Antifungal activity of Cowpea (Vigna unguiculata L. Walp) proteins : efficacy, shelf life extension and sensory effects in bread

Natural food preservatives extracted from plants are an emerging market in the food industry. Cowpea Vigna unguiculata L. Walp is a well-established crop around the world, and recent research has revealed that it contains multiple proteins, with potent antifungal and antiviral properties. Exploitation of the antifungal properties of cowpea protein is limited by the lack of characterisation of antifungal proteins, and the lack of knowledge about their efficacy when added to food. Furthermore, the mechanism of antifungal protein activity is not understood fully, and therefore requires further investigation. This study aimed to evaluate the antifungal potency of cowpea seed protein isolate (CPI) against bread yeasts and moulds using microbiological tests, and in a leavened bread application. The research investigated the effects of CPI on shelf life, and the sensory and textural acceptability of bread. Further aims included the characterisation of proteins in CPI with antifungal activity, and establishing an understanding of antifungal activity using computer modelling. The first stage of the study (Chapter 2) involved the preparation of CPI, separation of high and low molecular weight protein fractions by ultrafiltration, and analysis of electrophoretic profiles using SDS PAGE. Analysis of CPI was performed using LCMS/MS, and the proteomics results reported for the first time. Twenty- three proteins with lowest hit number (HN) and highest score were selected from the first ninety-nine hits of matching of LC/MS/MS results to an existing database. The two most abundant proteins (with highest score number and lowest HN) were identified as vicilin (mw 49654), which is a storage protein, and then lipoxygenase (mw 97284) which is a metabolism protein. The second stage of the study (Chapter 3) involved testing CPI activity against fungal growth using a micro spectrophotometric assay (micro plate method) and the spread plate method. Activity was tested against known bread spoilage moulds: Penicillium chrysogenum, Penicillium brevicompactum, Penicillium hirsutum, Aspergillus versicolor and Eurotium rubrum. The results showed CPI exhibited high antifungal activity against P. chrysogenum, P. brevicornpactum, P hirsutum and E. rubrum; no statistically significant effect was seen against A. versicolor. CPI exhibited different ranges of inhibition towards the same species at different concentrations demonstrating that the antifungal effect was concentration dependent. The antifungal activity of CPI was unaffected by heat treatment or protease treatment, indicating the antifungal components are heat stable and protease resistant. The antifungal activity of proteins in the CPI ultrafiltrate (10 kDa molecular weight cut off point) was increased by comparison with CPI and CPI retentate. The third stage of the study (Chapter 4) involved testing the effect of CPI addition on shelf life, and the sensory and textural properties of leavened bread. A concentration of 2.3% of CPI showed the best resistance to fungal growth during the storage period. No growth was observed throughout the 8 day storage period at room temperature, whereas control samples began to show contamination on the fourth day of storage. The CPI filtrate (2.3%) showed better shelf life extension than CPI after 25 days, confirming the antifungal activity of low molecular weight proteins. Inclusion of 2.3% CPI did not significantly affect the sensory or textural acceptability of the bread. Although the hardness of bread containing CPI increased significantly after 3 days compared to the control, sensory acceptability was within acceptable limits. The fourth stage of the study (Chapter 5) involved understanding the mechanism of action of defensins at the fungal cell wall. The hypothesis for defensin as a major antifungal protein in CPI was based on the results of the antifungal activity of the low molecular weight fraction of CPI, the heat stable and protease resistant antifungal properties of CPI, and confirmation of the presence of defensins in the proteomics results. The computer model detailing the simulation of defensin activity showed the adsorption of defensin molecules to the surface of a phospholipid bilayer membrane leading to a disordering of the membrane that would ultimately lead to disruption of cell metabolism

The New Dagum-X Family of Distributions: Properties and Applications

Various statistical distributions are still being used extensively over the previous decades for modeling data in numerous areas such as engineering, sciences, and finance. Nonetheless, in a lot of applied areas, there is a continuous need for expanded forms of these distributions. However, many common distributions do not fit the data well. Thus, new distributions have been constructed in literature. The purpose of this article is to present a new family of distributions using the Dagum distribution as a generator and to study its properties such as hazard rate functions, moments, quantile function, ordered statistics and Renyi entropy. Moreover, one sub model called Dagum-Frechet distribution is discussed with some of its properties. The maximum likelihood estimation is employed to estimate the parameters of the proposed distribution, and the confidence intervals are obtained. Finally, two real data sets are analyzed to illustrate the performance of the purposed distribution

Antifungal Activity of Syzygium aromaticum (Dianthus) against toxigenic Rhizopus stolonifera and its immunomodulatory effects in aflatoxin-fed mice

