The impact of combined administration of surfactant and intratracheal budesonide compared to surfactant alone on bronchopulmonary dysplasia (BPD) and mortality rate in preterm infants with respiratory distress syndrome: a single-blind randomized clinical trial.

BACKGROUND: Respiratory distress syndrome (RDS) is one of the most important and common disorders among premature infants. OBJECTIVE: This study aimed to compare the effect of the combination of surfactant and budesonide with surfactant alone on Bronchopulmonary dysplasia (BPD) and mortality rate among premature infants with RDS. METHOD: An outcome assessor-blind randomized clinical trial was conducted on 134 premature infants with RDS who were born in Ayatollah Mousavi Hospital, Zanjan, Iran in 2021. The covariate adaptive randomization method was utilized to allocate participants into two groups (surfactant alone and a combination of surfactant and budesonide). The primary outcomes were BPD and Mortality rate from admission to hospital discharge. The data in this study were analyzed using SPSS software version 18. RESULTS: Overall the comparison of mortality rate and BPD between the two groups did not show a significant difference(p > 0.05). The subgroup results showed that administering surfactant with budesonide to infants under 30 weeks of age significantly reduced the number of deaths compared to using surfactant alone (5 vs. 17). Similar positive effects were observed for the occurrence of Pulmonary Hemorrhage, the need for a second dose of surfactant, oxygen index, mean blood pressure and mean arterial pressure (MAP) in infants under 34 weeks of age compared to more than 34 weeks (p < 0.05). CONCLUSION: These findings suggest that the combination therapy of surfactant and budesonide may be beneficial, particularly in preterm infants with less than 34 weeks gestational age and 1500 birth weight. However, further studies with larger sample sizes and longer follow-up periods are needed to confirm these results and assess long-term outcomes. TRIAL REGISTRATION: The study was registered at the Iranian Registry of Clinical Trials website under the code IRCT20201222049802N1. https://en.irct.ir/user/trial/48117/view . REGISTRATION DATE: 28/02/2021. PUBLIC REPOSITORY: DATA SET: This research data set link is displayed on the Zanjan-Iran Medical Sciences website: https://repository.zums.ac.ir/cgi/users/login? target=https%3 A%2 F/repository.zums.ac.ir/id/eprint .

Wild-type Lactococcus lactis producing bacteriocin-like prophage lysins.

Introduction: Lactococcus is a genus of lactic acid bacteria used in the dairy industry as a starter. Lactococci have been found to produce altogether more than 40 different bacteriocins, ribosomally synthesized antimicrobial proteins. All known Lactococcus spp. bacteriocins belong to classes I and II, which are mainly heat-resistant peptides. No class III bacteriocins, bigger heat-sensitive proteins, including phage tail-like bacteriocins, have been found from the Lactococcus spp. Unlike phage tail-like bacteriocins, prophage lysins have not been regarded as bacteriocins, possibly because phage lysins contribute to autolysis, degrading the host's own cell wall. Methods: Wild-type Lactococcus lactis strain LAC460, isolated from spontaneously fermented idli batter, was examined for its antimicrobial activity. We sequenced the genome, searched phage lysins from the culture supernatant, and created knock-out mutants to find out the source of the antimicrobial activity. Results and discussion: The strain LAC460 was shown to kill other Lactococcus strains with protease- and heat-sensitive lytic activity. Three phage lysins were identified in the culture supernatant. The genes encoding the three lysins were localized in different prophage regions in the chromosome. By knock-out mutants, two of the lysins, namely LysL and LysP, were demonstrated to be responsible for the antimicrobial activity. The strain LAC460 was found to be resistant to the lytic action of its own culture supernatant, and as a consequence, the phage lysins could behave like bacteriocins targeting and killing other closely related bacteria. Hence, similar to phage tail-like bacteriocins, phage lysin-like bacteriocins could be regarded as a novel type of class III bacteriocins.

Multi-Drug Resistance against Second-Line Medication and MicroRNA Plasma Level in Metastatic Breast Cancer Patients.

