Hydroponic Growth of [13C]-Labeled Tobacco for DNA Damage Studies in Cigarette Smokers.

Cigarette smoking is the acknowledged major cause of cancers of the lung and oral cavity and is an established important risk factor for multiple other cancers. DNA addition products (DNA adducts) caused by cigarette smoking are critical factors in its mechanism of carcinogenesis. However, most DNA adducts detected to date in humans cannot be specifically ascribed to smoking but rather have multiple exogenous and endogenous sources. In the study reported here, we prepared [13C]-labeled tobacco to address this problem. We report for the first time the successful growth from seeds to flowering under hydroponic conditions of highly [13C]-labeled tobacco in a controlled 13CO2 environment. The standard growth procedure with optimized conditions is described in detail. The [13C]-enrichment rate was assessed by quantifying nicotine and sugars and their [13C]-isotopologues in this tobacco using high-resolution mass spectrometry, reaching >94% in the tobacco leaves. The [13C]-labeled leaves after curing will be used to make cigarettes, allowing investigation of the specific contributions of tobacco smoke carcinogens to identified DNA adducts in smokers.

Diversity of structure and function in Cullin E3 ligases.

The cellular process by which the protein ubiquitin (Ub) is covalently attached to a protein substrate involves Ub activating (E1s) and conjugating enzymes (E2s) that work together with a large variety of E3 ligases that impart substrate specificity. The largest family of E3s is the Cullin-RING ligase (CRL) family which utilizes a wide variety of substrate receptors, adapter proteins, and cooperating ligases. Cryo-electron microscopy (cryoEM) has revealed a wide variety of structures which suggest how Ub transfer occurs. Hydrogen deuterium exchange mass spectrometry (HDXMS) has revealed the role of dynamics and expanded our knowledge of how covalent NEDD8 modification (neddylation) activates the CRLs, particularly by facilitating cooperation with additional RING-between-RING ligases to transfer Ub.

Inhibition of Acyl-CoA Synthetase Long Chain Isozymes Decreases Multiple Myeloma Cell Proliferation and Causes Mitochondrial Dysfunction.

Multiple myeloma (MM) is an incurable cancer of plasma cells with a 5-year survival rate of 59%. Dysregulation of fatty acid (FA) metabolism is associated with MM development and progression; however, the underlying mechanisms remain unclear. Acyl-CoA synthetase long-chain family members (ACSLs) convert free long-chain fatty acids into fatty acyl-CoA esters and play key roles in catabolic and anabolic fatty acid metabolism. The Cancer Dependency Map data suggested that ACSL3 and ACSL4 were among the top 25% Hallmark Fatty Acid Metabolism genes that support MM fitness. Here, we show that inhibition of ACSLs in human myeloma cell lines using the pharmacological inhibitor Triascin C (TriC) causes apoptosis and decreases proliferation in a dose- and time-dependent manner. RNA-seq of MM.1S cells treated with TriC for 24 h showed a significant enrichment in apoptosis, ferroptosis, and ER stress. Proteomics of MM.1S cells treated with TriC for 48 h revealed that mitochondrial dysfunction and oxidative phosphorylation were significantly enriched pathways of interest, consistent with our observations of decreased mitochondrial membrane potential and increased mitochondrial superoxide levels. Interestingly, MM.1S cells treated with TriC for 24 h also showed decreased mitochondrial ATP production rates and overall lower cellular respiration.

Study protocol for a comparative randomized controlled trial of Tai Chi and conventional exercise training on alleviating depression in older insomniacs.

Background: Insomnia and depression are prevalent mental disorders that are often comorbid among older adults. Lifestyle intervention strategies incorporating Tai Chi or conventional exercise have been shown to alleviate symptoms of insomnia and depression. However, the comparative efficacy of these exercise modalities in individuals with both disorders has yet to be determined. Therefore, the aim of this study is to examine the efficacy of Tai Chi and conventional exercise for reducing depressive symptoms in older adults with chronic insomnia and depressive symptoms, when compared to a health education control. Methods: This study is a prospective, assessor-blinded, three-arm, parallel group, randomized controlled trial. Older adults aged ≥60 years with a diagnosis of chronic insomnia and depressive symptoms will be randomly assigned to a Tai Chi, conventional exercise or health education control condition on a 1:1:1 basis. Interventions will last for 3 months, with a 6-month follow-up period. The primary outcome is depressive symptoms, assessed using the Hospital Anxiety and Depression Scale. Secondary outcomes include subjective sleep quality, 7-day actigraphy, 7-day sleep diary, anxiety symptoms, quality of life, medication usage and physical function. All measurements will be conducted at baseline, 3 months and 9 months by outcome assessors who are blinded to group allocation. Discussion: This study will compare the efficacy of Tai Chi and conventional exercise in improving depression outcomes in older adults with chronic insomnia and depressive symptoms. Our results will shed light on the clinical potential of these interventions for combating insomnia and depression in older adults.

