Disease Severity Research Articles

Current understanding and future prospects for ash dieback disease with a focus on Britain

Abstract Hymenoscyphus fraxineus is an introduced ascomycete fungus which causes ash dieback and has resulted in widespread mortality of ash throughout Europe. Although H. fraxineus has been present on the continent for at least four decades, it was not identified until 2006. The first record of the pathogen in Britain came in 2012 although it was probably present a decade earlier. The most common host European ash (Fraxinus excelsior L.) is economically and ecologically important to Britain where the cost of ash dieback is estimated at billions of pounds. The impact of ash dieback has stimulated a major research response which we review with the aim of providing up-to-date information relevant to Britain and identifying knowledge gaps where research would contribute to improved disease mitigation. Hymenoscyphus fraxineus is an outcrossing fungus with high genotypic diversity; ascospores produced via sexual reproduction are critical to aerial dispersal and infection. Temperature, moisture, and ground cover influence pathogen fruit body development, the timing of ascospore release, and extent of ascospore germination; they also interact together to affect the likelihood of infection. In addition, stand characteristics, including tree density, tree height, and landscape fragmentation, affect disease dynamics with increased disease severity on moist sites with high ash density. Efforts at finding natural resistance in ash have identified genetic markers associated with disease tolerance, and gene expression analysis is providing insights into the basis of that resistance. Mainland European findings indicate that ash dieback makes trees more vulnerable to other pathogens, whilst endophytes in the ash phyllosphere can suppress infection by H. fraxineus. Possible tools for long-term control of ash dieback include (1) deployment of resistant ash, (2) quantitatively informed management practices based on microclimate models and better understanding of the pathogen life cycle, and (3) manipulation of biocontrol agents from the ash microbiome or pathogen mycoviruses.

Pattern of Expression of Genes Involved in Systemic Inflammation and Glutathione Metabolism Reveals Exacerbation of COPD

To test the hypothesis that they serve as systemic biomarkers of chronic obstructive pulmonary disease (COPD), we profiled the mRNA expression of enzymes connected to systemic inflammation and GSH metabolism in peripheral blood mononuclear cells (PBMCs). These were taken from patients displaying acute exacerbation of COPD (AE-COPD) and stable COPD, and also from non-obstructive smokers and non-smokers. The expression of poly(ADP-ribose) polymerase-1 was increased, but that of histone deacetylase 2 was decreased in association with AE-COPD. The expression of modulatory subunit of glutamyl–cysteine ligase was higher and that of its catalytic subunit, together with the expression of dipeptidyl peptidase 4, was lower in COPD patients compared with non-obstructive smokers and non-smokers. Leukotriene A4 hydrolase saw increased expression in patients with COPD according to disease severity compared to non-obstructive individuals, whereas the expression of GSH peroxidase increased in non-obstructive smokers and COPD patients with the growing number of pack-years smoked. The results corroborate COPD and its acute exacerbation as a complex systemic disorder demonstrating distinct associations with the expression of enzymes linked to inflammation and the regulation of GSH metabolism.

Determinant of Bronchoarterial Ratio in a Patient Undergoing Computed Tomography Chest

Background: Respiratory diseases pose a significant challenge in Nepali clinical settings due to elevated smoking rates and environmental risks. This study explores the structural changes in conditions such as asthma, chronic obstructive pulmonary disease, and bronchiectasis, utilizing noninvasive multi-detector computed tomography to assess airway remodeling. Despite conflicting western literature on bronchoarterial ratios, this research emphasizes the importance of evaluating these ratios in the Nepalese context for predicting airway disease severity. Materials and Methods: The cross-sectional study conducted at Nobel Medical College and Teaching Hospital involved 129 participants aged over 20 undergoing chest computed tomography. It aimed to measure the bronchoarterial ratio and investigate its associations with age, gender, smoking history, and altitude of residence. Bronchoarterial ratios was calculated using averaged short-axis diameters of the right apical and right posterior basal segmental bronchi, excluding the left lung due to motion artifacts. Results: Analysis indicated a Mean Bronchoarterial Ratio of 0.823 (SD 0.151), with the apical and basal segment BARs at 0.832 (SD 0.148) and 0.814 (SD 0.153), respectively. Notably, individuals aged over 65 who were smokers exhibited a significant increase in BAR. Additionally, a higher altitude of residence demonstrated a significant correlation with the Bronchoarterial ratio in the apical lung segment. Conclusion: This study provides valuable insights into the bronchoarterial ratio within the Nepalese population, considering age, gender, smoking history, and altitude of residence. The findings underscore the potential relevance of these ratios in conditions involving airway remodeling, contributing to the understanding of respiratory diseases in this specific demographic.

