Important Disease Research Articles

Development of an Effective Method to Evaluate Resistance of Onion (Allium cepa L.) Cultivars to Fusarium Basal Rot

Fusarium basal rot (FBR), caused by the ascomycete fungus <i>Fusarium oxysporum</i>, is an economically important disease of onion worldwide. The most economical and effective way to manage FBR would be the use of FBR resistant onion cultivars. This study was carried out to develop a rapid screening method for resistant onion cultivars in seedling stage. We used the <i>F. oxysporum</i> 19-385 isolate, which causes damping-off in onion seedlings and basal rot in onion bulbs. We optimized broth incubation and medium composition for the production of inoculum, and determined conidial concentration for the preparation of <i>F. oxysporum</i> infected soil. Ten commercial cultivars of onion were evaluated the seedling survival rates and heights by infected soil inoculation methods. As a result, ‘K-force’ was the most resistant cultivar with 97.4% of relative seedling survival rate against the pathogen, whereas ‘Sunpower’ was the most susceptible cultivar with 20.0% of relative seedling survival rate.

The Impact of Covid-19 on Lower Urinary Tract Symptoms (LUTS) in Patients Treated Conservatively for Benign Prostatic Hyperplasia

Benign prostatic hyperplasia (BPH) is a non-malignant urological pathology characterized by a progressive increase in the volume of the prostate gland and one of the important diseases which affect lower urinary tract symptoms (LUTS). Covid-19 is a new viral infection with epidemiological characteristics of a pandemic. The disease is presented as a respiratory, cardiovascular, and neurological late complication. There is limited data that low urinary tract can be affected and aggravate symptoms and quality of life (QoL). Our pilot study describes a male population treated with conservative methods for BPH and the effect of Covid-19 infection on their LUTS and QoL. This was a case-control study between April 2021 and November 2021. The study included 33 patients with a history of Covid-19 infection. The assessment of LUTS was carried out by processing the data obtained from the completed by the patients IPSS questionnaires (the last one before, immediately, and five months after Covid-19). The control group consisted of 50 patients, which for the management of the selection bias, matched according to demographic and clinical features of the group. All patients underwent ultrasound diagnostics giving information about the volume of the prostate gland and the amount of residual urine. Two years after the beginning of the Covid-19 pandemic, we can confidently say that not all characteristics of this type of infection are sufficiently clear. More in-depth studies on Covid-19 as a cause of worsening LUTS and overall progression of BPH could help identify new aspects of infection leading to significant improvements in its treatment.

Molecular Identification and Bioinformatics Analysis of Anaplasma marginale Moonlighting Proteins as Possible Antigenic Targets.

Diseases of veterinary importance, such as bovine Anaplasmosis, cause significant economic losses. Due to this, the study of various proteins of the causal agent Anaplasma marginale has focused on surface proteins. However, a vaccine for this disease is not yet available. To this end, in this work, moonlighting proteins (MLPs) are presented as an alternative approach for the design of immunogens against A. marginale. The proteins of the strain MEX-15-099-01 were analyzed, and its MLPs were identified. Subsequently, four virulence-associated MLP genes were selected and identified using PCR. The proteins were analyzed using a structural homology approach and the collection of B-cell epitopes was predicted for each MLP. Finally, a pair of AmEno peptides were synthesized and the antigenic potential was tested using an iELISA. Our bioinformatics analysis revealed the potential of AmEno, AmGroEl, AmEF-Tu, and AmDnaK proteins as promising candidates for designing immunogens. The PCR allowed the gene sequence identification in the genome of the strain MEX-15-099-01. Notably, AmEno-derived synthetic peptides showed antigenicity in an ELISA. Our study has shed light on the potential use of MLPs for immunogen design, demonstrating the antigenic potential of AmEno.

