HeLa cell segmentation using digital image processing.

Computational cell segmentation is a vital area of research, particularly in the analysis of images of cancer cells. The use of cell lines, such as the widely utilized HeLa cell line, is crucial for studying cancer. While deep learning algorithms have been commonly employed for cell segmentation, their resource and data requirements can be impractical for many laboratories. In contrast, image processing algorithms provide a promising alternative due to their effectiveness and minimal resource demands. This article presents the development of an algorithm utilizing digital image processing to segment the nucleus and shape of HeLa cells. The research aims to segment the cell shape in the image center and accurately identify the nucleus. The study uses and processes 300 images obtained from Serial Block-Face Scanning Electron Microscopy (SBF-SEM). For cell segmentation, the morphological operation of erosion was used to separate the cells, and through distance calculation, the cell located at the center of the image was selected. Subsequently, the eroded shape was employed to restore the original cell shape. The nucleus segmentation uses parameters such as distances and sizes, along with the implementation of verification stages to ensure accurate detection. The accuracy of the algorithm is demonstrated by comparing it with another algorithm meeting the same conditions, using four segmentation similarity metrics. The evaluation results rank the proposed algorithm as the superior choice, highlighting significant outcomes. The algorithm developed represents a crucial initial step towards more accurate disease analysis. In addition, it enables the measurement of shapes and the identification of morphological alterations, damages, and changes in organelles within the cell, which can be vital for diagnostic purposes.

Changes in phenolics and flavonoids in amaranth and soybean sprouts after UV-C treatment.

Sprouts, mainly from cereals, legumes, and some pseudo-cereals, are rich in nutrients and contain biocompounds, making them attractive for consumption. This research study aimed to develop treatments with UV-C light in soybean and amaranth sprouts and evaluate their effect on biocompounds content, compared with chlorine treatments. UV-C treatments were applied at distances of 3 and 5 cm and times of 2.5, 5, 10, 15, 20, and 30 min, whereas chlorine treatments were applied as immersion in solutions at 100 and 200 ppm for 15 min. Phenolics and flavonoid content were higher in UV-C-treated sprouts than in those treated with chlorine solutions. Ten biocompounds were identified in soybean sprouts, with increasing in apigenin C-glucoside-rhamnoside (105%), apigenin 7-O-glucosylglucoside (237%), and apigenin C-glucoside malonylated (70%) due to UV-C application (3 cm, 15 min); for amaranth sprouts, five biocompounds were identified, with higher contents of p-coumaroylquinic acid (17.7%) after UV-C treatment (3 cm, 15 min). The best treatment to achieve the highest bioactive compounds concentration was UV-C at a distance of 3 cm for 15 min, without significant modification on the color parameters, Hue and chroma. PRACTICAL APPLICATION: UV-C can be used to increase the biocompound content in amaranth and soybean sprouts. Nowadays, there is UV-C equipment available for industrial applications. In this way, sprouts may be maintained as fresh through this physical technology, and they will retain or increase the concentration of health-related compounds.

Daily Intake of a Phaseolus vulgaris L. Snack Bar Attenuates Hypertriglyceridemia and Improves Lipid Metabolism-Associated Plasma Proteins in Mexican Women: A Randomized Clinical Trial.

Current efforts to prevent dyslipidemia are focused on the development of functional products as an alternative for hypertriglyceridemia management. This study assessed the metabolic effect of the daily consumption of a bean and oats snack bar (BOSB) on hypertriglyceridemia biomarkers among Mexican women. An 8-weeks randomized parallel clinical trial (ID: NCT0496694, https://clinicaltrials.gov/ct2/show/NCT04966494) was conducted with 26 hypertriglyceridemic women allocated to BOSB group (TG = 208.18 ± 56.97 mg/dL) and control group (TG = 182.28 ± 51.39 mg/dL). Only the BOSB group consumed 50 g of the product per day. Fasting blood samples were taken from women with an adherence ≥ 90%. A targeted proteomic analysis with plasma samples of control and BOSB groups were conducted using a human obesity antibody array kit and bioinformatic tools provided by the Ingenuity Pathways Analysis (IPA) software. Serum TG levels in the BOSB group decreased by 37.80% (132.04 ± 27.83 mg/dL) compared with the control group (178.87 ± 32.01 mg/dL); glucose levels decreased by 5.69% in the BOSB group (87.55 ± 3.36 mg/dL). A modest body weight (5%) reduction was also found. Forty proteins were differentially modulated by the BOSB consumption (fold change > 1.2). The proteomic analysis revealed the involvement of BOSB bioactives in prevention of monocytes recruitment and localized inflammatory response, inhibition of pre-adipocyte maturation and adipogenesis, inhibition of hepatic b-oxidation, and potential satiety regulation. These results are promising since the mere intervention with the BOSB reduced serum TG without diet restriction, giving insights for further research in prevention of hypertriglyceridemia.