Automated Detection of Plant Diseases and Pests Using Advanced Image Processing and Neural Networks

Pests and viral plant diseases have proliferated in agriculture. Rats spread plant diseases. Hard to detect viral infections or insect infestations that impair plant growth and yield. It decreases crop size, quality, and marketability, incurring major economic losses. Strong, disease-resistant crops are needed for agriculture. Bacterial, fungal, and insect/pest diseases destroy 70% and 15% of crops. Above 85% of disease severity destroys crops. Early detection systems track pests and diseases. High-definition equipment is costly for rural farmers. Correct diagnosis permits quick infection severity and type assessment and control. In precision agriculture, neural networks and image processing eliminate human error. Sustainable agriculture and organic agricultural cultivation must start early. The proposed system classifies diseases and pests for plant safety. A healthy, fast-growing plantation is our goal. Early detection of viral diseases and insect infestations boosts crop productivity. Plant disease diagnostics focuses on leaves and buds. Stem and insect infections are covered. The suggested method classifies plant health and disease automatically. Peanut leaves develop Bacterial Blight from contagious illnesses like Cercospora Leaf Peanut. Boll Weevils, European Corn Borers, and Fall Army Worms attack cotton. The conditions are categorised. Image accuracy depends on lighting, resolution, location, and backdrop complexity. Various low-resolution pictures are enriched to better classification. Complex leaf intensity variations cause the system to misdiagnose illness or insect infestation. To avoid misclassification, the system segments the sample image using various methods to find lesions. Multilevel Segmentation (MS) improves Deep neural network feature extraction using approximate outlines. This method protects important pixels, eliminates unneeded pixels, and amplifies the lesion to control segmentation and reduce insufficient and excessive segmentation.

Chromatin Regulator SMARCA4 Is Essential for MHV-Induced Inflammatory Cell Death, PANoptosis

The innate immune system serves as the first line of defense against β-coronaviruses (β-CoVs), a family of viruses that includes SARS-CoV-2. Viral sensing via pattern recognition receptors triggers inflammation and cell death, which are essential components of the innate immune response that facilitate viral clearance. However, excessive activation of the innate immune system and inflammatory cell death can result in uncontrolled release of proinflammatory cytokines, resulting in cytokine storm and pathology. PANoptosis, innate immune, inflammatory cell death initiated by innate immune sensors and driven by caspases and RIPKs through PANoptosome complexes, has been implicated in the pathology of viral infections. Therefore, understanding the molecular mechanisms regulating PANoptosis in response to β-CoV infection is critical for identifying new therapeutic targets that can mitigate disease severity. In the current study, we analyzed findings from a cell death-based CRISPR screen with archetypal β-CoV mouse hepatitis virus (MHV) as the trigger to characterize host molecules required for inflammatory cell death. As a result, we identified SMARCA4, a chromatin regulator, as a putative host factor required for PANoptosis in response to MHV. Furthermore, we observed that gRNA-mediated deletion of Smarca4 inhibited MHV-induced PANoptotic cell death in macrophages. These findings have potential translational and clinical implications for the advancement of treatment strategies for β-CoVs and other infections.

