , Kaufman Marc P.
Published: April 2024

We investigated the role played by lactate and hydrogen in evoking the exercise pressor reflex (EPR) in decerebrated rats whose hindlimb muscles were either freely perfused or ischaemic. Production of lactate and hydrogen by the contracting hindlimb muscles was manipulated by knocking out the myophosphorylase gene (pygm). In knockout rats (pygm−/−; n = 13) or wild‐type rats (pygm+/+; n = 13), the EPR was evoked by isometrically contracting the triceps surae muscles. Blood pressure, tension, blood flow, renal sympathetic nerve activity and blood lactate concentrations were measured. Intramuscular metabolites and pH changes induced by the contractions were quantified by 31P‐magnetic resonance spectroscopy (n = 5). In a subset of pygm−/− rats (n = 5), contractions were evoked with prior infusion of lactate (pH 6.0) in an attempt to restore the effect of lactate and hydrogen ions. Contraction of freely perfused muscles increased blood lactate and decreased muscle pH in pygm+/+ rats only. Despite these differences, the reflex pressor and sympathetic responses to freely perfused contraction did not differ between groups (P = 0.992). During ischaemia, contraction increased muscle lactate and hydrogen ion production in pygm+/+ rats (P < 0.0134), whereas it had no effect in pygm−/− rats (P > 0.783). Likewise, ischaemia exaggerated the reflex pressor, and sympathetic responses to contraction in pygm+/+ but not in pygm−/− rats. This exaggeration was restored when a solution of lactate (pH 6.0) was infused prior to the contraction in pygm−/− rats. We conclude that lactate and hydrogen accumulation in contracting myocytes play a key role in evoking the metabolic component of the EPR during ischaemic but not during freely perfused contractions.

image

Key points

Conflicting results exist about the role played by lactate and hydrogen ions in evoking the exercise pressor reflex.

Using CRISP‐Cas9, we rendered the myophosphorylase gene non‐functional to block the production of lactate and hydrogen ions. The exercise pressor reflex was evoked in decerebrated rats by statically contracting the triceps surae muscles with or without muscle ischaemia.

Static contraction elevated the concentration of lactate and hydrogen ions in pygm+/+ but not in pygm−/− rats. Despite these differences, the exercise pressor reflex was not different between groups.

Acute muscle ischaemia exaggerated the concentration of lactate and hydrogen ions in pygm+/+ but not in pygm−/− rats. Likewise, acute muscle ischaemia exaggerated the exercise pressor reflex in pygm+/+ but not in pygm−/− rats.

We conclude that lactate and hydrogen play a key role in evoking the exercise pressor reflex during ischaemic but not during freely perfused contractions.
New species of oribatid mites (Acari, Oribatida) of Scheloribates (Perscheloribates) (Scheloribatidae) from Vietnam
Authors: Ermilov Sergey G.
Published: April 2024

Two new species of the subgenus Scheloribates (Perscheloribates) (Oribatida, Scheloribatidae) are described, based on adults collected from dry litter in Shorea dipterocarp forest in Southern Vietnam. Scheloribates (P.) posteriorugosus sp. nov. is characterized by the presence of heavily rugose sculpturing (several longitudinal ridges with deep furrows between them) in posterior part of the notogaster; S. (P.) dentiunguis sp. nov. is characterized by the presence of strong teeth in mediobasal part of the leg claws. An identification key to the known species of S. (Perscheloribates) from the Oriental region is presented.
Design and application of universal primers for mitochondrial genome sequencing of synanthropic mite species (Astigmatina: Acaridae)

Authors: Wu Jingjing , Zhuo Mo , Hu Caixiao , Pan Ruoxi , Ding Lan , Wu Xuechun , Ye Changjiang , Tao Xianglin , Sun Entao , Liu Ying
Published: April 2024

This study aims to design and develop a set of universal primers for amplification of the entire mitochondrial genome of Acaridae mites based on polymerase chain reaction (PCR). The conserved regions were screened by comparing the mitochondrial genomes of known Acaridae mites in GenBank. To ensure that the primers cover the entire mitochondrial sequence, the mitochondrial genome was divided into six overlapping fragments based on the conserved region. After synthesizing primers, we extracted the total DNA of Acaridae mites collected from grain stores. Then the total DNA of individual Acaridae mites was used as the template for PCR, Sanger sequencing, and sequence splicing. Agarose gel electrophoresis revealed that the lengths of the six amplification products were between 1.2 kb and 3.8 kb, consistent with design expectations. The resulting sequences could be successfully spliced to form a loop structure, confirming that the primers cover the entire mitochondrial genome. The splicing results were consistent with the full sequence released in GenBank. Developed universal primers can achieve rapid amplification and help to obtain mitochondrial genome from a single acarid mite with high efficiency.
African American English speakers’ pitch variation and rate adjustments for imagined technological and human addressees
Authors: Cohn Michelle , Mengesha Zion , Lahav Michal , Heldreth Courtney
Published: April 2024

