Plasma Concentrations of Rosmarinic Acid in Patients on Antiretroviral Therapy: In Silico Exploration Based on Clinical Data.

Rosmarinic acid (RA) is a phenolic compound with antiviral properties, often encountered in dietary supplements and herbal drugs. Data on the pharmacokinetics of RA are lacking in cases of the chronic use of supplements containing this compound, and only limited data on the metabolism and distribution of RA are available. The aim of the study was to investigate the plasma levels of RA after 12 weeks of use and determine potential interactions of RA and selected antiretroviral drugs. Patients infected with human immunodeficiency virus took a supplement containing RA for 12 weeks, after which the RA concentrations in the plasma samples were analyzed. A detailed in silico analysis was conducted in order to elucidate the potential interactions between RA and the drugs efavirenz, darunavir and raltegravir. It was found that RA can be detected in patients' plasma samples, mainly in the form of sulphoglucuronide. The potential interactions are suggested on the level of liver metabolizing enzymes and efflux P-glycoprotein, with RA competing with antiretroviral drugs as a substrate in metabolism and distribution systems. The present study suggests that the simultaneous use of RA and antiretroviral therapy (containing efavirenz, darunavir or raltegravir) may affect the plasma levels of RA after prolonged supplementation.

Assessment of antibiotic use patterns in bovine mastitis treatment in the dairy sector in Serbia.

The dairy sector is essential for the world's food systems, playing a vital role in agricultural production. Yet, mastitis, an inflammation of the mammary gland, affects a considerable portion of the dairy cow population annually, leading to reduced productivity and sustainability. Insight into therapeutic protocols is essential for a better understanding of the situation on farms with the aim of developing new or harmonizing existing protocols in the mastitis treatment. This study conducted on dairy farm in Serbia aimed to explore the most commonly used antibiotics during bovine mastitis treatment and their association with daily milk yield. Data from 100 lactating cows, including therapeutic protocols, lactation numbers, mastitis episodes, milk yield, and antibiotic usage, were obtained from a computerized database and statistically analysed. The results demonstrated a high prevalence of narrow-spectrum antibiotics usage, with the majority of cows treated with combination therapy. The route of antibiotic application did not significantly impact daily milk yield, while treatment duration had no discernible effect. Nonetheless, specific antibiotics were associated with milk yield variations, suggesting the need for careful antibiotic selection and management in mastitis therapy. These findings emphasize the importance of prudent antibiotic use to safeguard animal health and long-term milk production sustainability.

Comparative Health Risk Assessment of Heated Tobacco Products versus Conventional Cigarettes.

BACKGROUND: Smoking is the largest preventable cause of death in the world with around eight million estimated premature deaths per year. In response to the harmful effects of conventional tobacco products the tobacco industry has launched a new type of products called Heated tobacco products (HTP) and e-cigarettes, which are considered safer for human health than conventional cigarettes. MATERIALS AND METHODS: The research was conducted by searching the scientific literature using platforms "Google scholar," "PubMed" and "Science Direct." To compare the non-carcinogenic and carcinogenic risk of HTP products and conventional cigarettes, margin of exposure and lifetime excess cancer risk were calculated using data obtained from a scientific literature search.HTP products have shown a reduced risk to human health compared to the conventional cigarettes, although they still contain compounds that can be dangerous to human health. There is not enough data obtained from independent studies that could safely indicate that these reduced amounts of toxic chemical entities in the composition of HTP do not induce any harmful effect. CONCLUSION: Further research is needed to determine the harmful effects of HTP aerosol, as well as to tighten the legislation that would limit the production, import and distribution of these products worldwide until their safety for human health is confirmed with a sufficient number of transparent and representative results obtained in independent scientific studies.

Biological Activity of Genus Hypericum Sect. Hypericum Species-H. tetrapterum, H. maculatum subsp. immaculatum, H. triquetrifolium.

St. John's wort (Hypericum perforatum, Hypericaceae) has long been used in traditional medicine as a potent remedy, while many other species of this genus have not been thoroughly investigated. The study aimed to detect the biological activity, including antioxidant, antihyperglycemic, anticholinergic, antimicrobial and monoaminoxidase inhibitory potential, of water-alcoholic extracts of three species autochthonous for Serbia and Greece from plant genus Hypericum (section Hypericum-H. tetrapterum, H. maculatum ssp. immaculatum and H. triquetrifolium), followed by phytochemical profiling. The highest amount of phenolics was recorded in H. maculatum subsp. immaculatum extract, while the highest abundance of flavonoids was characteristic of H. tetrapterum extract. Hypericin and hyperforin, quercetin, and its flavonoid, rutin, were present in all of the evaluated species. The evaluated species were good scavengers of DPPH, OH and NO radicals, as well as potent reducers of ferric ions in FRAP assay. Furthermore, the evaluated species were shown as potent inhibitors of monoaminoxidase A and α-glucosidase and modest inhibitors of acetylcholinesterase, monoaminoxidase B and α-amylase. No anti-Candida activity was recorded, but the extracts were effective against MRSA Staphylococcus aureus and Enterococcus sp., as well as against Proteus mirabilis. The obtained results strongly highlight the need for further in vivo studies in order to better define the potential of the medicinal application of the studied species.