IL-10+ regulatory B cells mitigate atopic dermatitis by suppressing eosinophil activation

Atopic dermatitis (AD) presents significant therapeutic challenges due to its poorly understood etiology. Eosinophilia, a hallmark of allergic inflammation, is implicated in AD pathogenesis. Interleukin-10 (IL-10)-producing regulatory B (Breg) cells exhibit potent anti-inflammatory effects. However, their role in controlling AD-related eosinophilia is not well understood. To investigate the impact of eosinophils on AD, we employed IL-5Rα-deficient (Il5ra−/−) mice, which lack functional eosinophils. Induction of AD in these mice resulted in attenuated disease symptoms, underscoring the critical role of eosinophils in AD development. Additionally, the adoptive transfer of purified Breg cells into mice with AD significantly alleviated disease severity. Mechanistic studies revealed that IL-10 produced by Breg cells directly inhibits eosinophil activation and infiltration into the skin. In vitro experiments further confirmed that Breg cells inhibited eosinophil peroxidase secretion in an IL-10-dependent manner. Our collective findings demonstrate that IL-10 from Breg cells alleviates AD by suppressing eosinophil activation and tissue infiltration. This study elucidates a novel regulatory mechanism of Breg cells, providing a foundation for future Breg-mediated therapeutic strategies for AD.

Mechanobiological factors in fermentation: Understanding the impact on food texture and nutrient release

Evaluating people’s food intake is crucial for establishing the link between diet and disease. Deprivation of vital nutrients causes the body to deteriorate organs and increases the risk of severe diseases that manifest in maturity. Making healthy food choices is one of the best ways to avoid developing chronic diseases, such as diabetes, heart disease, stroke, and even certain types of cancer. Hence, this paper proposes a deep learning-based automated nutrition classification system (DL-ANCS) for predicting food ingredients and nutrition. The DL-ANCS with Internet of Things (IoT) sensors to quantify food nutrition and a smartphone app to compile ingredient nutritional information. It is possible to take pictures of food eaten using the camera that comes with most mobile phones. It is possible to automatically identify the food items for record keeping using image processing. The effectiveness of the proposed DL-ANCS relies on its ability to accurately classify food items in these photos utilizing meal prediction algorithms and DL-ANCS. This research introduces a novel approach to extracting texture information from food photos to show how these features improve the accuracy of a nutritional assessment system that runs on mobile phones. The proposed method improvesthe food texture ratio of 98.7%, effectiveness ratio of 99.2%, accuracy ratio of 95.89%, food ingredient predictionsand their nutritional compatibility ratio of 96.8%, and food component classifications ratio of 97.29%.

Serologically Confirmed Human Leptospirosis in Colombia, 2015-2020.

Leptospirosis, a bacterial infection transmitted through contact with infected animals or contaminated water sources, imposes a substantial health burden in Colombia. Since 2007, the National Institute of Health (INS) has mandated the notification and confirmation of all suspected leptospirosis cases. This passive surveillance program employs the microscopic agglutination test (MAT) on serum samples to ascertain confirmed cases of leptospirosis infection. However, the absence of a robust surveillance system has hindered our comprehensive understanding of the morbidity, mortality, geographical distribution, species/serovars, and strains responsible for severe disease. Our study aimed to provide an epidemiological overview of MAT-confirmed human leptospirosis cases reported over 6 years (2015-2020) in Colombia. In addition, we offer insights into the status of leptospirosis in the country, focusing on risk factors and proposing potential improvements for diagnosis and disease management. During the 6-year surveillance period, the laboratory at the INS received 3,535 serum samples from suspected human leptospirosis cases, with 880 (25%) confirmed through MAT. The incidence of leptospirosis was calculated at 1.9 cases per 100,000 people, with a higher prevalence among men (82.1%). Furthermore, 54 (6.1%) deaths were confirmed as leptospirosis, and cases were documented across nearly all regions of Colombia. Our findings emphasize the urgent need to strengthen leptospirosis laboratory surveillance, implement effective prevention measures, and enhance diagnostic capabilities in Colombia. The analysis conducted in this study provides the groundwork for estimating the impact of leptospirosis and raises awareness of its significance in public health.

Serum inflammatory markers in children with Mycoplasma pneumoniae pneumonia and their predictive value for mycoplasma severity.

