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Fusarium wilt is one of the most devastating diseases of cotton (Gossypium spp.). It is caused by the soil-borne pathogen Fusarium oxysporum Schlechtend. f. sp. vasinfectum (Atk.) Snyd.Hans (Atkinson, 1892). To date, eight races of F. oxysporum f. sp. vasinfectum have been reported worldwide based on their diverse genetics and reactions to the host (Cianchetta, et al. 2015). Fusarium wilt symptoms appear at all developing stages. The fungus enters through the roots and colonizes the vascular system, leading to discoloration and wilting (Davis et al. 2006). During a survey in the July 2021 growing season, cotton plants showing typical wilting symptoms, stem discoloration, and root rot (Fig. 1A and B) were observed in two cotton fields in Sumner County, Kansas. In order to confirm the causal agent, root tissues and lower parts of the stems were collected from 25 diseased cotton plants and cleaned as previously described by Larren et al, (2001). Small segments (1-2 cm) of roots and stems were incubated on potato dextrose agar (PDA) at room temperature. The isolates were purified with the single spore method and 23 of 25 isolates (92%) showed typical morphology of F. oxysporum, with a white-peach pigmentation as reported previously (Leslie and Summerell 2008). Two isolates, CK13B and CK18A, one from each of the two fields were selected, and genomic DNA was extracted with E.Z.N.A Fungal DNA Mini kit (Omega Bio-tek, Norcross, GA). Polymerase chain reaction (PCR) assays were performed as reported previously (Ortiz, et al. 2017) to amplify and sequence a portion of three nuclear genes: translation elongation factor (EF-1α), phosphate: H* symporter (PHO), and β-tubulin (BT). BLASTn analysis of CK13B sequences showed 93-98% identity with F. oxysporum f. sp. vasinfectum isolates originating from Australia, with 93.43% (1706/1826 pb) identity of EF-1α to isolate AuSeed14 (KT323873), 95.11% (1849/1944 bp) of PHO to isolate AuK24232 (KT323909), and 97.56% (1840/1886 bp) identity of BT to isolate AusSeed14 (KT323833). Sequences of CK13B for EF1-α, PHO, and BT genes were submitted to GenBank, accession no. of ON754247, ON754248, and ON754249, respectively. The CK18A isolate showed high identity with F. oxysporum f. sp. vasinfectum races 1, 2, and 6 lineage isolates (Ortiz et al., 2017), with 99.89% (1822/1824 bp) identity of the EF1-α gene to isolate CDR238 (KT323838) from Arkansas, 98.36% (1916/1948 bp) identity of the PHO gene to isolate CDR1131 (KT323887) from Louisiana and 98.78% (1864/1887 bp) identity of the BT gene to isolate ATCC36198 (KT323799) from Brazil. Sequences of the CK18A isolate were submitted to GenBank, accession no. ON651444, ON725043, and ON725044, respectively. Pathogenicity of both isolates was tested on G. hirsutum as previously described (Kim et al. 2005) with three replicates per isolate and a control treatment in a growth chamber. Briefly, cotton seedlings were maintained with 12 h light/dark cycle at 25-20 °C. At the first true leaf stage seedlings were uprooted, roots were cleaned by rinsing with sterile water then placed in a conidial suspension (1×106 conidia/ml) for 2 h while the healthy control seedlings were dipped in water only. Plants were put into sterile soil, and the results were observed and recorded every seven days for a period of 30 days. The inoculated plants showed the same symptoms as those observed in the fields, wilting, leaf chlorosis, and necrosis. Both isolates were re-isolated from the roots of the inoculated plants. DNA was extracted and used for PCR with the specific gene primers as listed above. The sequences matched those of the original isolates, completing the Koch's pustulates. To our knowledge, this is the first report of F. oxysporum f. sp. vasinfectum causing Fusarium wilt disease of cotton in Kansas. These results will help growers select cultivars and design disease control strategies accordingly (Davis, et al., 2006). Further work is needed to determine the specific races in the area.