This paper examines the adaptations African American English speakers make when imagining talking to a voice assistant, compared to a close friend/family member and to a stranger. Results show that speakers slowed their rate and produced less pitch variation in voice-assistant-“directed speech” (DS), relative to human-DS. These adjustments were not mediated by how often participants reported experiencing errors with automatic speech recognition. Overall, this paper addresses a limitation in the types of language varieties explored when examining technology-DS registers and contributes to our understanding of the dynamics of human-computer interaction.
Effect of Mindful Sport Performance Enhancement in College Athletes for Reducing Sports-Caused Anxiety and Improving Self-Awareness: A Critically Appraised Topic
Authors: Garg Shivam , Uriegas Nancy A. , Winkelmann Zachary K. , Adams Morgan , Fraley Amy L.
Published: April 2024

Mindful Sport Performance Enhancement (MSPE) training is a relatively new concept, which focuses on helping athletes manage a variety of stressors experienced throughout a season, including performing well academically, staying fit, having a productive season in their sport, and maintaining a healthy social life. A need for a critical appraisal is needed to assess the effectiveness of the intervention. Two cohort studies and one randomized control trial were included in the study and assessed using STROBE and PEDro Scale. Key results show, all 3 studies identified participants experiencing benefits after MSPE with aspects of awareness, acceptance, and emotion regulation. Furthermore, student-athletes who attended either all the sessions or more sessions after the 6-week course showed greater satisfaction with mental and physical health. Overall, there is level “B” evidence to support effectiveness of MSPE for college athletes in reducing sport anxiety and improving their overall well-being.
Targeting the postsynaptic scaffolding protein PSD-95 enhances BDNF signaling to mitigate depression-like behaviors in mice
Authors: Shi Xin , Zhou Xiao-zhong , Chen Gang , Luo Wei-feng , Zhou Chengyu , He Tian-ju , Naik Mandar T. , Jiang Qin , Marshall John , Cao Cong
Published: April 2024

Signaling mediated by brain-derived neurotrophic factor (BDNF), which is supported by the postsynaptic scaffolding protein PSD-95, has antidepressant effects. Conversely, clinical depression is associated with reduced BDNF signaling. We found that peptidomimetic compounds that bind to PSD-95 promoted signaling by the BDNF receptor TrkB in the hippocampus and reduced depression-like behaviors in mice. The compounds CN2097 and Syn3 both bind to the PDZ3 domain of PSD-95, and Syn3 also binds to an α-helical region of the protein. Syn3 reduced depression-like behaviors in two mouse models of stress-induced depression; CN2097 had similar but less potent effects. In hippocampal neurons, application of Syn3 enhanced the formation of TrkB–Gα
i1/3
–PSD-95 complexes and potentiated downstream PI3K-Akt-mTOR signaling. In mice subjected to chronic mild stress (CMS), systemic administration of Syn3 reversed the CMS-induced, depression-associated changes in PI3K-Akt-mTOR signaling, dendrite complexity, spine density, and autophagy in the hippocampus and reduced depression-like behaviors. Knocking out Gα
i1/3
in hippocampal neurons prevented the therapeutic effects of Syn3, indicating dependence of these effects on the TrkB pathway. The findings suggest that compounds that induce the formation of PSD-95–TrkB complexes have therapeutic potential to alleviate depression.
Nude blebs expose cells to phagocytes
Authors: VanHook Annalisa M.
Published: April 2024

Displacement of the glycocalyx by membrane blebbing enables macrophages to recognize apoptotic cells.
Doxycycline, levofloxacin, and moxifloxacin are superior to ciprofloxacin in treating anthrax meningitis in rabbits and NHP
Authors: Ben-Shmuel Amir , Glinert Itai , Sittner Assa , Bar-David Elad , Schlomovitz Josef , Levy Haim , Weiss Shay
Published: April 2024