Mycoplasma pneumoniae pneumonia (MPP) significantly impacts pediatric health, necessitating markers for early severe disease identification. To investigate the correlation between serum inflammatory marker and the severity of MPP in children. A prospective study was carried out from January 2023 to November 2023. A total of 160 children with MPP who underwent treatment were selected: 80 had severe MPP and 80 had mild MPP. Clinical and laboratory data were collected at the time of hospital admission and during hospitalization. Receiver operating characteristic curves were utilized to assess the diagnostic and prognostic for severe MPP. Fever duration and length of hospitalization in pediatric patients with severe MPP exceeded those with mild MPP. The incidence of pleural effusion, lung consolidation, and bronchopneumonia on imaging was markedly elevated in the severe MPP cohort compared to the mild MPP cohort. In contrast to the mild cohort, there was a notable increase in C-reactive protein (CRP), procalcitonin (PCT), erythrocyte sedimentation rate, lactic dehydrogenase, D-dimer, and inflammatory cytokines [interleukin (IL)-6, IL-8, IL-10 and tumor necrosis factor (TNF)-α] in the severe MPP group were significantly higher. Serum inflammatory markers (CRP, PCT, IL-6, D-dimer, IL-10 and TNF-α) were considered as predictors in children with severe MPP.

Below-the-Ankle Antegrade Teleferic Technique: New Approach for Below-the-Ankle Revascularization in Challenging Cases of Chronic Limb-Threatening Ischemia.

To describe a novel bailout technique to approach below-the-ankle (BTA) chronic total occlusions or plantar-arch severe disease where the balloon/catheter is unable to follow the crossing guidewire and no other described recanalization approach is feasible. When facing a complex BTA revascularization, if the guidewire crosses but the balloon cannot progress due to a lack of pushability, an antegrade puncture of the infrapopliteal vessel where the tip of the guidewire lays is performed. The guidewire is then carefully navigated through this distal BTA vessel into the needle to achieve its rendezvous and externalization. A low-profile balloon is inserted through the femoral access and advanced till the non-crossable point of the BTA vessels. A torque device is then attached to the proximal hub of the balloon, and the through-and-through guidewire is subsequently pulled from the new distal access, allowing the balloon to be dragged across the lesion together with the wire. The below-the-ankle antegrade teleferic (BAT) technique may be considered for highly complex BTA revascularization procedures where the wire crosses the lesion, but no other device can be tracked over it. The clinical impact of this article lies in the description of a bailout technique for BTA revascularization where the guidewire crosses, but no device can be advanced. This technique can be helpful in scenarios where failure to achieve success could result in limb loss. The BAT technique provides a solution in extremely challenging cases, enhancing technical success, improving outcomes and potentially preserving the limbs of patients who would otherwise face amputation, if not revascularized.The video shows the BAT technique performed with a support catheter under fluoroscopy: antegrate puncture of the DP, advancement of the support catheter over the wire, rendezvous of the guidewire in the catheter and subsequent externalization of the wire.

The Dynamics of NS1 Antigen and Platelet Levels in the Acute Phase of Dengue Hemorrhagic Fever

Introduction: Dengue hemorrhagic fever (DHF) is a significant global health concern, particularly in tropical and subtropical regions. The early diagnosis and effective management of DHF are crucial to reduce morbidity and mortality. The NS1 antigen and platelet count are two key laboratory parameters used in the diagnosis and monitoring of DHF. This study aimed to investigate the dynamics of NS1 antigen and platelet levels during the acute phase of DHF and their correlation with disease severity. Methods: A prospective cohort study was conducted at a tertiary care hospital in Central Java, Indonesia. Patients presenting with suspected DHF were enrolled and followed up during their hospitalization. Blood samples were collected at admission and at regular intervals to measure NS1 antigen levels and platelet counts. Clinical and laboratory data were collected to assess disease severity. Results: A total of 150 patients were included in the study. The majority of patients were adults (median age, 32 years) with a female predominance (58%). The median duration of fever at presentation was 4 days. NS1 antigen levels were significantly higher in patients with DHF compared to those with dengue fever (DF) (p < 0.001). Platelet counts were significantly lower in patients with DHF compared to those with DF (p < 0.001). NS1 antigen levels showed a negative correlation with platelet counts (r = -0.62, p < 0.001). The levels of NS1 antigen and platelet count were also significantly associated with disease severity. Conclusion: The dynamics of NS1 antigen and platelet levels during the acute phase of DHF provide valuable insights into disease pathogenesis and can aid in the early diagnosis and prediction of disease severity. The negative correlation between NS1 antigen and platelet levels suggests a potential role of NS1 in the development of thrombocytopenia in DHF. Further studies are needed to elucidate the exact mechanisms underlying these observations.