Breast cancer remains the most frequently diagnosed cancer and the principal cause of mortality by malignancy in women. HER2 positive subtype includes 15-20% of breast cancer cases. This receptor could be an appropriate mark for targeting breast cancer cells. Immunotherapy methods compared to current cancer treatment methods have the lowest side effects. DELTA-stichotoxin-Hmg2a is isolated from the sea anemone and kills cells through pore formation. In the current study, we designed and evaluated an immunotoxin composed of pertuzumab and DELTA-stichotoxin-Hmg2a-derived scFv by bioinformatics tools. The designed immunotoxin was constructed using the amino acid sequences. Then, secondary structure and physico-chemical features were studied, and the tertiary structure of the immunotoxin was built according to the homology modeling methods. The validation and allergenicity of the model were assessed. The immunotoxin and receptor were docked and molecular dynamics simulation indicated the construct stability. The analysis results indicated that the construct is a stable protein that could have a natural-like structure and would not be an allergen, so this immunotoxin could effectively target HER2 receptors. Therefore, our designed immunotoxin could be an appropriate immunotoxin against HER2-positive breast cancer and could be a challenging topic for future in vitro and in vivo studies.

The authors have requested that this preprint be removed from Research Square.

Consumers around the globe are increasingly aware of the relation between nutrition and health. In this sense, food products that can improve gastrointestinal health such as probiotics, prebiotics and synbiotics are the most important segment within functional foods. Cereals are the potential substrates for probiotic products as they contain nutrients easily assimilated by probiotics and serve as the transporters of Lactobacilli through the severe conditions of gastrointestinal tract. Barley is one of the important substrates for the probiotic formulation because of its high phenolic compounds, β-glucans and tocols. The purpose of this review is to examine recent information regarding barley-based probiotic foods with a specific focus on the potential benefits of barley as a substrate for probiotic microorganisms in the development of dairy and nondairy based food products, and to study the effects of food matrices containing barley β-glucans on the growth and features of Lactobacillus strains after fermentation.

Cotton leafroll dwarf virus (CLRDV; genus, Polerovirus; family, Luteoviridae) was first described in Alabama. In this study, we present the complete genome (5,865 nucleotides) sequence of a CLRDV isolate (CS4) that was collected from cotton during the 2019 growing season in Texas.

Yeast cell wall is known as a food grade ingredient which is recently being used increasingly as a novel coating for encapsulation of different materials in the food industry. This application is limited to core materials smaller than yeast in size. In this study, we have tried to encapsulate larger particles by crushing yeast cells. Hence, probiotic bacteria of Lactobacillus acidophilus and Bifidobacterium bifidum were encapsulated firstly by calcium alginate using the emulsion method and these microbeads were coated again by Saccharomyces cerevisiae cell wall compound and another layer of calcium alginate. The average diameter of microcapsules for single layer microbeads (M), microbeads coated by two layers of alginate (MCA), and microbeads coated by a layer of yeast cell and two layers of alginate (MCYA) were 54.25±0.18, 77.43±8.24 and 103.66±13.33μm, respectively. In simulated gastrointestinal conditions, there was a significant (P

Summary In this study the feasibility of incorporation of probiotic microcapsules coated with fragmented yeast cell wall in grape juice was evaluated during 60 days at 4 °C. Lactobacillus acidophilus and Bifidobacterium bifidum were encapsulated in alginate microbeads and coated with fragmented Saccharomyces cerevisiae cell wall and calcium alginate and were added into grape juice. At the end of storage, the survival of probiotics was higher than recommended minimum value (107 cfu mL−1) and the results demonstrated that applying yeast cell wall layer for L. acidophilus microcapsules significantly enhanced its survival while did not affect the survival of B. bifidum (P > 0.05). Generally, probiotic grape juice showed decrease in °Brix, pH and colour and increase in acidity and turbidity during storage and the presence of yeast wall layer had no significant effect on its properties expect colour and turbidity. Overall acceptance of grape juices containing yeast cell wall-coated microcapsules scored the least.