Protease-Induced Excitation of Dorsal Root Ganglion Neurons in Response to Acute Perturbation of the Gut Microbiota Is Associated With Visceral and Somatic Hypersensitivity.

BACKGROUND & AIMS: Abdominal pain is a major symptom of diseases that are associated with microbial dysbiosis, including irritable bowel syndrome and inflammatory bowel disease. Germ-free mice are more prone to abdominal pain than conventionally housed mice, and reconstitution of the microbiota in germ-free mice reduces abdominal pain sensitivity. However, the mechanisms underlying microbial modulation of pain remain elusive. We hypothesized that disruption of the intestinal microbiota modulates the excitability of peripheral nociceptive neurons. METHODS: In vivo and in vitro assays of visceral sensation were performed on mice treated with the nonabsorbable antibiotic vancomycin (50 µg/mL in drinking water) for 7 days and water-treated control mice. Bacterial dysbiosis was verified by 16s rRNA analysis of stool microbial composition. RESULTS: Treatment of mice with vancomycin led to an increased sensitivity to colonic distension in vivo and in vitro and hyperexcitability of dorsal root ganglion (DRG) neurons in vitro, compared with controls. Interestingly, hyperexcitability of DRG neurons was not restricted to those that innervated the gut, suggesting a widespread effect of gut dysbiosis on peripheral pain circuits. Consistent with this, mice treated with vancomycin were more sensitive than control mice to thermal stimuli applied to hind paws. Incubation of DRG neurons from naive mice in serum from vancomycin-treated mice increased DRG neuron excitability, suggesting that microbial dysbiosis alters circulating mediators that influence nociception. The cysteine protease inhibitor E64 (30 nmol/L) and the protease-activated receptor 2 (PAR-2) antagonist GB-83 (10 µmol/L) each blocked the increase in DRG neuron excitability in response to serum from vancomycin-treated mice, as did the knockout of PAR-2 in NaV1.8-expressing neurons. Stool supernatant, but not colonic supernatant, from mice treated with vancomycin increased DRG neuron excitability via cysteine protease activation of PAR-2. CONCLUSIONS: Together, these data suggest that gut microbial dysbiosis alters pain sensitivity and identify cysteine proteases as a potential mediator of this effect.

Evaluation of 5 Polymerase Chain Reaction Assays for the Detection of Mpox Virus.

BACKGROUND: In 2022, the global dissemination of mpox virus (MPXV) outside endemic regions prompted the expansion of diagnostic testing worldwide. This study assesses the performance characteristics of 5 real-time polymerase chain reaction (PCR) assays in detecting MPXV during the 2022 outbreak. METHODS: Clinical specimens collected from patients across Ontario, Canada, were tested on the following assays: RealStar Orthopoxyvirus PCR and FlexStar Monkeypox virus PCR (Altona Diagnostics), Novaplex MPXV (Seegene), VIASURE Monkeypox virus Real Time PCR Reagents (CerTest Biotec), and a laboratory-developed test. Positive percent agreement (PPA), negative percent agreement (NPA), relative limit of detection (LOD), and precision were evaluated and MPXV lineages were determined using an amplicon-based whole-genome sequencing (WGS) assay. RESULTS: Swabs were collected from various anatomic sites (65 positive and 30 negative). All assays demonstrated 100% NPA (95% confidence interval, 88.4%/88.1%-100.0%), with PPA ranging from 92.2% (82.7%-97.4%) to 96.9% (89.3%-99.6%). LOD and precision were comparable across assays, with coefficient of variations <3%. WGS analysis identified 6 lineages, all belonging to subclade IIb. CONCLUSIONS: The assays exhibited excellent PPA, NPA, LOD, and precision. Ongoing performance monitoring is essential to detect assay escape mutants and ensure universal detection of evolving MPXV strains.