Efficient treatment of anthrax-related meningitis in patients poses a significant therapeutic challenge. Previously, we demonstrated in our anthrax meningitis rabbit model that ciprofloxacin treatment is ineffective with most of the treated animals succumbing to the infection. Herein we tested the efficacy of doxycycline in our rabbit model and found it highly effective. Since all of our findings are based on a rabbit model, we test the efficacy of ciprofloxacin or doxycycline in a specific central nervous system (CNS) model developed in non-human primates (NHPs). Similar to rabbits, ciprofloxacin treatment was ineffective, while doxycycline protected the infected rhesus macaques (
n
= 2) from the lethal CNS
Bacillus anthracis
infection. To test whether the low efficacy of Ciprofloxacin is an example of low efficacy of all fluoroquinolones or only this substance, we treated rabbits that were inoculated intracisterna magna (ICM) with levofloxacin or moxifloxacin. We found that in contrast to ciprofloxacin, levofloxacin and moxifloxacin were highly efficacious in treating lethal anthrax-related meningitis in rabbits and NHP (levofloxacin). We demonstrated (in naïve rabbits) that this difference probably results from variances in blood–brain-barrier penetration of the different fluoroquinolones. The combined treatment of doxycycline and any one of the tested fluoroquinolones was highly effective in the rabbit CNS infection model. The combined treatment of doxycycline and levofloxacin was effective in an inhalation rabbit model, as good as the doxycycline mono-therapy. These findings imply that while ciprofloxacin is highly effective as a post-exposure prophylactic drug, using this drug to treat symptomatic patients should be reconsidered.
The interplay between electron transport chain function and iron regulatory factors influences melanin formation in
Cryptococcus neoformans
Authors: Xue Peng , Sánchez-León Eddy , Hu Guanggan , Lee Christopher W. J. , Black Braydon , Brisland Anna , Li Haohua , Jung Won Hee , Kronstad James W.
Published: April 2024

Mitochondrial functions are critical for the ability of the fungal pathogen
Cryptococcus neoformans
to cause disease. However, mechanistic connections between key functions such as the mitochondrial electron transport chain (ETC) and virulence factor elaboration have yet to be thoroughly characterized. Here, we observed that inhibition of ETC complex III suppressed melanin formation, a major virulence factor. This inhibition was partially overcome by defects in Cir1 or HapX, two transcription factors that regulate iron acquisition and use. In this regard, loss of Cir1 derepresses the expression of laccase genes as a potential mechanism to restore melanin, while HapX may condition melanin formation by controlling oxidative stress. We hypothesize that ETC dysfunction alters redox homeostasis to influence melanin formation. Consistent with this idea, inhibition of growth by hydrogen peroxide was exacerbated in the presence of the melanin substrate L-DOPA. In addition, loss of the mitochondrial chaperone Mrj1, which influences the activity of ETC complex III and reduces ROS accumulation, also partially overcame antimycin A inhibition of melanin. The phenotypic impact of mitochondrial dysfunction was consistent with RNA-Seq analyses of WT cells treated with antimycin A or L-DOPA, or cells lacking Cir1 that revealed influences on transcripts encoding mitochondrial functions (e.g., ETC components and proteins for Fe-S cluster assembly). Overall, these findings reveal mitochondria-nuclear communication
via
ROS and iron regulators to control virulence factor production in
C. neoformans
.

IMPORTANCE

There is a growing appreciation of the importance of mitochondrial functions and iron homeostasis in the ability of fungal pathogens to sense the vertebrate host environment and cause disease. Many mitochondrial functions such as heme and iron-sulfur cluster biosynthesis, and the electron transport chain (ETC), are dependent on iron. Connections between factors that regulate iron homeostasis and mitochondrial activities are known in model yeasts and are emerging for fungal pathogens. In this study, we identified connections between iron regulatory transcription factors (e.g., Cir1 and HapX) and the activity of complex III of the ETC that influence the formation of melanin, a key virulence factor in the pathogenic fungus
Cryptococcus neoformans
. This fungus causes meningoencephalitis in immunocompromised people and is a major threat to the HIV/AIDS population. Thus, understanding how mitochondrial functions influence virulence may support new therapeutic approaches to combat diseases caused by
C. neoformans
and other fungi.
UV-C side-emitting optical fiber-based disinfection: a promising approach for infection control in tight channels
Authors: Mohsin Muhammad Salman , Avdic Melisa , Fitzpatrick Katrina , Lanzarini-Lopes Mariana
Published: April 2024