Pharmacokinetics and safety of topical fluralaner in koalas (Phascolarctos cinereus)

Sarcoptic mange (etiological agent Sarcoptes scabiei) is among the most important parasitic diseases of some marsupial species and has been an emerging disease of koalas, causing welfare and conservation implications. Fluralaner (Bravecto® MSD Animal Health), an ectoparasiticide of the isoxazoline class, has been demonstrated as a long-lasting and efficacious chemotherapeutic agent against sarcoptic mange in multiple mammal species and may also be beneficial for impacted koalas. Here, we evaluated the pharmacokinetics and clinical safety of fluralaner in koalas. Healthy captive individuals were treated topically with 85 mg/kg fluralaner administered to the interscapular epidermis. Following treatment, fluralaner was detected in plasma using ultra-performance liquid chromatography and tandem mass-spectrometry over a 12-week period. The mean maximum plasma concentration (Cmax) was 66.4 ng/mL; mean time was Cmax of 2.71 days; plasma elimination half-life (T1/2) was 30.91 days; and mean residence time (MRT) was 27.38 days. Haematological, blood biochemical, animal husbandry and clinical observations, over the same time period, demonstrated fluralaner was well tolerated. Overall, this research suggests fluralaner is a safe and long-lasting chemotherapeutic agent that may be efficacious against S. scabiei in koalas. Further research focussed on quantifying efficacy in captive and field settings, and across a range of disease severities would be valuable.

The effect of diabetes in the multifaceted relationship between education and cognitive function

BackgroundEducation has been shown to be positively associated with cognitive performance. However, the pathways via lifestyle-related disease through which education is related to cognitive performance have not been sufficiently explored. Diabetes is an important lifestyle-related disease with increasing prevalence worldwide. Low education is associated with an increased risk of developing diabetes, while diabetes may also lead to a deterioration in cognitive performance. This study aims to explore if the associations between education and cognitive function is mediated by the diabetes status among older adults.MethodsThe data utilized in this study were derived from the first two waves of the Dutch Lifelines Cohort Study (2006–2015). The analyzed sample included 26,131 individuals aged 50 years or above at baseline. The baseline assessment included measurements of educational attainment (exposure) and the potential mediator diabetes. The outcome of cognitive function was assessed using age-standardized reaction times from the psychomotor function and attention tasks, as measured by the Cogstate Brief Battery. The Cogstate Brief Battery was only conducted at the follow-up assessment, not at the baseline assessment. Faster reaction times correspond to higher cognitive performance. The study employed linear and logistic regression models, in addition to a causal mediation approach which estimated the average causal mediation effect (ACME).ResultsHigher education was associated with a lower risk of diabetes (b= -0.1976, 95%CI= -0.3354; -0.0597) compared to low or middle education as well as with faster reaction times (b= -0.2023, 95%CI= -0.2246; -0.1798), implying better cognitive function. Diabetes was associated with slower reaction times (b = 0.0617, 95%CI = 0.0162; 0.1072). Most importantly, the mediation approach identified a significant indirect effect of education on cognitive function via the diabetes status (ACME= -0.00061, 95%CI= -0.00142; -0.00011).DiscussionThe findings emphasize the potentially importance of diabetes in explaining the role of education in promoting healthy cognitive function and mitigating the risk of cognitive decline. Early detection and treatment of diabetes may be particularly beneficial for individuals with low or middle levels of education in order to maintain good levels of cognitive function.

Genomic and Pathological Characterization of Acute Hepatopancreatic Necrosis Disease (AHPND)-Associated Natural Mutant Vibrio parahaemolyticus Isolated from Penaeus vannamei Cultured in Korea.