In the United States, cotton leafroll dwarf disease, caused by cotton leafroll dwarf virus (CLRDV, Polerovirus, Luteoviridae), was first reported from cotton (Gossypium hirsutum L.) fields in Alabama (Avelar et al. 2019) and later Mississippi (Aboughanem-Sabanadzovic et al. 2019) and Georgia (Tabassum et al. 2019). During a survey of cotton fields in Barber County, Kansas, in the 2019 growing season, cotton plants showed virus-like symptoms including leaf yellowing, discoloration, reddening, and short internodes with reduced or small boll sets. Disease incidence based on virus-like symptoms was ∼1 to 15% depending on the affected fields. Eleven symptomatic and seven apparently asymptomatic leaf samples from 18 independent cotton plants (cv. PhytoGen 210) were collected in commercial cotton fields and tested for the presence of the virus in the Plant Virology Lab at the University of Tulsa, Oklahoma. Total RNA was extracted from all 18 samples by the Spectrum Plant Total RNA Kit (Sigma-Aldrich) and was synthesized into complementary DNA (cDNA) using the Superscript IV Reverse Transcription Kit (ThermoFisher Scientific) according to the manufacturer’s instructions. The polymerase chain reaction (PCR) was performed with primers CLRDV3675F and Pol3982R as described previously (Sharman et al. 2015) to amplify a specific PCR product of 310 bp of the ORF3-5 fragment of the CLRDV. cDNA of CLRDV for the specific fragment was obtained from the Plant Disease Diagnostic Lab at Auburn University and was used as a positive control. A PCR product of the predicted size (310 bp) was obtained from seven symptomatic cotton samples and the positive control, whereas no bands were present in four symptomatic and seven asymptomatic samples, as well as the PCR negative control. RT-PCR was repeated twice with these samples and obtained the same results as above. The PCR products of all positive samples were directly sequenced, and sequence comparison showed 97 to 99% identity with the CLRDV-Alabama isolate (MN071395). To further confirm the presence of CLRDV in these samples, two additional pair of primers—AL674F and AL1407R for partial P0/P1 (Avelar et al. 2019) and RdRpF2 and RdRpR1 for partial RdRp (Aboughanem-Sabanadzovic et al. 2019)—were used to amplify the 694- and 770-bp product of the CLRDV genome, respectively. The expected PCR bands were obtained from all seven symptomatic cotton plants but none from asymptomatic samples. PCR products of 694 and 770 bp from two samples (KS-7 and KS-10 isolates) were purified, cloned into pGEM-T Easy vector (Promega), and transformed into Escherichia coli DH5α cells (New England Bio Labs). At least three clones were sequenced using the Applied Biosystem 3130 Genetic Analyzer at the University of Tulsa. Sequences obtained in this study for partial P0/P1and RdRP genes of KS-7 and KS-10 isolates were deposited in GenBank (MN809925 to 28). Nucleotide sequence comparisons and BLASTn analysis of partial P1/P0 (MN809927 to 28) showed 98 to 99% identity with corresponding sequences of Alabama isolates (MH883237 and MN071395) and 92 to 93% identity with typical and 91 to 92% identity with atypical CLRDV isolates from Argentina and Brazil. Similarly, partial RdRP genes (MN809925 to 26) showed 94 to 98% identity with the corresponding sequences of available CLRDV isolates from Mississippi (MK512759, MK512762 to 64) and Alabama (MN071395) and 93 to 96% identity with typical and 92 to 95% identity with atypical CLRDV isolates from Argentina and Brazil. This is the first report of CLRDV infection and occurrence in cotton fields in Kansas. Cotton acreage in Kansas has increased since 2018 by 40% (NASS 2018), and the presence of CLRDV could be a detrimental disease for cotton production. Therefore, further studies are needed, including other cotton-growing counties in Kansas, to determine the distribution, epidemiology, vector transmission, and population structure of this virus, which would be helpful in the management of CLRDV.