Purification of functional mouse skeletal muscle mitochondria using percoll density gradient centrifugation.

OBJECTIVE: Our goal was to isolate purified mitochondria from mouse skeletal muscle using a Percoll density gradient and to assess bioenergetic function and purity via Seahorse Extracellular Flux (XF) Analyses and mass spectrometry. RESULTS: Mitochondria isolated from murine quadriceps femoris skeletal muscle using a Percoll density gradient method allowed for minimally contaminated preparations with time from tissue harvest to mitochondrial isolation and quantification in about 3-4 h. Percoll purification from 100 to 200 mg fresh tissue yielded ~ 200-400 ug protein. Mitochondrial bioenergetics evaluated using the Seahorse XFe96 analyzer, a high-throughput respirometry platform, showed optimum mitochondrial input at 500 ng with respiratory control ratio ranging from 3.9 to 7.1 using various substrates demonstrating a high degree of functionality. Furthermore, proteomic analysis of Percoll-enriched mitochondria isolated from skeletal muscle using this method showed significant enrichment of mitochondrial proteins indicating high sample purity. This study established a methodology that ensures sufficient high quality mitochondria for downstream analyses such as mitochondrial bioenergetics and proteomics.

Prevalence of Low-Frequency, Antiviral Resistance Variants in SARS-CoV-2 Isolates in Ontario, Canada, 2020-2023.

Importance: Nirmatrelvir-ritonavir is an oral antiviral medication that improves outcomes in SARS-CoV-2 infections. However, there is concern that antiviral resistance will develop and that these viruses could be selected for after treatment. Objective: To determine the prevalence of low-frequency SARS-CoV-2 variants in patient samples that could be selected for by nirmatrelvir-ritonavir. Design, Setting, and Participants: This retrospective cohort study was conducted at 4 laboratories that serve community hospitals, academic tertiary care centers, and COVID-19 assessment centers in Ontario, Canada. Participants included symptomatic or asymptomatic patients who tested positive for SARS-CoV-2 virus and submitted virus samples for diagnostic testing between March 2020 and January 2023. Exposure: SARS-CoV-2 infection. Main Outcomes and Measures: Samples with sufficient viral load underwent next-generation genome sequencing to identify low-frequency antiviral resistance variants that could not be identified through conventional sequencing. Results: This study included 78 866 clinical samples with next-generation whole-genome sequencing data for SARS-CoV-2. Low-frequency variants in the viral nsp5 gene were identified in 128 isolates (0.16%), and no single variant associated with antiviral resistance was predominate. Conclusions and Relevance: This cohort study of low-frequency variants resistant to nirmatrelvir-ritonavir found that these variants were very rare in samples from patients with SARS-CoV-2, suggesting that selection of these variants by nirmatrelvir-ritonavir following the initiation of treatment may also be rare. Surveillance efforts that involve sequencing of viral isolates should continue to monitor for novel resistance variants as nirmatrelvir-ritonavir is used more broadly.

Purification of functional mouse skeletal muscle mitochondria using Percoll density gradient centrifugation.

Objective: Our goal was to isolate purified mitochondria from mouse skeletal muscle using a Percoll density gradient and to assess bioenergetic function and purity via Seahorse Extracellular Flux (XF) Analyses and mass spectrometry. Results: Mitochondria isolated from murine quadriceps femoris skeletal muscle using a Percoll density gradient method allowed for minimally contaminated preparations with time from tissue harvest to mitochondrial isolation and quantification in about 3-4 hours. Percoll purification from 100-200 mg fresh tissue yielded ∼200-400 ug protein. Mitochondrial bioenergetics evaluated using the Seahorse XFe96 analyzer, a high-throughput respirometry platform, showed optimum mitochondrial input at 500 ng with respiratory control ratio ranging from 3.9-7.1 using various substrates demonstrating a high degree of functionality. Furthermore, proteomic analysis of Percoll-enriched mitochondria isolated from skeletal muscle using this method showed significant enrichment of mitochondrial proteins indicating high sample purity. This study established a methodology that ensures sufficient high quality mitochondria for downstream analyses such as mitochondrial bioenergetics and proteomics.