The Circadian Rhythm of Itching among 241 Adults with Atopic Dermatitis: A Cross-sectional Study.

The pattern of itching in patients with atopic dermatitis has not been systematically studied. Therefore, this study aimed to assess the pattern of itching in adults with atopic dermatitis using questionnaires to assess for a circadian rhythm of itching in participating patients at a single institution (n = 241). A self-report questionnaire was used to assess circadian rhythm and intensity of itching in patients. In addition, the patients' disease severity (Eczema Area and Severity Index [EASI]) and quality of life (Dermatology Life Quality Index [DLQI]) were assessed. Itching occurred most frequently (74.69%) and with the greatest severity (62.66%) between 20:00 and 00:00, and the least number of patients (25.31%) experienced itching between 04:00 and 08:00. The DLQI and EASI scores both correlated with the average and maximum itch intensity (r = 0.582, r = 0.533, respectively; r = 0.539, r = 0.517, respectively; p < 0.001). The DLQI and EASI scores were associated with average itch intensity (B = 0.179, B = 0.204, respectively; 95% CI: 0.112 to 0.246, 95% CI: 0.096 to 0.313, respectively; p < 0.001), and the EASI score was associated with males and family history (B = 0.285, B = 0.287, respectively; 95% CI: 0.094 to 0.476, 95% CI: 0.096 to 0.478, respectively; p = 0.003). Adult patients with atopic dermatitis exhibited a circadian rhythm of itching; these study results could positively impact treatment approaches.

Adjunctive treatment of atopic dermatitis with novel at-home handheld narrow-band UVB phototherapy.

While conventional in-office phototherapy has long been utilized as a successful treatment for atopic dermatitis (AD), it is associated with potential barriers including inconvenience, poor adherence, time and financial expense. In this retrospective study, we examine the efficacy, adherence, and patient-satisfaction of using adjunctive at-home, self-administered phototherapy utilizing a novel handheld narrow-band ultraviolet B (NB-UVB) device for the treatment of refractory mild to severe AD. Included AD patients were initially trained on proper use of the device. These patients treated involved areas three times per week for a period of 12weeks. Phototherapy dosing protocol was based on skin type. The cohort included 52 patients, who were aged 20-69 and represented all skin types. They were initially categorized by disease involvement as mild, moderate, and severe. Patients were also queried to self-score their disease severity and level of satisfaction. Compared to baseline, at 12weeks, 48% percent of patients indicated that at least one site was Clear/Almost Clear, 38% stated that more than 50% of body locations were Clear/Almost Clear, and 28% reported that 100% (all) treated sites were Clear/Almost Clear. After using at-home hand-held NB-UVB for the study duration, 67% (35/52) of patients experienced disease improvement. Mean overall satisfaction was extremely high at 4.43 on a 5-point scale. Skin type, age, gender, and disease severity at inception did not significantly affect patient satisfaction scores. Overall adherence rate among participants across all groups was 73%. In this small retrospective study, at-home handheld NB-UVB phototherapy was found to be an effective, well-tolerated, adjunctive treatment method for patients with refractory AD, which was associated with a high level of patient satisfaction and adherence.

Frailty and sarcopenia in patients with cirrhosis awaiting liver transplantation: evidence from a single-centre, prospective cohort study.