In this study, probiotic bacteria such as Lactobacillus acidophilus and Bifidobacterium bifidum were encapsulated in alginate beads with a mean diameter of 54.25 ± 0.18 µm by internal gelation. Encapsulated and free cells as control samples were then added to the pasteurized grape juice and stored for 60 days. At the end of the storage period, the survivability of the bacteria in the encapsulated samples was significantly ( P−1 for L. acidophilus and 8.27 ± 0.05 and 7.53 ± 0.07 log cfu mL−1 for B. bifidum for encapsulated and free forms, respectively). The results generally showed a decrease in °Brix, pH, and color, whereas acidity and turbidity have increased the in probiotic grape juice after the storage period of 60 days. For all treatments, the numbers of surviving cells were more than the recommended minimum (107 cfu g−1) at the end of the storage period.

Farsesh barite in the central part of Iranian Sanandaj-Sirjan zone is a sample of epigenetic hydrothermal mineralization in dolomitized limestone, which provides appropriate chemicophysical conditions making the passage of mineral-bearing fluids possible. Barite veins may range from a few centimeters to 2 m in thickness that increases downward. The microthermometry measurements obtained from more than 30 fluid inclusions show relative homogenization temperatures ranging from 125 to 200 °C with an average of 110 °C for Farsesh barite deposits. The mean salinity measured proves 16 times as much as weight percentage of NaCl for barite. Coexistence of liquid- and vapor-rich fluid inclusions in barite minerals may provide an evidence of boiling in ore veins. Moreover, occurrence of bladed calcite, high-grade ore zones, and presence of hydrothermal breccia are all consistent with boiling. Thermometric studies indicate that homogenization temperatures (Th) for primary and pseudosecondary fluid inclusions in barite range from 125 to 200 °C with an average of 1,100 °C. The δ34S values of barite also lie between 8.88 and 16.6 %. The relatively narrow spread in δ34S values may suggest uniform environmental conditions throughout the mineralization field. Thus, δ34S values are lower than those of contemporaneous seawater, which indicates a contribution of magmatic sulfur to the ore-forming solution. Barite is marked by total amounts of rare Earth elements (REEs) (6.25–17.39 ppm). Moreover, chondrite-normalized REE patterns of barite indicate a fractionation of light REEs (i.e., LREEs) from La to Sm, similar to those for barite from different origins. The LaCN/LuCN ratios and chondrite-normalized REE patterns reveal that barite in Farsesh deposit is enriched in LREEs compared with heavy rare Earth elements (HREEs). Similarity between Ce/La ratios in barite samples and those found in deep-sea barite supports its marine origin. Lanthanum and Gd exhibit positive anomalies, which are common features of chemical marine sediments. Cerium shows a negative anomaly in most samples inherited from the negative Ce anomaly of hydrothermal fluid that is mixed with seawater at barite precipitation. The available data including tectonic setting, host rock characteristics, REE geochemistry, and sulfur isotopic compositions may support a hydrothermal submarine origin for Farsesh barite deposit.

Alginate nano/microspheres are produced by emulsification/internal gelation of sodium alginate dispersed within vegetable oils containing surfactant, followed by CaCl 2 addition resulting in hardened particles. In this work, the impact was evaluated of alginate, CaCl 2 , oil and surfactant content on the size and encapsulation efficiency of nanocarriers containing peppermint phenolic extract and prepared by a low energy internal gelation technique. The results revealed that size of nanoparticles decreased at higher oil and surfactant contents, higher molarity of CaCl 2 and lower alginate concentrations. Also, it was found that the encapsulation efficiency was inversely proportional to the size of nanoparticles, and the impact of alginate concentration and surfactant content was markedly higher than the other two factors. The composition of 0.5% alginate, 400 ml oil, 0.05 M CaCl 2 and 100 ml surfactant was recognized as the optimized treatment with a reasonable encapsulation efficiency of 5.6% and a nanoparticle size of 785 nm.