Acute hepatopancreatic necrosis disease (AHPND) is one of the most important diseases in the global shrimp industry. The emergence of mutant AHPND-associated V. parahaemolyticus (VpAHPND) strains has raised concerns regarding potential misdiagnosis and unforeseen pathogenicity. In this study, we report the first emergence of a type II (pirA-, pirB+) natural mutant, VpAHPND (strain 20-082A3), isolated from cultured Penaeus vannamei in Korea. Phenotypic and genetic analyses revealed a close relationship between the mutant strain 20-082A3 and the virulent Korean VpAHPND strain 19-021-D1, which caused an outbreak in 2019. Detailed sequence analysis of AHPND-associated plasmids showed that plasmid pVp_20-082A3B in strain 20-082A3 was almost identical (>99.9%) to that of strain 19-021-D1. Moreover, strains 20-082A3 and 19-021-D1 exhibited the same multilocus sequence type (ST 413) and serotype (O1:Un-typeable K-serogroup), suggesting that the mutant strain is closely related to and may have originated from the virulent strain 19-021-D1. Similar to previous reports on the natural mutant VpAHPND, strain 20-082A3 did not induce AHPND-related symptoms or cause mortality in the shrimp bioassay. The emergence of a mutant strain which is almost identical to the virulent VpAHPND highlights the need for surveillance of the pathogen prevalent in Korea. Further investigations to elucidate the potential relationship between ST 413 and recent Korean VpAHPND isolates are needed.

Suspected Case of Persistent Thiamin Deficiency in a Hand-Reared Caribou Calf.

Hand-reared animals are invaluable and irreplaceable in studies of wildlife nutrition. Hand-rearing protocols provide insights into dietary and training programs, but less information is available on disease management. In young ruminants, thiamin (Vitamin B1) deficiency is a particularly important disease that is treatable early in the disease process, but otherwise can be fatal. In this husbandry report, we describe a case of suspected thiamin deficiency in a hand-reared calf (Rangifer tarandus granti) that resulted in clinical signs of polioencephalomalacia and persisted for > 3 months. We attempted treatment with thiamin injections; injections resolved clinical signs of disease, but clinical signs of disease returned once injections stopped. After > 2 months of thiamin injections, the caribou calf received a rumen transfaunation from a fistulated moose (Alces alces) housed at the same facility. Following rumen transfaunation, we did not observe signs of thiamin deficiency. The calf outgrew other females in the cohort initially and shows no long-term effects of thiamin deficiency or polioencephalomalacia. We recommend rumen transfaunation when thiamin deficiency is suspected and does not resolve with thiamin injections alone. We also recommend heterospecific donors if conspecific donors are not available.

Identification of fungal species associated with gall oak (Quercus infectoria Oliv.) decline in Iran.

The gall oak (Quercus infectoria Oliv.) tree is one of the most important and valuable forestry species in the Northern Zagros forests in the west of Iran. Gall oak decline is considered to be one of the most important diseases currently affecting the oak Zagros forests in Iran. The main objective of the present study, conducted in the years 2021-2023, was to investigate the possible role of fungi as causative agents of gall oak dieback in the Zagros forests of Iran. Wood samples were taken from gall oak trees showing canker, dieback, and internal wood discoloration symptoms. Fungal isolates recovered from gall oak trees were identified based on cultural and morphological characteristics, as well as phylogenetic analyses using DNA sequencing of the internal transcribed spacer region of rDNA (ITS) and partial beta-tubulin (tub2). Achaetomium aegilopis, Alternaria tenuissima, Apiospora intestine, Botrytis cinerea, Coniochaeta sp., Coniothyrium palmarum, Coniothyrium sp., Cytospora rhodophila, Dialonectria episphaeria, Diatrype sp., Diatrypella macrospora, Endoconidioma populi, Fonsecazyma sp., Fusarium ipomoeae, Jattaea discrete, Kalmusia variispora, Microsphaeropsis olivacea, Neoscytalidium dimidiatum, Paecilomyces lecytidis, Paramicrosphaeropsis eriobotryae, Paramicrosphaeropsis ellipsoidea, and Seimatosporium pezizoides were identified from diseased trees. Pathogenicity tests were performed by artificial inoculation of excised branches of healthy gall oak trees under controlled conditions and evaluated after 35 days by measuring the discolored lesion length at the inoculation site. N. dimidiatum was the most virulent species and caused the longest wood necrosis within 35 days of inoculation. In the greenhouse test, only some species induced typical symptoms of canker. All isolated fungi are reported for the first time on gall oak trees in the world.