The growth of pathogenic bacteria in moist and wet surfaces and tubing of medically relevant devices results in serious infections in immunocompromised patients. In this study, we investigated and demonstrated the successful implementation of a UV-C side-emitting optical fiber in disinfecting medically relevant pathogenic bacteria (
Pseudomonas aeruginosa
and Methicillin-resistant
Staphylococcus aureus
[MRSA]) within tight channels of polytetrafluoroethylene (PTFE). PTFE is a commonly used material both in point-of-use (POU) water treatment technologies and medical devices (dental unit water line [DUWL], endoscope). For a 1-m-long PTFE channel, up to ≥6 log inactivation was achieved using a 1-m-long UV side-emitting optical fiber (SEOF) with continuous 16-h exposure of low UV-C radiation ranging from ~0.23 to ~29.30 μW/cm
2
. Furthermore, a linear model was used to calculate the inhibition zone constant (k`), which enables us to establish a correlation between UV dosage and the extent of inactivated surface area (cm
2
) for surface-bound
Escherichia coli
on a nutrient-rich medium. The k` value for an irradiance ranging from ~150 to ~271.50 μW/cm
2
was calculated to be 0.564 ± 0.6 cm·cm
2
/mJ. This study demonstrated the efficacy of SEOFs for disinfection of medically relevant microorganisms present in medically and domestically relevant tight channels. The impact of the results in this study extends to the optimization of operational efficiency in pre-existing UV surface disinfection setups that currently operate at UV dosages exceeding the optimal levels.

IMPORTANCE

Germicidal UV radiation has gained global recognition for its effectiveness in water and surface disinfection. Recently, various works have illustrated the benefit of using UV-C side-emitting optical fibers (SEOFs) for the disinfection of tight polytetrafluoroethylene (PTFE) channels. This study now demonstrates its impact for disinfection of medically relevant organisms and introduces critical design calculations needed for its implementation. The flexible geometry and controlled emission of light in these UV-SEOFs make them ideal for light distribution in tight channels. Moreover, the results presented in this manuscript provide a novel framework that can be employed in various applications, addressing microbial contamination and the disinfection of tight channels.
SARS-CoV-2 cellular and humoral responses in vaccine-naive individuals during the first two waves of COVID-19 infections in the southern region of The Netherlands: a cross-sectional population-based study
Authors: Hanssen D. A. T. , Arts K. , Nix W. H. V. , Sweelssen N. N. B. , Welbers T. T. J. , de Theije C. , Wieten L. , Pagen D. M. E. , Brinkhues S. , Penders J. , Dukers-Muijrers N. H. T. M. , Hoebe C. J. P. A. , Savelkoul P. H. M. , van Loo I. H. M.
Published: April 2024

With the emergence of highly transmissible variants of concern, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) still poses a global threat of coronavirus disease 2019 (COVID-19) resurgence. Cellular responses to novel variants are more robustly maintained than humoral responses, and therefore, cellular responses are of interest in assessing immune protection against severe disease in the population. We aimed to assess cellular responses to SARS-CoV-2 at the population level. IFN
γ
(interferon
γ
) responses to wild-type SARS-CoV-2 were analyzed using an ELISpot assay in vaccine-naive individuals with different humoral responses: Ig (IgM and/or IgG) seronegative (
n
= 90) and seropositive (
n
= 181) with low (<300 U/mL) or high (≥300 U/mL) humoral responses to the spike receptor binding domain (anti-S-RBD). Among the seropositive participants, 71.3% (129/181) were IFN
γ
ELISpot positive, compared to 15.6% (14/90) among the seronegative participants. Common COVID-19 symptoms such as fever and ageusia were associated with IFN
γ
ELISpot positivity in seropositive participants, whereas no participant characteristics were associated with IFN
γ
ELISpot positivity in seronegative participants. Fever and/or dyspnea and anti-S-RBD levels were associated with higher IFN
γ
responses. Symptoms of more severe disease and higher anti-S-RBD responses were associated with higher IFN
γ
responses. A significant proportion (15.6%) of seronegative participants had a positive IFN
γ
ELISpot. Assessment of cellular responses may improve estimates of the immune response to SARS-CoV-2 in the general population.