Purpose: To evaluate the antifungal potency of Syzygium aromaticum (Dianthus) seed extract against the growth and aflatoxin production of Rhizopus stolonifer, and its immunomodulatory effect.Methods: Disc diffusion method was used for assay of antifungal effect of aqueous extract of Dianthus. Lymphoid cell counts, total and differential peritoneal exudate cell counts (PEC), phagocytic activity of PEC, and plaque-forming activities were determined. In addition, E-rosette-forming cells (RFC), T-cell mitogenesis cells and liver functions were measured.Results: The aqueous extract of Dianthus (50 %) exhibited high inhibition zone against most isolates of R. stolonifera. It produced significant increases in the number of splenocytes, as well as in the absolute number and relative proportion of macrophages (p < 0.05). The extract also produced a gradual increase in the scavenging activity of PEC, and significant reduction in serum ALT, relative to control.Conclusion: These results suggest that Dianthus modifies biological responses by enhancement of the immune system, activation of phagocytosis, boosting of immune response, and prevention of liver damage.Keywords: Aflatoxin, Syzygium aromaticum, Rhizopus stolonifer, Lymphocytes, T-cells, B-cells, Macrophage

Investigating Cardiovascular Activation of Young Adults in Routine Driving

We report on a naturalistic study investigating the effects of routine driving on cardiovascular activation. We recruited 21 healthy young adults from a broad geographic area in the Southwestern United States. Using the participants' own smartphones and smartwatches, we monitored for a week both their driving and non-driving activities. Monitoring included the continuous recording of a) heart rate throughout the day, b) hand motion during driving as a proxy of persistent texting, and c) contextualized driving data, complete with traffic and weather information. These high temporal resolution variables were complemented with the drivers' biographic and psychometric profiles. Our analysis suggests that anxiety predisposition and high speeds are associated with significant cardiovascular activation on drivers, likely linked to sympathetic arousal. Surprisingly, these associations hold true under good weather, normal traffic, and with experienced drivers behind the wheel. The said findings call for attention to insidious effects of apparently benign drives even for people in their prime. Accordingly, our research contributes to intriguing new discourses on driving affect and personal health informatics

Aged garlic extract potentiates doxorubicin cytotoxicity in human breast cancer cells

Purpose: To investigate the potential chemo-sensitizing effect of aged garlic extract (AGE) on doxorubicin (DOX) in breast cancer cells (MCF-7), and the possible underlying mechanisms.Methods: Human breast cancer cell line (MCF-7) was treated with AGE and DOX. The cytotoxic effects of AGE and DOX were investigated via cell cycle analysis and apoptosis induction, using flow cytometry. Mechanistic studies involved the determination of cellular uptake of DOX and  p-glycoprotein (P-gp) activity.Results: Combined treatment of MCF7 cells with AGE and DOX produced no significant effect at AGE dose of 10 mg/mL. However, co-treatment with AGE at doses of 50 and 93 mg/mL enhanced the cytotoxicity of DOX on MCF-7 cells, with IC50 values of 0.962 and 0.999 μM, respectively, whencompared with 1.85 μM DOX alone. Moreover, Annexin V-FITC and PI techniques showed that AGE significantly increased percentage of cells in late apoptosis. Besides, AGE-DOX treatment significantly increased cellular uptake of DOX and inhibited P-gp activity, when compared with DOX alone (p < 0.05).Conclusion: AGE enhances the cytotoxic effect of DOX on MCF-7 cells, most likely due to cell cycle distribution, stimulation of apoptosis, increased uptake of DOX by MCF7, and inhibition of P-gp activity. Keywords: Aged garlic extract, Doxorubicin, Breast cancer, MCF-7 cell line, P-glycoprotein, Apoptosis, Cell cycl

Electroanalysis of benzalkonium chloride in ophthalmic formulation by boron-doped diamond electrode

Benzalkonium chloride (BAK) is a mixture of alkyl benzyl dimethyl ammonium chlorides, which is used primarily as a biocide, surfactant, preservative, and antimicrobial agent in the pharmaceutical industry, in particular in ophthalmologic and nasal solutions. However, BAK may cause harmful consequences on the eye structures of the anterior segment. Control of BAK identity and content is necessary by applying a sensitive detection method. This study unravels the use of a glassy carbon (GC) electrode and a pristine boron-doped diamond electrode (BDD) for the detection of four BAK homologs in a non-aqueous medium using square wave voltammetry (SWV). The BDD provided more reproducibility of the oxidation potential than GC with a correlation coefficient of 0.999. The irreversible oxidation peak was very broad and deconvoluted into 3 peaks corresponding to C12, C14, and combined C16–C18 to reflect their concentration ratio in the mixture. The method was then extended to the detection of the C12 homolog in the ophthalmic formulations with a limit of detection (LOD) of 0.4 μg/mL. The estimated BAK levels in three ophthalmic formulations were in agreement with the specified values by the manufacturers. The results were validated by high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection, confirming the presence of a single homolog (C12) in the eye drops

The socioeconomic burden of spinal muscular atrophy in Saudi Arabia: a cross-sectional pilot study