A qPCR assay for specific detection of Pseudomonas cannabina pv. alisalensis.

Pseudomonas cannabina pv. alisalensis is a gram-negative bacterium that causes bacterial leaf blight in Brassica crops, an important disease that could bring severe damage to the host plants. The aim of this study was to develop a tool that can reliably and accurately quantify P. cannabina pv. alisalensis and distinguish it from other closely related bacterial pathogens. Two species and six pathovars of Pseudomonas were tested: three pathovars, P. syringae pv. coriandricola, P. syringae pv. philadelphi, and P. syringae strains from Vicia faba were found or confirmed to be members of P. cannabina based on the multi-locus sequence analysis and rep-PCR results. The qPCR assay was evaluated for specificity and examined for detection limit in pure bacterial cells and bacteria-spiked plant samples. The assay was applied in monitoring the quantities of the P. cannabina pv. alisalensis DNA over time in inoculated turnip green leaves. As results, the newly developed qPCR assay detected the target DNA in P. cannabina pv. alisalensis suspension as low as 100 CFU/ml and did not detect any of the nontarget bacteria. The qPCR assay detected P. cannabina pv. alisalensis in all the inoculated samples at least 5 days before the symptoms became visible; bacterial quantity increased significantly in the first three days after inoculation but slowed down afterwards. The new qPCR assay for P. cannabina pv. alisalensis detection will facilitate early detection and disease diagnosis, assist research to provide epidemiological insights for the pathogen, and guide implementation of strategies to manage disease and prevent its spread.

Peripheral blood cell count composite score as a prognostic factor in patients with colorectal cancer

Objective: To develop a prognostic prediction model for patients with colorectal cancer based on a peripheral blood cell composite score (PBCS) system. Methods: This retrospective observational study included patients who had primary colorectal cancer without distant metastasis, who did not undergo radiotherapy or chemotherapy before surgery, who did not receive leukocyte or platelet-raising therapy within 1 month before surgery, and whose postoperative pathology confirmed colorectal adenocarcinoma with complete tumor resection. Patients with severe anemia, infection, or hematologic diseases before surgery, as well as those with severe heart, lung, or other important organ diseases or concurrent malignant tumors, were excluded. In total, 1021 patients with colorectal cancer who underwent surgical treatment in the Department of Gastrointestinal Surgery of the Fourth Hospital of Hebei Medical University from April 2018 to April 2020 were retrospectively included as the training set (766 patients) and the internal validation set (255 patients). Additionally, using the same criteria, 215 patients with colorectal cancer who underwent surgical treatment in another treatment group from March 2015 to December 2020 were selected as the external validation set. The "surv_cutpoint" function in R software was used to analyze the optimal cut-off values of neutrophils, lymphocytes, and platelets, and a PBCS system was established based on the optimal cut-off values. The scoring rules of the PBCS system were as follows: Neutrophils and platelets below the optimal cut-off value = 1 point, otherwise 0 points; Lymphocytes above the optimal cut-off value = 1 point, otherwise 0 points. The scores of the three cell types were added together to obtain the PBCS. Univariate and multivariate Cox regression analyses were performed to explore the correlation between patients' clinicopathological features and prognosis, and a nomogram was constructed based on the Cox regression analysis to predict patients' prognosis. The accuracy of the nomogram prediction model was validated using the C-index, calibration curve, and decision curve analysis. Results: The optimal cut-off values for neutrophils, lymphocytes, and platelets were 4.40×109/L, 1.41×109/L, and 355×109/L, respectively. The patients were divided into high and low groups according to the optimal cut-off values of these cells. Survival curve analysis showed that a high lymphocyte count (training set: P=0.042, internal validation: P=0.010, external validation: P=0.029), low neutrophil count (training set: P=0.035, internal validation: P=0.001, external validation: P=0.024), and low platelet count (training set: P=0.041, internal validation: P=0.030, external validation: P=0.024) were associated with prolonged overall survival (OS), with statistically significant differences in all cases. Survival analysis of different PBCS groups showed that patients with a high PBCS had longer OS than those with a low PBCS (P<0.05). Univariate and multivariate Cox regression analysis results showed that aspirin use history, vascular thrombus, neural invasion, CA19-9, N stage, operation time, M stage, and PBCS were independent factors affecting OS (all P<0.05). The PBCS was also an independent factor affecting disease-specific survival (P<0.05), but not progression-free survival (P>0.05). The above independent risk or protective factors were included in R software to construct a nomogram for predicting OS. The C-index (0.873), calibration curve, and decision curve analysis (threshold probability: 0.0%-75.2%) all indicated that the nomogram prediction model had good predictive performance for OS. Conclusion: This study demonstrates that the PBCS constructed based on preoperative peripheral blood levels of neutrophils, lymphocytes, and platelets is an independent factor associated with the prognosis of patients with colorectal cancer. The nomogram model constructed based on this score system exhibits good predictive efficacy for the prognosis of these patients.