IMPORTANCE

Data on adaptive cellular immunity are of interest to define immune protection against severe acute respiratory syndrome coronavirus 2 in a population, which is important for decision-making on booster-vaccination strategies. This study provides data on associations between participant characteristics and cellular immune responses in vaccine-naive individuals with different humoral responses.
Minimum inhibitory concentrations of azithromycin in clinical isolates of
Mycobacterium avium
complex in Japan
Authors: Uwamino Yoshifumi , Aoki Wataru , Inose Rika , Kamoshita Yuka , Mikita Kei , Namkoong Ho , Nishimura Tomoyasu , Matsushita Hiromichi , Hasegawa Naoki
Published: April 2024

The latest guidelines include azithromycin as a preferred regimen for treating
Mycobacterium avium
complex (MAC) pulmonary disease. However, serially collected susceptibility data on clinical MAC isolates are limited, and no breakpoints have been determined. We investigated the minimum inhibitory concentrations (MICs) of azithromycin and clarithromycin for all MAC strains isolated in 2021 from a single center in Japan, excluding duplicates. The MICs were determined using a panel based on the microbroth dilution method, according to the latest Clinical and Laboratory Standards Institute recommendations. The MICs were determined for 318 MAC strains. Although there was a significant positive correlation between the MICs of azithromycin and clarithromycin, the MICs of azithromycin tended to be higher than those of clarithromycin. Among the cases in which the strains were isolated, 18 patients initiated treatment, including azithromycin treatment, after sample collection. Some patients infected with stains with relatively high azithromycin MICs achieved a microbiological cure with azithromycin-containing regimens. This study revealed a higher MIC distribution for azithromycin than clarithromycin, raising questions about the current practice of estimating azithromycin susceptibility based on the clarithromycin susceptibility test result. However, this was a single-center study that included only a limited number of cases treated with azithromycin. Therefore, further multicenter studies that include a greater number of cases treated with azithromycin are warranted to verify the distribution of azithromycin MICs and examine the correlation between azithromycin MICs and treatment effectiveness.

IMPORTANCE

The macrolides serve as key drugs in the treatment of pulmonary
Mycobacterium avium
complex infection, and the administration of macrolide should be guided by susceptibility test results. Azithromycin is recommended as a preferred choice among macrolides, surpassing clarithromycin; however, drug susceptibility testing is often not conducted, and clarithromycin susceptibility is used as a surrogate. This study represents the first investigation into the minimum inhibitory concentration of azithromycin on a scale of several hundred clinical isolates, revealing an overall tendency for higher minimum inhibitory concentrations compared with clarithromycin. The results raise questions about the appropriateness of using clarithromycin susceptibility test outcomes for determining the administration of azithromycin. This study highlights the need for future discussions on the clinical breakpoints of azithromycin, based on large-scale clinical research correlating azithromycin susceptibility with treatment outcomes.
Mutation of the peptide-regulated transcription factor ComR for amidated peptide specificity and heterologous function in
Lactiplantibacillus plantarum
WCFS1
Authors: Brasino Michael , Wagnell Eli , Ozdemir E. Sila , Ranganathan Srivathsan , Merritt Justin
Published: April 2024

There is a growing interest in the use of probiotic bacteria as biosensors for the detection of disease. However, there is a lack of bacterial receptors developed for specific disease biomarkers. Here, we have investigated the use of the peptide-regulated transcription factor ComR from S
treptococcus spp
. for specific peptide biomarker detection. ComR exhibits a number of attractive features that are potentially exploitable to create a biomolecular switch for engineered biosensor circuitry within the probiotic organism
Lactiplantibacillus plantarum
WCFS1. Through iterative design-build-test cycles, we developed a genomically integrated, ComR-based biosensor circuit that allowed WCFS1 to detect low nanomolar concentrations of ComR’s cognate peptide XIP. By screening a library of ComR proteins with mutant residues substituted at the K100 position
,
we identified mutations that increased the specificity of ComR toward an amidated version of its cognate peptide, demonstrating the potential for ComR to detect this important class of biomarker.

IMPORTANCE

Using bacteria to detect disease is an exciting possibility under active study. Detecting extracellular peptides with specific amino acid sequences would be particularly useful as these are important markers of health and disease (biomarkers). In this work, we show that a probiotic bacteria (
Lactiplantibacillus plantarum
) can be genetically engineered to detect specific extracellular peptides using the protein ComR from Streptococcus bacteria. In its natural form, ComR allowed the probiotic bacteria to detect a specific peptide, XIP. We then modified XIP to be more like the peptide biomarkers found in humans and engineered ComR so that it activated with this modified XIP and not the original XIP. This newly engineered ComR also worked in the probiotic bacteria, as expected. This suggests that with additional engineering, ComR might be able to activate with human peptide biomarkers and be used by genetically engineered probiotic bacteria to better detect disease.
Early-life gut microbiota associates with allergic rhinitis during 13-year follow-up in a Finnish probiotic intervention cohort
Authors: Kallio Sampo , Jian Ching , Korpela Katri , Kukkonen Anna Kaarina , Salonen Anne , Savilahti Erkki , Kuitunen Mikael , M. de Vos Willem
Published: April 2024