Sarcopenia and frailty are common complications in patients with cirrhosis evaluated for liver transplantation (LT). Although the negative impact of sarcopenia on patient's outcome has been well studied, the prognostic role of frailty is not as clear. We assessed the prevalence of sarcopenia and frailty and the clinical impact of frailty in a prospective cohort of cirrhosis patients with and without hepatocellular carcinoma (HCC) listed for LT. Patients with cirrhosis were prospectively recruited at the time of admission into the waiting list. Clinical and lab values were collected. Physical frailty was assessed by liver frailty index (LFI) and patients were categorized into robust (< 3.2); pre-frail (between 3.2 and 4.5), and frail (> 4.5). Skeletal muscle mass was evaluated via skeletal muscle index (SMI) obtained from last CT scan before LT; sarcopenia was defined by SMI < 50 cm2/m2 in males and < 39 cm2/m2 in females. 105 patients were included, of which 42 (40%) had hepatocellular carcinoma (HCC). In patients without HCC (63.5% males, median age 61 years), 36.5% were frail, 50.8% were pre-frail and 12.7% were robust. Frail patients were older than non-frail patients (63 vs. 56; p = 0.008) and had more severe liver disease (Child C: 65% vs. 37.5%; p = 0.02). Prevalence of sarcopenia in patients without HCC was 63%, with similar value of median SMI between frail and not frail patients (p = 0.454). Patients with HCC (78.6% males, 65years old) were 21.4% frail, 61.9% pre-frail, and 16.7% robust. Frail patients had more severe liver disease (Child C: 77% vs. 18.2%; p = 0.004), whereas age was comparable to non-frail patients; among patients without HCC, during a median follow-up of 263days, 17% died (of which 72% were frail) and 10 patients were delisted due to clinical improvement (none of whom were frail). Among those with HCC, during a median follow-up of 289 days, 4 (9%) patients died of which 50% were frail. Frailty and sarcopenia are common complications in patients with cirrhosis awaiting LT. Frailty appears to be associated with an increased risk of mortality during wait-list time especially in those with decompensated cirrhosis. At univariate analysis Meld score, Child score and presence of frailty were found to be associated with shorter survival, however, at multivariate analysis presence of frailty and Child C vs. A/B were the only independent predictor of death. Larger cohorts are required to confirm these results.

Assessment of acute neuronal injury in critical illness: prognostication in septic shock patients - preliminary study in a Polish population.

Sepsis-associated brain dysfunction is a common organ dysfunction in sepsis. The main goal of this study was to verify whether the combined assessment of central nervous system injury markers (i.e. S100B, NSE, GFAP) and disease severity as per the Acute Physiology and Chronic Health Evaluation II (APACHE II), Simplified Acute Physiology Score II (SAPS II), and Sequential Organ Failure Assessment (SOFA) classification systems, would increase the accuracy of death prediction in septic shock. Markers of neuronal damage were determined in 55 patients diagnosed with septic shock with no previous neurological disease. Clinical data was collected and the scores on the APACHE II, SAPS II and SOFA prognostic scales were calculated. Death before discharge from the Intensive Care Unit (ICU) was established as the endpoint. Nineteen patients (35%) died before ICU discharge. Patients who died had significantly higher S100B and NSE values, and APACHE II, SAPS II and SOFA scores (P< 0.05 for all). At the time of septic shock diagnosis, NSE levels more accurately predicted the risk of death before ICU discharge than S100B. However, NSE had no better predictive value for short-term mortality than APACHE II, SAPS II and SOFA. Adding C-reactive protein (CRP) and S100B concentrations to the APACHE II score created a predictive model with 95% mortality accuracy (AUC = 0.95; 95%CI 0.85-0.99; P = 0.03). The assessment of acute neuronal injury plays an important role in prognostication in patients with septic shock. The concentration of S100B protein in combination with APACHE II score and concentration of CRP more accurately predicts mortality than the APACHE II alone.

Effect of Weather Parameters on Disease Severity and Progression of Fusarium Wilt in Chilli (Capsicum anuum L.)