Integrated Management of Sclerotinia Stem Rot of Soybean Including Organically-Allowed Fungicides in the Midwest United States

Sclerotinia stem rot (SSR) is an economically important disease of soybean, especially in the Great Lakes region of the United States. Few studies on SSR have considered organic management practices and how integrated management techniques best apply in these systems. Trials in Wisconsin and Indiana aimed to evaluate management techniques available to organic farmers for their efficacy in SSR control while preserving yield. The practices evaluated included genetically resistant cultivars, tillage techniques, and the application of Organic Materials Review Institute (OMRI) listed foliar fungicides. The resistant cultivar had significantly lower SSR than the susceptible cultivar in three site-years (P &lt; 0.01). The resistant cultivar also had significantly greater yield than the susceptible cultivar in two site-years (P &lt; 0.05). Using a roller-crimped rye cover crop resulted in mixed results on SSR and yield. No effect on SSR or yield by OMRI listed foliar fungicides was observed (P &gt; 0.05). Overall, the results emphasize the importance of planting a resistant soybean cultivar to manage SSR in organic farming systems in the Midwestern United States.

Timing of antiviral therapy in patients with hepatitis B virus related hepatocellular carcinoma undergoing hepatectomy.

Globally, hepatocellular carcinoma (HCC) is among the most prevalent and deadly cancers. Hepatitis B virus (HBV) infection is an important etiology and disease progression factor for HCC. Hepatectomy is a widely accepted curative treatment for HCC, but the long-term survival rate is still unsatisfactory due to the high recurrence rate after resection. Preoperative or postoperative antiviral therapy plays an important role in improving the prognosis for HBV-related HCC patients who underwent hepatectomy. However, many patients miss out on the chance to receive long-term preoperative antiviral medication because their HBV and HCC infections are discovered concurrently, necessitating the start of remedial antiviral therapy in the perioperative phase. Therefore, it is of great value to know when antiviral therapy is more appropriate and whether perioperative rescue antiviral therapy can achieve the effect of preoperative long-term antiviral therapy.

Navigating the threat of African swine fever: a comprehensive review.

African swine fever (ASF) is caused by Asfivirus and has become one of the most important diseases of swine in recent years. ASF was an endemic disease of the sub-Saharan Africa but later spread to various parts of the world. The infection in ticks and wild swine, alongside global pork trade, drives its spread and persistence. Once introduced to an area, the disease is difficult to eliminate due to sylvatic, domestic, and tick-swine transmission cycles. Because of the existence of various modes of transmission of the ASF virus, biosecurity measures have not been very successful. The line of treatment is not of much use and the outcome of this disease is usually fatal. The prognosis or the recovery of the animal depends on the virulence of the strain involved. Development of vaccines has been attempted but to date has not been very successful. This review focuses on the basic context of ASF, the challenges associated with it, and the options that might be available to prevent its occurrence which includes the different vaccine development strategies tried and tested till now.