Perinatal and early-life factors reported to affect risk of allergic diseases may be mediated by changes in the gut microbiota. Here, we explored the associations between the infant gut microbiota and allergic morbidity in childhood until 13 years of age in a subgroup of the FLORA probiotic intervention cohort. A mixture of four probiotic strains with galacto-oligosaccharides was administrated to the mothers from the 36th week of the pregnancy and later to their infants until 6 months of age. The infants were monitored for the manifestations of atopic eczema, food allergy, allergic rhinitis, and asthma by a pediatrician at 2 and 5 years of age; the allergic status was subsequently verified by a questionnaire at 10 and 13 years of age. The fecal microbiota at 3 months was profiled by 16S rRNA amplicon sequencing targeting the V3-V4 region, with and without adjusting for potentially important early-life factors. Overall, the positive diagnosis for allergic rhinitis between 2 and 13 years was associated with microbiota composition both in non-adjusted and adjusted models. This association was more pronounced in children born to one parent with confirmed atopic diseases compared to those who had two atopic parents and was characterized by a lower relative abundance of
Bifidobacterium
and
Escherichia/Shigella spp
. and a higher proportion of
Bacteroides
. While the probiotic and galacto-oligosaccharides intervention in the entire cohort was previously shown to reduce the prevalence of eczema to a certain extent, no associations were found between the 3-month gut microbiota and childhood eczema in the studied sub-cohort.

IMPORTANCE

Allergic diseases have increased in prevalence during the past decades globally. Although probiotics have been considered a promising strategy for preventing certain allergy related symptoms, studies connecting the infant gut microbiota and later life allergic morbidity in various populations remain limited. The present study supports an association between the infant microbiota and allergic morbidity after first years of life, which has been rarely examined.

CLINICAL TRIALS

Registered at ClinicalTrials.gov
(NCT00298337)
.
Genome sequence-based species classification of
Enterobacter cloacae
complex: a study among clinical isolates
Authors: Qiu Xuemei , Ye Kun , Ma Yanning , Zhao Qiang , Wang Lifeng , Yang Jiyong
Published: April 2024

Accurate species-level identification of
Enterobacter cloacae
complex (ECC) is crucial for related research. The classification of ECC is based on strain-to-strain phylogenetic congruence, as well as genomic features including average nucleotide identity (ANI) and digitalized DNA-DNA hybridization (dDDH). ANI and dDDH derived from whole-genome sequencing have emerged as a reliable metric for assessing genetic relatedness between genomes and are increasingly recognized as a standard for species delimitation. Up to now, there are two different classification methods for ECC. The first one categorizes
E. hormaechei
, a species within ECC, into five subspecies (
E. hormaechei
subsp.
steigerwaltii,
subsp.
oharae,
subsp.
xiangfangensis,
subsp.
hoffmannii,
and subsp.
hormaechei
). The second classifies
E. hormaechei
as three species:
E. hormaechei, “E. xiangfangensis,” “E. hoffmanii.”
While the former is well-accepted in the academic area, the latter may have a greater ability to distinguish different species of ECC. To assess the suitability of these identification criteria for clinical ECC isolates, we conducted a comprehensive analysis involving phylogenetic analysis, ANI and dDDH value alignment, virulence gene identification, and capsule typing on 256 clinical ECC strains isolated from the bloodstream. Our findings indicated that the method of categorizing
E. hormaechei
into five subspecies has better correlation and consistency with the molecular characteristics of clinical ECC isolates, as evidenced by phylogenetic analysis, virulence genes, and capsule typing. Therefore, the subspecies-based classification method appears more suitable for taxonomic assignments of clinical ECC isolates.