BackgroundSpinal muscular atrophy (SMA) is a rare debilitating condition with a significant burden for patients and society. However, little is known about how it affects Saudi Arabia's population. The socioeconomic and medical characteristics of affected SMA patients and their caregivers are lacking.PurposeThis study aimed to describe the socioeconomic and medical characteristics of SMA patients and caregivers in Saudi Arabia.Patients and methodsA cross-sectional questionnaire-based study was conducted using snowball sampling. Assessment tools including EuroQol (EQ-5D-5L) and visual analog scale (EQ-VAS), Generalized Anxiety Disorder 7-item (GAD-7), Patient Health Questionnaire (PHQ-9), and Costs for Patients Questionnaire (CoPaQ) were used to assess the quality of life (QoL), anxiety, depression, and out-of-pocket expenditures.ResultsSixty-four caregivers of SMA patients participated. Type I patients had higher sibling concordance, ICU hospitalization, and mechanical support needs. Type III patients had better QoL. Type I patients' caregivers had higher depression scores. Type III patients' caregivers had higher out-of-pocket expenditures. Forty-eight percent received supportive care, while others received SMA approved therapies.ConclusionSMA imposes a significant socioeconomic burden on patients and caregivers, requiring more attention from the healthcare system. Access to innovative therapies varied across SMA types. Pre-marital screening and early detection are crucial to reduce disease incidence and ensure timely treatment

Clinical Correlation Between Back Pain and Osteoarthritis Among Adults: A Cross-Sectional Study

Objective: To investigate the clinical correlation between back pain and the role of osteoarthritis in the etiology of this pain. Methods: This research employs a cross-sectional study design to investigate the clinical correlation between back pain and osteoarthritis in adults. Cross-sectional studies provide a snapshot of data at a specific time, making them suitable for assessing the prevalence and potential associations between variables of interest. Results: The study included 1106 participants. The most frequent age among them was 40-50 years (n= 486, 43.9%), followed by 18-28 years (n= 340, 30.7%). The most frequent gender among study participants was female (n= 560, 50.6%) followed by male (n= 546, 49.4%). The most frequent marital status among study participants was married (n= 702, 63.5%), followed by single (n= 353, 31.9%). The most frequent job among study participants was a government job (n= 480, 43.4%), followed by a private sector job (n= 177, 16%). Duration of suffering from back pain among study participants with most of them being 1 to 5 years (n= 505, 45.7%), followed by nothing (n= 373, 33.7%), and the least was 23 years and above (n= 13, 1.2%). Participants were asked about evaluating the impact of back pain on life. For personal care, the most frequent was (currently, I do not feel any pain) (n= 514, 46.5%). The severity of pain, the most frequent, was (I can usually take care of myself and do my own things without adding to my pain) (n= 923, 83.5%). The lift and move things, the most frequent was (I can lift heavy objects without it adding to my pain) (n= 524, 47.4%). The walking, the most frequent was (The pain does not prevent me from walking any distance) (n= 801, 72.4%). The Sitting, the most frequent was (I can sit in any chair for as long as I want) (n= 661, 59.8%). The stand-up, most frequent was (I can stay standing as long as I want without increasing my pain) (n= 523, 47.3%). For sleep, the most frequent answer was (My sleep is never disturbed by pain) (n= 611, 55.2%). The Social-life, most frequent was (My social life is normal and does not cause me more pain) (n= 860, 77.8%). Conclusion: The results of the study showed that most of the participants were married and taught in government jobs. The majority of them do not suffer from severe back pain that affects their lives. In addition, most of the study participants had good and effective communicatio

An Evaluation of Psychological Well-Being among Physicians and Nurses in Makkah’s Major Hospitals

Background: Physicians and nurses experience poor psychological well-being relative to other employees in healthcare fields. This study aimed to evaluate the psychological well-being among physicians and nurses in Makkah’s major hospitals. Methods: In this cross-sectional study, 460 physicians and nurses from seven major hospitals in Makkah were recruited to investigate their psychological well-being using the General Health Questionnaire-12 (GHQ-12) based on social dysfunction, anxiety, and confidence loss. Results: Over half (64.3%) of the physicians and nurses in this study scored at or above the GHQ-12 cut-off point (12), which is a positive result for poor psychological well-being. There were significant differences in the psychological well-being mean between Saudis and non-Saudis (t = 2.203, p = 0.028), years of work experience (t = 3.349, p = 0.001), hospitals (F = 2.848, p = 0.010), attending psychological support sessions (t = 2.082, p = 0.038), and history of visiting psychological clinics (t = −4.949, p < 0.001). There was also a significant association between the three GHQ-12 factors and the participants’ socio-demographic characteristics. Conclusion: The psychological well-being of physicians and nurses is low. The alarming number of physicians and nurses suffering from social dysfunction, anxiety, and loss of confidence should be addressed in Makkah’s major hospitals. The employee assistance program (EAP) could be highly valuable and effective for addressing the well-being of employees and their personal problems that may impact their work performance, conduct, health, and overall well-being at the Ministry of Health