Fusarium wilt incited by Fusarium oxysporum is an economically damaging disease of chilli (Capsicum anuum L.). Field experiments on epidemiological studies were conducted using susceptible variety (Pusa Jwala) to study the effect of weather parameters on the progression of the disease. The study revealed that transplanting chilli during the first half of July reduced the disease severity compared to earliest transplant dates. The area under disease progress curve (AUDPC) and the apparent infection rate were lowest in the crop transplanted on 15th July, 2022 while they were highest in the crop transplanted on 15th June, 2022. The correlation analysis showed a positive correlation between disease severity and bright sunshine hours, and a negative correlation with the other weather parameters. The R2 value obtained from multiple regression equations revealed that the combined effect of weather parameters could explain the prediction of a disease level of 83-86 per cent at all three transplant dates.

Rapid Detection of Viral, Bacterial, Fungal, and Oomycete Pathogens on Tomatoes with Microneedles, LAMP on a Microfluidic Chip, and Smartphone Device.

Rapid detection of plant diseases before they escalate can improve disease control. Our team has developed rapid nucleic acid extraction methods with microneedles and combined these with loop-mediated amplification (LAMP) assays for pathogen detection in the field. In this work, we developed LAMP assays for early blight (Alternaria linariae, A. alternata, and A. solani) and bacterial spot of tomato (Xanthomonas perforans) and validated these LAMP assays and two previously developed LAMP assays for tomato spotted wilt virus and late blight. Tomato plants were inoculated, and disease severity was measured. Extractions were performed using microneedles, and LAMP assays were run in tubes (with hydroxynaphthol blue) on a heat block or on a newly designed microfluidic slide chip on a heat block or a slide heater. Fluorescence on the microfluidic chip slides was visualized using EvaGreen and photographed on a smartphone. Plants inoculated with X. perforans or tomato spotted wilt virus tested positive prior to visible disease symptoms, whereas Phytophthora infestans and A. linariae were detected at the time of visual disease symptoms. LAMP assays were more sensitive than PCR, and the limit of detection was 1 pg of DNA for both A. linariae and X. perforans. The LAMP assay designed for early blight detected all three species of Alternaria that infect tomato and is thus an Alternaria spp. assay. This study demonstrates the utility of rapid microneedle extraction followed by LAMP on a microfluidic chip for rapid diagnosis of four important tomato pathogens.

Fluoxetine accelerates epileptogenesis and magnifies disease severity in a rat model of acquired epilepsy.

Many people with epilepsy experience comorbid anxiety and depression, and antidepressants remain a primary treatment for this. Emerging evidence suggests that these agents may modulate epileptogenesis to influence disease severity. Here, we assessed how treatment with the selective serotonin reuptake inhibitor (SSRI) antidepressant fluoxetine impacts epileptogenic, behavioral, and pathological sequelae following status epilepticus. Male Wistar rats received kainic acid to induce status epilepticus (SE) or vehicle (sham). Animals then received either fluoxetine (10 mg/kg/day) or vehicle for 8 weeks via subcutaneous osmotic pump. Video-electroencephalography was recorded continuously until behavioral testing at day 56, including assessments of anxiety- and depression-like behavior and spatial cognition. Postmortem immunocytochemistry studies examined mossy fiber sprouting. Fluoxetine treatment significantly accelerated epileptogenesis following SE, reducing the average period to the first spontaneous seizure (from 32 days [vehicle] to 6 days [fluoxetine], p < .01). Also, fluoxetine exposure magnified the severity of the resultant epilepsy, increasing seizure frequency compared to vehicle (p < .01). Exposure to fluoxetine was associated with improved anxiety- and depression-like behaviors but significantly worsened cognition. Mossy fiber sprouting was more pronounced in fluoxetine-treated rats compared to vehicle (p < .0001). Our studies demonstrate that, using a model exhibiting spontaneous seizures, epileptogenesis is accelerated and magnified by fluoxetine, an effect that may be related to more severe pathological neuroplasticity. The differential influence of fluoxetine on behavior indicates that different circuitry and mechanisms are responsible for these comorbidities. These findings suggest that caution should be exercised when prescribing SSRI antidepressants to people at risk of developing epilepsy.