IMPORTANCE

Standardizing taxonomy of the
Enterobacter cloacae
complex (ECC) is necessary for data integration across diverse studies. The study utilized whole-genome data to accurately identify 256 clinical ECC isolated from bloodstream infections using average nucleotide identity (ANI), digitalized DNA-DNA hybridization (dDDH), and phylogenetic analysis. Through comprehensive assessments including phylogenetic analysis, ANI and dDDH comparisons, virulence gene, and capsule typing of the 256 clinical isolates, it was concluded that the classification method based on subspecies exhibited better correlation and consistency with the molecular characteristics of clinical ECC isolates. In summary, this research contributes to the precise identification of clinical ECC at the species level and expands our understanding of ECC.
Risk of cancer in patients with bile acid diarrhoea: a Danish nationwide matched cohort study
Authors: Nyboe Andersen Nynne , Wildt Signe , Iversen Aske Thorn , Poulsen Gry , Jess Tine , Munck Lars Kristian , Borup Christian
Published: April 2024

Objective

Bile acid diarrhoea is a common cause of chronic diarrhoea. Increased levels of potentially carcinogenic bile acids in faeces, theoretically, may increase the risk of colorectal cancer in particular, but the long-term disease course is unknown. We aimed to investigate the overall and site-specific cancer risk in bile acid diarrhoea.

Design

Adult patients with bile acid diarrhoea were identified using nationwide Danish registries from 2003 to 2020 by a diagnostic gold-standard 75-selenium tauroselcholic acid procedure followed within 6 months by sequestrant prescription. The risk of overall and site-specific cancers in cases with bile acid diarrhoea was compared with sex, age and comorbidity-adjusted matched controls. A competing risk model estimated cumulative incidence functions and cause-specific HRs.

Results

We identified 2260 patients with bile acid diarrhoea with a mean follow-up of 5.5 years (SD 4.2). The overall cancer risk was increased by an HR of 1.32 (95% CI 1.12 to 1.54). The risk of site-specific cancer was increased in 3 of 10 cancer groups: haematological, HR 2.41 (1.36 to 4.02); skin, HR 1.33 (1.01 to 1.71); and male genital cancers, HR 1.85 (1.11 to 2.92). No increased risk of colorectal cancer was detected in patients with bile acid diarrhoea, HR 0.73 (0.34 to 1.63).

Conclusions

Bile acid diarrhoea was associated with an increased overall risk of cancer, especially haematological cancers, but the risk of colorectal cancer was not increased. The lack of a diagnostic code for bile acid diarrhoea and potential residual confounding are limitations, and the findings should be replicated in other cohorts.
Real-world efficacy, roll-out and uptake of intramuscular tixagevimab/cilgavimab as COVID-19 pre-exposure prophylaxis in people with multiple sclerosis and neuroimmunological conditions during the COVID-19 pandemic
Authors: Rath Louise , Yeh Wei Zhen , Roldan Angie , Wesselingh Robb , Zhong Michael , Tan Tracie , Seery Nabil , Bridge Francesca , Foong YiChao , Skibina Olga , Nesbitt Cassie , Butzkueven Helmut , Monif Mastura , van der Walt Anneke
Published: April 2024

Background

In Australia, tixagevimab/cilgavimab 150 mg/150 mg was a government-funded pre-exposure prophylaxis for COVID-19 people with multiple sclerosis (pwMS) and other neuroimmunological conditions (pwNIc) treated with anti-CD20 antibodies or sphingosine-1-phosphate receptor modulators were eligible.

Objective

To analyse the roll-out, uptake and real-world efficacy of tixagevimab/cilgavimab in the prevention and severity of COVID-19. To assess compliance with uptake depending on the location of delivery.

Methods

We undertook a single-centre study. 440 pwMS and pwNIc were eligible. Logistic regression was used to assess predictors of COVID-19 during follow-up and to assess predictors of uptake among those who consented.

Results

Of the eligible pwMS and pwNIc in our service, 52.7% (233/440) requested a consultation and were included in this study. Consultation resulted in 71.7% of people (167/233) receiving the treatment. Of these, 94.0% (157/167) had received three or more COVID-19 vaccines. Among those who received a single dose of tixagevimab/cilgavimab, 19.16% (32/167) tested positive for COVID-19 during the observational window. The majority of these were on ocrelizumab (68.8% (22/32)). None of those with COVID-19 required hospitalisation or supplemental oxygen. There was no difference in odds of COVID-19 during the observation period between those who received and did not receive tixagevimab/cilgavimab (adjusted OR, aOR 2.16 (95% CI 0.82 to 6.85), p=0.43). Uptake of tixagevimab/cilgavimab was highest when offered at the hospital infusion centre (aOR 3.09 (95% CI 1.08 to 9.94) relative to referral to the local pharmacy, p=0.04).