Analysis of virulence traits of Staphylococcus aureus isolated from bovine mastitis in semi-intensive and family dairy farms.

BACKGROUND: Staphylococcus aureus is one of the main microorganisms that causes bovine mastitis, and its well-known virulence characteristics and interactions with the environment are used to aid the design of more efficient therapies. OBJECTIVES: To determine whether the virulence traits, such as antibiotic resistance and biofilm-forming and internalization abilities, of S. aureus isolated from bovine mastitis are related to dairy production system types. METHODS: The study was performed in the Mexican states of Guanajuato and Michoacan. Semi-intensive dairy farms (SIDFs) and family dairy farms (FDFs) (454 and 363 cows, respectively) were included. The 194 milk samples from mastitis affected quarters were collected and 92 strains of S. aureus were isolated and identified by biochemical and molecular tests. Antibiotic resistance, biofilm and internalization assays were performed on 30 randomly selected isolated strains to determine virulence traits, and these strains were equally allocated to the 2 dairy production systems. RESULTS: All 30 selected strains displayed a high degree of resistance (50%-91.7%) to the antibiotics tested, but no significant difference was found between SIDF and FDF isolates. S. aureus strains from SIDFs had an average biofilm forming capacity of up to 36% (18.9%-53.1%), while S. aureus strains from FDFs registered an average of up to 53% (31.5%-77.8%) (p > 0.05). Internalization assays revealed a higher frequency of internalization capacity for strains isolated from FDFs (33.3%) than for those isolated from SIDFs (6.7%) (p > 0.05). fnbpA gen was detected in 46.6% of FDF strains and 33.3% of SIDF strains, and this difference was significant (p < 0.05). CONCLUSIONS: Our findings show that the virulence traits of S. aureus isolates analyzed in this study, depend significantly on several factors, such as phenotype, genotype, and environmental conditions, which are significantly related to dairy production system type and daily management practices.

Antibiotic susceptibility of Staphylococcus aureus isolated from subclinical bovine mastitis cases and in vitro efficacy of bacteriophage.

Staphylococcus aureus is an opportunistic pathogen that may cause severe infections in livestock, and represents the major cause of mastitis in dairy cows. Currently, instead of using antibiotics, new strategies are sought to reduce this clinical health problem. The aim of this study was to determine the efficacy of phage therapy to kill S. aureus strains obtained from farms located at the State of Guanajuato, México. Thirty-six S. aureus strains from cow milk with subclinical mastitis were isolated and identified, and the susceptibility to antibiotics and four phages also isolated in this work was tested. It was found that more of 90% of S. aureus isolates were not susceptible to six or more antibiotics, and 100% were resistant to penicillin, dicloxacillin, cefotaxime, ampicillin and cephalothin, and 81 and 77%, to tetracycline and cefuroxime, respectively. Fortunately, 100% of S. aureus isolates were susceptible to phages used in this work, which was detected as clear zones using specific phage. It was shown for the first time, that phages used in this study are active against pathogenic S. aureus and might be incorporated into the therapy as an important tool for the control of staphylococcal bovine mastitis, specially to antibiotic-resistant S. aureus strains isolated in farm located at the state of Guanajuato, México; and its use might be extended to other regions inside or outside the country.

Characterization and Identification of Cryptic Biopeptides in Carya illinoinensis (Wangenh K. Koch) Storage Proteins.

The objective of this research was to identify and characterize the encoded peptides present in nut storage proteins of Carya illinoinensis. It was found, through in silico prediction, proteomic analysis, and MS spectrometry, that bioactive peptides were mainly found in albumin and glutelin fractions. Glutelin was the major fraction with ~53% of the nut storage proteins containing at least 21 peptides with different putative biological activities, including antihypertensives, antioxidants, immunomodulators, protease inhibitors, and inhibitors of cell cycle progression in cancer cells. Data showed that using 50 µg/mL tryptic digests of enriched peptides obtained from nut glutelins is able to induce up to 19% of apoptosis in both HeLa and CasKi cervical cancer cells. To our knowledge, this is the first report that shows the potential value of the nut-encoded peptides to be considered as adjuvants in cancer therapies.

Cucurbitaceae Seed Protein Hydrolysates as a Potential Source of Bioactive Peptides with Functional Properties.