Towards a Vocal and Acoustic Description of Kapa Haka.

OBJECTIVE: The purpose of this study is to analyze the voice used in kapa haka, a contemporary indigenous vocal performance from New Zealand, which includes the well-known haka. This is the first study of its kind and is a preliminary investigation into the vocal and acoustic description of kapa haka. A significant goal of this study is to contribute ideas and potential definitions of vocal qualities to the community of kapa haka trainers that were specific to the genre. This strengths-based project raises up these vocal practices as legitimate and authentic colors within a vocal tradition that has seen generational learning disrupted by colonial interventions and is now flourishing within the community. METHODS: Eight kapa haka performers (three females, five males) were involved in the study, they were all experienced performers; furthermore, two had formal classical voice training. They were individually recorded speaking and performing three different genres of kapa haka (moteatea, waiata, and haka); all recordings were in te reo Maori (the Maori language). In addition, electroglottograph (EGG) signals were collected. An auditory-perceptual evaluation of the kapa haka voice was completed by three singer-researcher-pedagogues familiar with Western and non-western genres of singing. They all have experience appropriately collecting and analyzing data from indigenous communities, and they all understand the sociopolitical context of the vocal genre within the local colonial history. A specific evaluation instrument was created, and the results were validated. The acoustic and time-aligned EGG data was annotated at the phoneme level, and the signal analysis was performed in MATLAB. Averaged EGG pulses from /a/ segments were investigated, along with long-term average spectrums of the performances obtained from both the audio signal and the EGG signals. RESULTS: The perceptual analysis suggests the biggest difference in vocal styles was between the haka and the other two genres (and speech). The acoustic and EGG results support these findings. CONCLUSIONS: Common characteristics, perceptually and acoustically, were identified in the kapa haka performance styles across the eight performers.

Decreased CRISPLD2 expression impairs osteogenic differentiation of human mesenchymal stem cells during in vitro expansion.

Human mesenchymal stem cells (hMSCs) are the cornerstone of regenerative medicine; large quantities of hMSCs are required via in vitro expansion to meet therapeutic purposes. However, hMSCs quickly lose their osteogenic differentiation potential during in vitro expansion, which is a major roadblock to their clinical applications. In this study, we found that the osteogenic differentiation potential of human bone marrow stem cells (hBMSCs), dental pulp stem cells (hDPSCs), and adipose stem cells (hASCs) was severely impaired after in vitro expansion. To clarify the molecular mechanism underlying this in vitro expansion-related loss of osteogenic capacity in hMSCs, the transcriptome changes following in vitro expansion of these hMSCs were compared. Cysteine-rich secretory protein LCCL domain-containing 2 (CRISPLD2) was identified as the most downregulated gene shared by late passage hBMSCs, hDPSCs, and hASCs. Both the secreted and non-secreted CRISPLD2 proteins progressively declined in hMSCs during in vitro expansion when the cells gradually lost their osteogenic potential. We thus hypothesized that the expression of CRISPLD2 is critical for hMSCs to maintain their osteogenic differentiation potential during in vitro expansion. Our studies showed that the knockdown of CRISPLD2 in early passage hBMSCs inhibited the cells' osteogenic differentiation in a siRNA dose-dependent manner. Transcriptome analysis and immunoblotting indicated that the CRISPLD2 knockdown-induced osteogenesis suppression might be attributed to the downregulation of matrix metallopeptidase 1 (MMP1) and forkhead box Q1 (FOXQ1). Furthermore, adeno-associated virus (AAV)-mediated CRISPLD2 overexpression could somewhat rescue the impaired osteogenic differentiation of hBMSCs during in vitro expansion. These results revealed that the downregulation of CRISPLD2 contributes to the impaired osteogenic differentiation of hMSCs during in vitro expansion. Our findings shed light on understanding the loss of osteogenic differentiation in hMSCs and provide a potential therapeutic target gene for bone-related diseases.

Visual Aids for Sunscreen Application: A mixed methods study.