Background: Circulating microRNAs (miRNAs) can help to predict the chemotherapy response in breast cancer with promising results. The aim of the present study was to investigate the relationships between the miR-199a, miR-663a, and miR-663b expression and chemotherapy response in metastatic breast cancer patients. Methods: This study is a case-control study performed at Yasuj University of Medical Sciences (2018-2021). The expression levels of miR-663a, miR-663b, and miR-199a in the serum of 25 patients with metastatic breast cancer versus 15 healthy individuals were determined by the real-time polymerase chain reaction method. The response to treatment was followed up in a 24-month period. All patients were treated with second-line medications. Two or more combinations of these drugs were used: gemcitabine, Navelbine®, Diphereline®, Xeloda®, letrozole, Aromasin®, and Zolena®. Statistical analyses were performed in SPSS 21.0 and GraphPad Prism 6 software. The expression levels were presented as mean±SD and analyzed by Student's t test. Results: The results and clinicopathological features of patients were analyzed by t test. The statistical analysis showed that miR-663a expression was related to human epidermal growth factor receptor 2 (HER2) status and was significantly lower in the HER2+ than HER2- group (P=0.027). Moreover, the expression of miR-199a and miR-663b was significantly correlated with the response to treatment, in which the expression of miR-199a was higher in the poor-response group (P=0.049), while the higher expression of miR-663b was seen in the good-response group (P=0.009). Conclusion: These findings state that the high plasma level of miR-199a and the low plasma level of miR-663b may be related to chemoresistance in patients with metastatic breast cancer.

Designing a humanized immunotoxin based on DELTA-stichotoxin-Hmg2a toxin: an in silico study.

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.

First Report of Fusarium oxysporum f. sp. vasinfectum causing Fusarium Wilt of Cotton in Kansas, U.S.A.

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 ß-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.

Antifungal Preservation of Food by Lactic Acid Bacteria.

Fungal growth and consequent mycotoxin release in food and feed threatens human health, which might even, in acute cases, lead to death. Control and prevention of foodborne poisoning is a major task of public health that will be faced in the 21st century. Nowadays, consumers increasingly demand healthier and more natural food with minimal use of chemical preservatives, whose negative effects on human health are well known. Biopreservation is among the safest and most reliable methods for inhibiting fungi in food. Lactic acid bacteria (LAB) are of great interest as biological additives in food owing to their Generally Recognized as Safe (GRAS) classification and probiotic properties. LAB produce bioactive compounds such as reuterin, cyclic peptides, fatty acids, etc., with antifungal properties. This review highlights the great potential of LAB as biopreservatives by summarizing various reported antifungal activities/metabolites of LAB against fungal growth into foods. In the end, it provides profound insight into the possibilities and different factors to be considered in the application of LAB in different foods as well as enhancing their efficiency in biodetoxification and biopreservative activities.

Barley-based probiotic food mixture: health effects and future prospects.

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, ß-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 ß-glucans on the growth and features of Lactobacillus strains after fermentation.

Complete Genome Sequence of Cotton Leafroll Dwarf Virus Isolated from Cotton in Texas, USA.

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.

Survival of encapsulated probiotics in pasteurized grape juice and evaluation of their properties during storage.

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<0.05) higher than that in the free cells (8.67 ± 0.12 and 7.57 ± 0.08 log cfu mL-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.

The cell wall compound of Saccharomyces cerevisiae as a novel wall material for encapsulation of probiotics.

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<0.05) enhancement in resistance of L. acidophilus when applying a layer of S. cerevisiae cell wall compound. For MCA and MCYA after 2h exposure to simulated gastric juice, it was revealed a log reduction of 1.53±0.1 and 1.1±0.02 with pH1.55 and in simulated intestinal juice, 2.92±0.04 and 2.42±0.06 with 0.6% bile after previous 1h incubation in gastric conditions, respectively. It can be concluded that the cell wall compound of S. cerevisiae is a suitable protective coating for probiotics and it can improve the survival of probiotics within food products.

Development of a nutraceutical nano-delivery system through emulsification/internal gelation of alginate.

Alginate nano/microspheres are produced by emulsification/internal gelation of sodium alginate dispersed within vegetable oils containing surfactant, followed by CaCl2 addition resulting in hardened particles. In this work, the impact was evaluated of alginate, CaCl2, 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 CaCl2 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, 400ml oil, 0.05M CaCl2 and 100ml surfactant was recognized as the optimized treatment with a reasonable encapsulation efficiency of 5.6% and a nanoparticle size of 785nm.