Seeds from Cucurbitaceae plants (squashes, pumpkins, melons, etc.) have been used both as protein-rich food ingredients and nutraceutical agents by many indigenous cultures for millennia. However, relatively little is known about the bioactive components (e.g., peptides) of the Cucurbitaceae seed proteins (CSP) and their specific effects on human health. Therefore, this paper aims to provide a comprehensive review of latest research on bioactive and functional properties of CSP isolates and hydrolysates. Enzymatic hydrolysis can introduce a series of changes to the CSP structure and improve its bioactive and functional properties, including the enhanced protein solubility over a wide range of pH values. Small-sized peptides in CSP hydrolysates seem to enhance their bioactive properties but adversely affect their functional properties. Therefore, medium degrees of hydrolysis seem to benefit the overall improvement of bioactive and functional properties of CSP hydrolysates. Among the reported bioactive properties of CSP isolates and hydrolysates, their antioxidant, antihypertensive, and antihyperglycaemic activities stand out. Therefore, they could potentially substitute synthetic antioxidants and drugs which might have adverse secondary effects on human health. CSP isolates and hydrolysates could also be implemented as functional food ingredients, thanks to their favorable amino acid composition and good emulsifying and foaming properties.

First Molecular Identification of Dirofilaria repens in a Dog Blood Sample from Guanajuato, Mexico.

Dirofilaria repens is the causative zoonotic agent of subcutaneous dirofilariosis. Although the American continent is considered currently free of D. repens infections, in the past few years there have been some reports suggesting the presence of this agent in the Americas. In Guanajuato, Mexico, there is a report of the presence of D. repens. To determine whether this parasite is really present in Guanajuato, a total of 177 EDTA blood samples from dogs were tested using the modified Knott's technique. Using this technique, we found 15 positive samples, which were then analyzed using the PCR technique, with a panfilarial set of primers and a specific primer pair for D. repens. In one sample, the expected band was present and the corresponding amplicon, sequenced, displayed a 100% identity to D. repens, confirming the presence of this exotic and zoonotic filarial species in the municipality of Silao, Guanajuato, Mexico.

Molecular detection and sensitivity to antibiotics and bacteriocins of pathogens isolated from bovine mastitis in family dairy herds of central Mexico.

Thirty-two farms (n = 535 cows) located in the state of Guanajuato, Mexico, were sampled. Pathogens from bovine subclinical mastitis (SCM) and clinical mastitis (CLM) were identified by 16S rDNA and the sensitivity to both antibiotics and bacteriocins of Bacillus thuringiensis was tested. Forty-six milk samples were selected for their positive California Mastitis Test (CMT) (≥3) and any abnormality in the udder or milk. The frequency of SCM and CLM was 39.1% and 9.3%, respectively. Averages for test day milk yield (MY), lactation number (LN), herd size (HS), and number of days in milk (DM) were 20.6 kg, 2.8 lactations, 16.7 animals, and 164.1 days, respectively. MY was dependent on dairy herd (DH), LN, HS, and DM (P < 0.01), and correlations between udder quarters from the CMT were around 0.49 (P < 0.01). Coagulase-negative staphylococci were mainly identified, as well as Staphylococcus aureus, Streptococcus uberis, Brevibacterium stationis, B. conglomeratum, and Staphylococcus agnetis. Bacterial isolates were resistant to penicillin, clindamycin, ampicillin, and cefotaxime. Bacteriocins synthesized by Bacillus thuringiensis inhibited the growth of multiantibiotic resistance bacteria such as S. agnetis, S. equorum, Streptococcus uberis, Brevibacterium stationis, and Brachybacterium conglomeratum, but they were not active against S. sciuri, a microorganism that showed an 84% resistance to antibiotics tested in this study.

Identification of differential expressed transcripts in cervical cancer of Mexican patients.

The aim of this study was to identify the gene expression profile in biopsies of patients with cervical intraepithelial neoplasia (CIN) 1, CIN 2, CIN 3, and microinvasive cancer by suppression subtractive hybridization and Southern blotting. After analyzing 1,800 cDNA clones, we found 198 upregulated genes, 166 downregulated, and no significant change of gene expression in 86 clones (p = 0.005). These results were validated by Northern blot analysis (p = 0.0001) in the identification of 28 overexpressed and 7 downregulated transcripts. We observed a set of genes related to the Notch signaling pathway that may be involved in the transformation of cervical cells and in the development to malignancy. The differentially expressed genes may provide useful information about the molecular mechanisms involved in human cervical carcinoma and as diagnostic markers.