BACKGROUND: Consumers routinely underapply sunscreen, and this may be because they misinterpret application quantity directions on sunscreen labels. U.S. labels use ambiguous terms such as "apply generously" to communicate application quantity. Visual aids may be helpful to consumers to determine proper application quantity. PURPOSE: To examine sunscreen application practices and sunscreen quantity visual aid preferences. METHODS: Sets of five GIFs and five static images were created corresponding to five quantities of sunscreen. Participants extracted their typical-use sunscreen quantity from a pre-measured tube and answered questions about their estimation process and use of labeled instructions. Participants then identified their typical-use sunscreen quantity in response to three stimuli: teaspoons, images, and GIFs and reported their preferred format. RESULTS: No participants reported using the label directions for application quantity to inform their sunscreen use. The GIF-based visual aid for sunscreen application quantity (44%) was preferred over image-based (40%) or teaspoon-based (16%) versions (p = .0249). Addition of a GIF demonstration to clarify application quantity was reported as highly helpful. CONCLUSION: Consumers have difficulty determining the appropriate quantity of sunscreen to use and labeling does not influence their application quantity. A QR code incorporating an image or GIF demonstration of proper sunscreen quantity could be useful to sunscreen consumers.

Comparison of moderate and vigorous walking exercise on reducing depression in middle-aged and older adults: A pilot randomized controlled trial.

The optimal intensity of physical activity for alleviating depression in middle-aged and older adults remains unclear. The World Health Organization (WHO) physical activity guidelines recommend adults and older adults to accumulate at least 150-300 minutes of moderate or 75-150 minutes of vigorous aerobic-type physical activity weekly or an equivalent combination of both for health benefits including reduced risk of depression. This parallel, assessor-blinded, pilot randomized controlled trial preliminarily compared the effectiveness of the minimal volume of aerobic-type physical activity at different intensities as recommended by WHO (150 minutes of moderate walking exercise and 75 minutes of vigorous walking exercise weekly) on alleviating depression in middle-aged and older adults. Thirty-five participants were randomized to the control group (CON), moderate walking exercise group (MOD), or vigorous walking exercise group (VIG). The exercise frequency was three times a week and the intervention duration was 12 weeks. The primary outcome was the severity of depression assessed by Beck Depression Inventory. Secondary outcomes included severity of anxiety, sleep quality, quality of life, and cardiorespiratory fitness. Thirty participants completed the study (CON: n = 10, MOD: n = 10, VIG: n = 10). Participants in both MOD and VIG had significantly decreased depression severity after the intervention compared to CON (both p < 0.001). There was no significant difference between MOD and VIG (p = 0.92). Both MOD and VIG interventions also mitigated anxiety severity, improved quality of life and cardiorespiratory fitness. The minimum volume of walking exercise at either moderate or vigorous intensity was found to alleviate depression in middle-aged and older adults.Trial registration: ClinicalTrials.gov identifier: NCT04403373.HighlightsThe 12-week 150-minute moderate walking exercise and 75-minute vigorous walking exercise (the minimal weekly volumes of aerobic-type physical activity recommended by WHO guidelines) similarly reduced the severity of depression in middle-aged and older adults.The 12-week walking exercise interventions significantly reduced anxiety severity concomitant with improved quality of life and cardiorespiratory fitness in middle-aged and older adults with depression.

Comparison of SARS-CoV-2 Viral Loads in the Nasal Mucosa of Patients Infected With BA.1, BA.2, or BA.5 Omicron Lineages.

Lower viral loads were observed in the upper respiratory tract of patients infected with BA.1, whereas patients infected with BA.2 and BA.5 had comparable viral loads to those seen with Alpha or Delta. This suggests that viral loads are likely not responsible for the increased transmission of the Omicron lineages.

Female adolescent singing voice characteristics: an exploratory study using LTAS and inverse filtering.