Contemporary Practice Patterns in the Treatment of Cervical Stenosis and Central Cord Syndrome: A Survey of the Cervical Spine Research Society.

Cross-sectional study. To evaluate for areas of consensus and divergence of opinion within the spine community regarding the management of cervical spondylotic conditions and acute traumatic central cord syndrome (ATCCS) and the influence of the patient's age, disease severity, and myelomalacia. There is ongoing disagreement regarding the indications for, and urgency of, operative intervention in patients with mild degenerative myelopathy, moderate to severe radiculopathy, isolated axial symptomatology with evidence of spinal cord compression, and ATCCS without myelomalacia. A survey request was sent to 330 attendees of the Cervical Spine Research Society (CSRS) 2021 Annual Meeting to assess practice patterns regarding the treatment of cervical stenosis, myelopathy, radiculopathy, and ATCCS in 16 unique clinical vignettes with associated MRIs. Operative versus nonoperative treatment consensus was defined by a management option selected by >80% of survey participants. Overall, 116 meeting attendees completed the survey. Consensus supported nonoperative management for elderly patients with axial neck pain and adults with axial neck pain without myelomalacia. Operative management was indicated for adult patients with mild myelopathy and myelomalacia, adult patients with severe radiculopathy, elderly patients with severe radiculopathy and myelomalacia, and elderly ATCCS patients with pre-existing myelopathic symptoms. Treatment discrepancy in favor of nonoperative management was found for adult patients with isolated axial symptomatology and myelomalacia. Treatment discrepancy favored operative management for elderly patients with mild myelopathy, adult patients with mild myelopathy without myelomalacia, elderly patients with severe radiculopathy without myelomalacia, and elderly ATCCS patients without preceding symptoms. Although there is uncertainty regarding the treatment of mild myelopathy, operative intervention was favored for nonelderly patients with evidence of myelomalacia or radiculopathy and for elderly patients with ATCCS, especially if pre-injury myelopathic symptoms were present. Level V.

Intrinsic link between PGRN and Gba1 D409V mutation dosage in potentiating Gaucher disease.

Gaucher disease (GD) is caused by biallelic GBA1/Gba1 mutations that encode defective glucocerebrosidase (GCase). Progranulin (PGRN, encoded by GRN/Grn) is a modifier of GCase, but the interplay between PGRN and GCase, specifically GBA1/Gba1 mutations, contributing to GD severity is unclear. Mouse models were developed with various dosages of Gba1 D409V mutation against the PGRN deficiency (Grn-/-) [Grn-/-;Gba1D409V/WT (PG9Vwt), Grn-/-;Gba1D409V/D409V (PG9V), Grn-/-;Gba1D409V/Null (PG9VN)]. Disease progression in those mouse models was characterized by biochemical, pathological, transcriptomic, and neurobehavioral analyses. Compared to PG9Vwt, Grn-/-;Gba1WT/Null and Grn-/- mice that had a higher level of GCase activity and undetectable pathologies, homozygous or hemizygous D409V in PG9V or PG9VN, respectively, resulted in profound inflammation and neurodegeneration. PG9VN mice exhibited much earlier onset, shorter life span, tissue fibrosis, and more severe phenotypes than PG9V mice. Glycosphingolipid accumulation, inflammatory responses, lysosomal-autophagy dysfunction, microgliosis, retinal gliosis, as well as α-Synuclein increases were much more pronounced in PG9VN mice. Neurodegeneration in PG9VN was characterized by activated microglial phagocytosis of impaired neurons and programmed cell death due to necrosis and, possibly, pyroptosis. Brain transcriptomic analyses revealed the intrinsic relationship between D409V dosage, and the degree of altered gene expression related to lysosome dysfunction, microgliosis, and neurodegeneration in GD, suggesting the disease severity is dependent on a GCase activity threshold related to Gba1 D409V dosage and loss of PGRN. These findings contribute to a deeper understanding of GD pathogenesis by elucidating additional underlying mechanisms of interplay between PGRN and Gba1 mutation dosage in modulating GCase function and disease severity in GD and GBA1-associated neurodegenerative diseases.