Predicting disease-associated microbes based on similarity fusion and deep learning.

Increasing studies have revealed the critical roles of human microbiome in a wide variety of disorders. Identification of disease-associated microbes might improve our knowledge and understanding of disease pathogenesis and treatment. Computational prediction of microbe-disease associations would provide helpful guidance for further biomedical screening, which has received lots of research interest in bioinformatics. In this study, a deep learning-based computational approach entitled SGJMDA is presented for predicting microbe-disease associations. Specifically, SGJMDA first fuses multiple similarities of microbes and diseases using a nonlinear strategy, and extracts feature information from homogeneous networks composed of the fused similarities via a graph convolution network. Second, a heterogeneous microbe-disease network is built to further capture the structural information of microbes and diseases by employing multi-neighborhood graph convolution network and jumping knowledge network. Finally, potential microbe-disease associations are inferred through computing the linear correlation coefficients of their embeddings. Results from cross-validation experiments show that SGJMDA outperforms 6 state-of-the-art computational methods. Furthermore, we carry out case studies on three important diseases using SGJMDA, in which 19, 20, and 11 predictions out of their top 20 results are successfully checked by the latest databases, respectively. The excellent performance of SGJMDA suggests that it could be a valuable and promising tool for inferring disease-associated microbes.

Identification of Cercospora spp. on corn in North America and baseline flutriafol fungicide sensitivity.

Gray leaf spot (GLS) is an important corn disease reportedly caused by Cercospora zeae-maydis and C. zeina. Recently, flutriafol, a demethylation inhibitor (azole) fungicide received EPA registration as Xyway® LFR®, a product that is applied at planting for management of fungal diseases in corn, including suppression of GLS. In this study, 448 Cercospora spp. isolates were collected in 2020 and 2021 from symptomatic corn leaf samples submitted from the United States and Ontario, Canada. The Cercospora spp. were identified using multi-locus genotyping of the internal transcribe spacer (ITS), elongation factor 1-α (EF1), calmodulin (CAL), histone H3 (HIS), and actin (ACT) gene. Based on the multi-locus phylogenetic analyses, six species were identified; C. cf. flagellaris (n = 77), C. kikuchii (n = 4), C. zeae-maydis (n = 361), Cercospora sp. M (n = 2), Cercospora sp. Q (n = 1), and Cercospora sp. T (n = 3). In subsequent pathogenicity tests using selected isolates from each of these species, only C. zeae-maydis resulted in symptoms on corn with no disease symptoms observed after inoculation with C. cf. flagellaris, C. kikuchii, Cercospora sp. M, Cercospora sp. Q, and Cercospora sp. T. While disease symptoms were observed on soybean following inoculation with C. cf. flagellaris, C. kikuchii, and Cercospora sp. Q, but not the other three species. Fungicide sensitivity of Cercospora spp. to flutriafol was assessed using a subset of 340 isolates. The minimum inhibitory concentration (MIC) to inhibit the growth of Cercospora spp. completely was determined based on growth of each species on flutriafol-amended clarified V8 agar at nine concentrations. The EC50 was also calculated from the same trial by measuring relative growth as compared to the non-amended control. Cercospora zeae-maydis was sensitive to flutriafol with mean MIC values of 2.5 µg/mL and EC50 values ranging from 0.016 to 1.020 µg/mL with a mean of 0.346 µg/mL. Cercospora cf. flagellaris, C. kikuchii, Cercospora sp. M, Cercospora sp. Q, and Cercospora sp. T had mean EC50 values of 1.25 µg/mL, 7.14 µg/mL, 2.48 µg/mL, 1.81 µg/mL, and 2.24 µg/mL respectively. These findings will assist in monitoring the sensitivity to the flutriafol fungicide in Cercospora spp. populations.