Background and Aim: To date, little research is available that objectively quantifies female adolescent singing-voice characteristics in light of the physiological and functional developments that occur from puberty to adulthood. This exploratory study sought to augment the pool of data available that offers objective voice analysis of female singers in late adolescence.Methods: Using long-term average spectra (LTAS) and inverse filtering techniques, dynamic range and voice-source characteristics were determined in a cohort of vocally healthy cis-gender female adolescent singers (17 to 19 years) from high-school choirs in Aotearoa New Zealand. Non-parametric statistics were used to determine associations and significant differences.Results: Wide intersubject variation was seen between dynamic range, spectral measures of harmonic organisation (formant cluster prominence, FCP), noise components in the spectrum (high-frequency energy ratio, HFER), and the normalised amplitude quotient (NAQ) suggesting great variability in ability to control phonatory mechanisms such as subglottal pressure (Psub), glottal configuration and adduction, and vocal tract shaping. A strong association between the HFER and NAQ suggest that these non-invasive measures may offer complimentary insights into vocal function, specifically with regard to glottal adduction and turbulent noise in the voice signal.Conclusion: Knowledge of the range of variation within healthy adolescent singers is necessary for the development of effective and inclusive pedagogical practices, and for vocal-health professionals working with singers of this age. LTAS and inverse filtering are useful non-invasive tools for determining such characteristics.

Effects of exercise intensity and frequency on improving cognitive performance in middle-aged and older adults with mild cognitive impairment: A pilot randomized controlled trial on the minimum physical activity recommendation from WHO.

Background: The World Health Organization physical activity guidelines recommend adults and older adults to accumulate at least 150-300 min of moderate or 75-150 min of vigorous aerobic-type physical activity weekly for health benefits including improvements of cognitive performance. However, the optimal exercise intensity and frequency for maximizing the cognitive benefits remain unclear. Purpose: We conducted a parallel, assessor-blinded, pilot randomized controlled trial to evaluate the effectiveness of different intensities and frequencies of the WHO-recommended minimal volume of aerobic-type physical activity on improving cognitive performance in middle-aged and older adults with mild cognitive impairment (MCI). Methods: Participants were randomly allocated to the stretching exercise control group (CON), once-a-week and thrice-a-week moderate-intensity walking groups (M1 and M3), and once-a-week and thrice-a-week vigorous-intensity walking groups (V1 and V3). Intervention duration was 12 weeks. The primary outcome was global cognitive performance assessed by the Hong Kong version of Montreal Cognitive Assessment. Secondary outcomes were self-report and objective cognitive performances, mental health, sleep quality, and cardiorespiratory fitness. Results: Thirty-seven participants completed the study (CON: n = 7, M1: n = 7, M3: n = 7, V1: n = 8, V3: n = 8). Participants in all four walking exercise groups demonstrated significant improvements in global cognitive performance assessed by the Hong Kong version of the Montreal Cognitive Assessment after the intervention when compared to CON (p < 0.001). The walking exercise interventions also significantly mitigated the anxiety severity (p < 0.005) and improved the cardiorespiratory fitness (p < 0.05) of the participants in the walking exercise groups. Conclusion: 150-min moderate- or 75-min vigorous-intensity walking exercise performed once- or thrice-weekly showed similar effects on improving cognitive performance in middle-aged and older adults with MCI. The 12-week walking exercise interventions also reduced anxiety severity and improved cardiorespiratory fitness of the participants. Clinical Trial Registration: clinicaltrials.gov, identifier NCT04515563.

Can ten days of heat acclimation training improve temperate-condition rowing performance in national-level rowers?

This study investigated whether heat acclimation (HA) could improve rowing performance in temperate conditions in national-level rowers. Using a parallel-group design, eleven rowers (3 female, 8 male, age: 21±3 years, height: 182.3±6.8cm, mass: 79.2±9.0kg, [Formula: see text]: 61.4±5.1ml·kg·min-1) completed either a HA intervention (HEAT, n = 5) or acted as controls (CON, n = 6). The intervention replaced usual cross-training sessions and consisted of an hour of submaximal cycling or rowing ergometry in either 34±0°C for HEAT or 14±1°C for CON daily over two five-day blocks (10 sessions total), separated by 72h. Participants performed the '10+4' test that consists of 10-min submaximal rowing and a 4-min time-trial (TT) in temperate conditions (20±0°C) before and after the intervention. Heat acclimation following the 10-session intervention was evidenced by large significant (p<0.05) decreases in maximum tympanic temperature (d = -1.68) and rate of perceived exertion (RPE) (d = -2.26), and a large significant increase in sweat loss (d = 0.91). Large non-significant (p>0.05) decreases were seen in average tympanic temperature (d = -3.08) and average heart rate (d = -1.53) in HEAT from session 2 to session 10 of the intervention. Furthermore, a large significant increase was seen in plasma volume (d = 3.74), with large significant decreases in haemoglobin concentration (d = -1.78) and hematocrit (d = -12.9). Following the intervention, large non-significant increases in respiratory exchange ratio (d = 0.87) and blood lactate (d = 1.40) as well as a large non-significant decrease in RPE (d = -1.23) were seen in HEAT during the 10-min submaximal rowing. A large significant decrease in peak heart rate (d = -2.27), as well as a large non-significant decrease in relative [Formula: see text] (d = -0.90) and large non-significant increases in respiratory exchange ratio (d = 1.18), blood lactate concentration (d = 1.25) and power output (d = 0.96) were seen in HEAT during the 4-min TT. This study suggests that a 10-session HA intervention may elicit HA in national-level rowers, with potential to improve 4-min TT performance in temperate conditions.