Conclusion

Tixagevimab/cilgavimab administration did not protect against subsequent COVID-19 in our cohort. Compliance with uptake was influenced by administration location.
Repurposing of dextromethorphan as an adjunct therapy in patients with major depressive disorder: a randomised, group sequential adaptive design, controlled clinical trial protocol
Authors: Maji Shampa , Mohapatra Debadatta , Jena Monalisa , Srinivasan Anand , Maiti Rituparna
Published: April 2024

Background

Therapeutic latency, lack of efficacy and adverse drug reactions are the major concerns in current antidepressant therapies. To overcome these treatment hurdles, add-on therapy to conventional antidepressant medications may lead to better therapeutic outcomes. The present randomised controlled trial has been planned to evaluate the efficacy and safety of add-on dextromethorphan to selective serotonin reuptake inhibitors (SSRIs) in major depressive disorder (MDD).

Methods and analysis

A randomised, double-blind, add-on, placebo-controlled, group sequential design clinical trial will be conducted on patients with MDD who will be randomly assigned to the control and the test group in a 1:1 ratio. Patients in the test group will get dextromethorphan 30 mg once daily, whereas patients in the control group will receive a placebo once daily as an add-on to ongoing SSRI treatment for 8 weeks. All patients will be evaluated for the primary outcome (change in the Montgomery-Åsberg Depression Rating Scale score) and secondary outcomes (treatment response rate, remission rate, Clinical Global Impression, serum brain-derived neurotrophic factor, serum dextromethorphan and treatment-emergent adverse events) over the period of 8 weeks. Intention-to-treat analysis will be done for all parameters using suitable statistical tools.

Ethics and dissemination

This study was approved by the Institutional Ethics Committee of All India Institute of Medical Sciences, Bhubaneswar, India, and the study conformed to the provisions of the Declaration of Helsinki and ICMR’s ethical guidelines for biomedical research on human subjects (2017). Written informed consent will be obtained from the participants before recruitment. The results of this study will be published in peer-reviewed publications.

Trial registration number

NCT05181527.
Social inequality in prevalence of NCD risk factors: a cross-sectional analysis from the population-based Tromsø Study 2015–2016
Authors: Hetland Rebecca A , Wilsgaard Tom , Hopstock Laila Arnesdatter , Ariansen Inger , Johansson Jonas , Jacobsen Bjarne K , Grimsgaard Sameline
Published: April 2024

Objective

We aimed to examine associations between educational level, serving as an indicator of socioeconomic position, and prevalence of WHO-established leading behavioural and biological risk factors for non-communicable diseases (NCDs), in middle-aged to older women and men.

Design

Population-based cross-sectional study.

Setting

All inhabitants of the municipality of Tromsø, Norway, aged ≥40 years, were invited to the seventh survey (2015–2016) of the Tromsø Study; an ongoing population-based cohort study.

Participants

Of the 32 591 invited; 65% attended, and a total of 21 069 women (53%) and men aged 40–99 years were included in our study.

Outcome measures

We assessed associations between educational level and NCD behavioural and biological risk factors: daily smoking, physical inactivity (sedentary in leisure time), insufficient fruit/vegetable intake (<5 units/day), harmful alcohol use (>10 g/day in women, >20 g/day in men), hypertension, obesity, intermediate hyperglycaemia and hypercholesterolaemia. These were expressed as odds ratios (OR) per unit decrease in educational level, with 95% CIs, in women and men.

Results

In women (results were not significantly different in men), we observed statistically significant associations between lower educational levels and higher odds of daily smoking (OR 1.69; 95% CI 1.60 to 1.78), physical inactivity (OR 1.38; 95% CI 1.31 to 1.46), insufficient fruit/vegetable intake (OR 1.54, 95% CI 1.43 to 1.66), hypertension (OR 1.25; 95% CI 1.20 to 1.30), obesity (OR 1.23; 95% CI 1.18 to 1.29), intermediate hyperglycaemia (OR 1.12; 95% CI 1.06 to 1.19), and hypercholesterolaemia (OR 1.07; 95% CI 1.03 to 1.12), and lower odds of harmful alcohol use (OR 0.75; 95% CI 0.72 to 0.78).

Conclusion

We found statistically significant educational gradients in women and men for all WHO-established leading NCD risk factors within a Nordic middle-aged to older general population. The prevalence of all risk factors increased at lower educational levels, except for harmful alcohol use, which increased at higher educational levels.