Photogallery. HIV infection and neoplastic skin diseases

Malignant neoplasms constitute a group of the most important secondary diseases that develop in people infected with the human immunodeficiency virus (HIV). The first neoplastic disease described in HIV-infected patients is Kaposi's sarcoma. In contrast to other types of Kaposi's sarcoma, its epidemic (AIDS-associated) type is more common in young and middle-aged HIV-positive individuals, especially among men who have same-sex sex. Epidemic Kaposi's sarcoma does not have a favourite localisation. However, there is a tendency to lesions of the scalp (face, ear flaps), upper extremities, anogenital area, oral mucosa. On the trunk, the pathological process spreads along the Langer lines. Skin rashes are represented by spotty, papular, nodular elements of red-brown or purple colour. The aggressive course is characteristic: rapid dissemination and generalisation of rashes. Plasmoblastic lymphoma is associated with immunodeficiency states and is rare. The tumour lesion is usually located in the oral cavity. Few cases of plasmoblastic lymphoma with skin and soft tissue involvement have been described in the literature. Squamous cell skin cancer in the context of HIV infection is a dangerous disease. In people living with HIV, it manifests at a significantly earlier age and is associated with a high risk of local recurrence. The extent of invasion and the likelihood of metastasis are influenced by HIV-associated immunodeficiency. In the vast majority of cases, risk factors and clinical manifestations of basal cell skin cancer do not differ from those in immunocompetent individuals. Basal cell carcinomas rarely metastasise. Nevertheless, metastatic forms lead to unfavourable outcome. Here is a photo gallery of cases of neoplastic skin lesions developed in HIV-positive patients.

New standard area diagram set for assessing black sigatoka in bananas

Black sigatoka, or black leaf streak (BLS), is caused by Pseudocercospora fijiensis and is the most important foliar disease in Musa production. Our goal was to develop a new standard area diagram set (SADs) with high-resolution digital images to aid in visual classification using a quantitative ordinal scale representing six disease severity ranges: 0 (0%), 1 (0 – 5%], 2 (5 – 13%], 3 (13 – 23%], 4 (23 – 40%], 5 (40 – 65%] and 6 (65—100%] and compare visual estimates of BLS severity on banana leaves with those using a previously developed ordinal scale. Three evaluations were conducted by thirteen evaluators and each evaluation was performed at seven-day intervals. The first assessment was done without a SADs. Evaluators subsequently performed two assessments with each the old ordinal scale and the novel SADs. Using the new SADs, evaluators precision, accuracy, repeatability, reproducibility, and residual distributions were improved. The proposed SADs is a tool that helps users in more accurately estimating the severity of BLS.

The challenged urine bicarbonate excretion test in cystic fibrosis: A comprehensive analysis of urine acid/base parameters.

Renal excretion of excess HCO3 - depends on renal cystic fibrosis transmembrane conductance regulator (CFTR) and is impaired in people with cystic fibrosis (pwCF). Urine HCO3 - excretion following oral NaHCO3-loading may be a simple invivo biomarker of CFTR function. In this study, we investigated changes in urine acid/base parameters following oral NaHCO3-loading to comprehensively assess the physiological response to the test and evaluate HCO3 - as the primary test result. Urine acid/base parameters (titratable acid(TA), NH4 +, net acid excretion (NAE) and pH) were measured in bio-banked urine samples from controls (n = 10) and pwCF (n = 50) who completed the challenged urine HCO3 - test. The association between urine acid/base excretion parameters and clinical CF disease characteristics and CFTR modulator therapy-induced changes were assessed. Before treatment, challenged urine acid/base excretion associated with important CF disease characteristics. TA excretion and NAE were lower in pwCF with residual function mutations, 7.9 and 16.6 mmol/3 h, respectively, and lower TA excretion and NAE associated with pancreatic sufficiency. A lower excretion of TA, NH4 +, and NAE associated with a higher percentage of predicted FEV1 (1.3%, 2.5% and 0.8% per mmol/3 h higher, respectively). Modulator treatment decreased TA excretion and NAE (-2.9 and -5.3 mmol/3 h, respectively). Following acute NaHCO3-loading, increased base excretion is mirrored by decreased acid excretion. Urine HCO3 - excretion sufficiently represents the additional urine acid/base parameters as test result. The observed changes in acid excretion support CFTR modulator-induced increase of CFTR-dependent type B intercalated cell HCO3 - secretion and the use of the challenged urine HCO3 - test as a possible CFTR-biomarker.