Comparative effectiveness of exercise, antidepressants and their combination in treating non-severe depression: a systematic review and network meta-analysis of randomised controlled trials.

OBJECTIVE: To assess the comparative effectiveness of exercise, antidepressants and their combination for alleviating depressive symptoms in adults with non-severe depression. DESIGN: Systematic review and network meta-analysis. DATA SOURCES: Embase, MEDLINE, PsycINFO, Cochrane Library, Web of Science, Scopus and SportDiscus. ELIGIBILITY CRITERIA: Randomised controlled trials (1990-present) that examined the effectiveness of an exercise, antidepressant or combination intervention against either treatment alone or a control/placebo condition in adults with non-severe depression. STUDY SELECTION AND ANALYSIS: Risk of bias, indirectness and the overall confidence in the network were assessed by two independent investigators. A frequentist network meta-analysis was performed to examine postintervention differences in depressive symptom severity between groups. Intervention drop-out was assessed as a measure of treatment acceptability. RESULTS: Twenty-one randomised controlled trials (n=2551) with 25 comparisons were included in the network. There were no differences in treatment effectiveness among the three main interventions (exercise vs antidepressants: standardised mean differences, SMD, -0.12; 95% CI -0.33 to 0.10, combination versus exercise: SMD, 0.00; 95% CI -0.33 to 0.33, combination vs antidepressants: SMD, -0.12; 95% CI -0.40 to 0.16), although all treatments were more beneficial than controls. Exercise interventions had higher drop-out rates than antidepressant interventions (risk ratio 1.31; 95% CI 1.09 to 1.57). Heterogeneity in the network was moderate (τ2=0.03; I2=46%). CONCLUSIONS: The results suggest no difference between exercise and pharmacological interventions in reducing depressive symptoms in adults with non-severe depression. These findings support the adoption of exercise as an alternative or adjuvant treatment for non-severe depression in adults. SYSTEMATIC REVIEW REGISTRATION: PROSPERO CRD4202122656.

Emergence of a mutation in the nucleocapsid gene of SARS-CoV-2 interferes with PCR detection in Canada.

The emergence of Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV-2) was met with rapid development of robust molecular-based detection assays. Many SARS-CoV-2 molecular tests target multiple genetic regions of the virus to maximize detection and protect against diagnostic escape. Despite the relatively moderate mutational rate of SARS-CoV-2, numerous mutations with known negative impact on diagnostic assays have been identified. In early 2021, we identified four samples positive for SARS-CoV-2 with a nucleocapsid (N) gene drop out on Cepheid Xpert® Xpress SARS-CoV-2 assay. Sequencing revealed a single common mutation in the N gene C29200T. Spatiotemporal analysis showed that the mutation was found in at least six different Canadian provinces from May 2020 until May 2021. Phylogenetic analysis showed that this mutation arose multiple times in Canadian samples and is present in six different variants of interest and of concern. The Cepheid testing platform is commonly used in Canada including in remote regions. As such, the existence of N gene mutation dropouts required further investigation. While commercial SARS-CoV-2 molecular detection assays have contributed immensely to the response effort, many vendors are reluctant to make primer/probe sequences publicly available. Proprietary primer/probe sequences create diagnostic 'blind spots' for global SARS-CoV-2 sequence monitoring and limits the ability to detect and track the presence and prevalence of diagnostic escape mutations. We hope that our industry partners will seriously consider making primer/probe sequences available, so that diagnostic escape mutants can be identified promptly and responded to appropriately to maintain diagnostic accuracy.