The sandwich feeding assay for use with first instar nymphs of the Asian citrus psyllid, Diaphorina citri confirms the high susceptibility of this life stage to bacterial pesticidal proteins

Citrus greening or huanglongbing is the most important disease of citrus and threatens citrus production worldwide. As nymphs of Diaphorina citri play a crucial role in the acquisition and transmission of the citrus greening bacterium, suppression of this life stage is particularly important. However, the lack of a tractable feeding assay for use with first instar D. citri nymphs has impeded assessment of the toxicity of bioactives. Of several bacterial pesticidal proteins (BPP) that are toxic to D. citri adults, Mpp51Aa1 and Cry1Ba1, which have LC50 values of 110 and 120 µg/mL respectively in adults, were fed to 1st instar nymphs in a newly developed assay. For this new sandwich feeding assay, parafilm layers containing feeding solution were placed on top of two 35 mm Petri dishes, with a concave surface created on each. Fifty nymphs were transferred to the membrane on one Petri dish, and the second Petri dish placed on the top to create a “sandwich” with the 1st instar nymphs in the middle. Nymphs were fed for four days and the LC50 values for Mpp51Aa1 and Cry1Ba1 were calculated at 6.7 and 41.6 µg/mL respectively. Bioassays with bioengineered plants expressing Cry1Ba1 confirmed that the majority of D. citri mortality occurs during the 1st instar nymph stage, while egg laying adults are much less susceptible. Taken together, these results confirm that 1st instar D. citri nymphs are more susceptible to BPP than adults and demonstrate the utility of the sandwich feeding assay for effective screening of BPPs prior to investment into production of transgenic plants.

Green synthesis of silver nanoparticles for antifungal activity against tomato fusarium wilt caused by Fusariumoxysporum

Fusarium wilt caused by Fusarium oxysporum f. sp. radicis-lycopersici is one of the most important diseases in tomatoes, resulting in severe yield loss and mycotoxin infection in food and feed. The management of Fusarium wilt is based heavily on chemical fungicides, but these chemicals have many environmental concerns. Silver nanoparticles (AgNPs) are gaining importance as emerging resistance-free alternatives of chemical fungicides. In this study, AgNPs were synthesized using the aqueous leaf extract of Viola odorata as a green approach. The antifungal activity of these biosynthesised AgNPs was evaluated in vitro against F. oxysporum using the agar well diffusion method. AgNPs were also evaluated against tomato Fusarium wilt diseases using the foliar spray method in the greenhouse. The characterization of AgNPs revealed that AgNPs were of spherical shape, crystalline in nature, and had an average size of 18 nm. In the antifungal assay, 60 mg/l AgNPs strongly inhibited the growth of F. oxysporum with a growth zone of 18 ± 0.89 mm compared to fungicide (23 ± 1.20 mm) and AgNO3 (52 ± 2.33 mm). Similarly, 60 mg/l of AgNPs when sprayed on tomato plants in the greenhouse showed a survival rate of about 80% with no observable phytotoxic effect. This study provides a first baseline to control Fusarium wilt in tomato using biosynthesised AgNPs without affecting the health of crops.