Phytochemistry and Antihyperglycemic Potential of Cistus salviifolius L., Cistaceae.

Cistus salviifolius has been previously reported as a traditional remedy for hyperglycemia. However, the plant has been scarcely investigated from scientific point of view. Thus, the aim was to examine the chemical composition and to evaluate its antioxidant and antihyperglycemic potential in vitro. Aqueous and ethanolic extracts were evaluated for total phenolic, tannin, and flavonoid content using spectrophotometric methods. Detailed chemical characterization was performed by high-performance liquid chromatography (HPLC-DAD). The volatile organic compounds (VOCs) profile was assessed by gas chromatography technique. The potential in diabetes treatment was evaluated through tests of free radicals neutralization, inhibition of lipid peroxidation process, and test of ferric ion reduction; activity in tests of inhibition of α-amylase, α-glucosidase and dipeptidyl peptidase-4 was also evaluated. High content of phenolics (majority being tannins) was detected; detailed HPLC analysis revealed high content of gallic acid, followed by rutin, chlorogenic and caffeic acids. The VOCs analysis determined sesquiterpene hydrocarbons and oxygenated sesquiterpenes as the main groups of compounds. The assays classified extracts as potent neutralizers of 2,2-diphenyl-1-picrylhydrazil and nitroso radicals formation and potent inhibitors of α-amylase and α-glucosidase. In conclusion, Cistus salviifolius represents a rich source of phenolics and essential oil with sesquiterpenes. The established results suggested its promising antioxidant and antihyperglycemic activities.

The Evaluation of Healing Properties of Galium verum-Based Oral Gel in Aphthous Stomatitis in Rats.

Although oral ulcers represent one of the most frequent oral mucosal diseases, the available treatment is not sufficient to provide complete ulcer recovery without side-effects. Therefore, the aim of our study was to prepare a mucoadhesive oral gel based on Galium verum ethanol extract (GVL gel) and reveal its healing effects in the model of aphthous stomatitis in rats. Rats with oral ulcers were divided into the following groups: control (untreated), gel base (ulcer was treated with the gel base, three times per day for 10 days), and GVL gel group (the ulcer was treated with GVL gel in the same way as the gel base). Animals from each group were sacrificed on days 0, 3, 6, and 10 for collecting blood and ulcer tissue samples. Healing properties of oral gel were determined by clinical evaluation, as well as biochemical and histopathological examinations. Our findings suggest a significant decrease in the ulcer size in GVL gel group, with healing effects achieved through the alleviation of oxidative stress, reduction in COX-2 immunopositivity, and increase in collagen content in buccal tissue. Significant ulcer repairing potential of GVL gel highlights this oral mucoadhesive gel as a promising tool for prevention and treatment of RAS.

Rosmarinic Acid-Human Pharmacokinetics and Health Benefits.

Rosmarinic acid is a phenolic compound commonly found in the Lamiaceae (Labiateae) plant species. It is considered responsible for a wide spectrum of biological and pharmacological activities of plants containing this compound. The aim of the current review is to present the fate of rosmarinic acid inside the human body, explained through pharmacokinetic steps and to briefly present the health benefits of RA. Pharmacokinetics was at first studied in animal models, but several studies were conducted in humans as well. This compound can be applied topically, pulmonary, intranasally, and via intravenous infusion. However, peroral application is the main route of entry into the human body. Presumably, it is mainly metabolized by the gut microflora, providing simple, more easily absorbed phenolic units. Inside the body, the rosmarinic acid molecule undergoes structural changes, as well as conjugation reactions. Renal excretion represents the main path of elimination. Previously conducted studies reported no serious adverse effects of herbal remedies containing RA, as well as their positive effects on human health. In addition to in vitro studies, clinical investigations suggested its benefits in dermatological, allergic, and osteoarthritic disorders, as well as for improving cognitive performance and in metabolic syndrome treatment. Future studies should investigate the kinetics during long-term application in patients who would have potential benefits from RA usage. Pharmaceutical formulations designed to prevent the fast metabolism of RA and allow its penetration into other compartments of the human body are also interesting topics for future research.

Comparison of mechanical, chemical and physical human models of in vivo skin damage: Randomized controlled trial.

INTRODUCTION: Human in vivo models of skin damage were often used in research of cutaneous disorders. The most commonly used models were tape-stripping as mechanical, sodium lauryl sulphate-induced irritation as chemical and ultraviolet radiation as physical damage model. In regard to differences between models, they were expected to have different responses to damage and recovery, with unique skin parameters' changes over time. OBJECTIVE: The aim was to compare skin parameters in three different skin damage models on the same anatomical location, with and without topical treatment. METHODS: Four test sites on each forearm were randomly assigned to three skin damage models with the fourth sites on each forearm chosen as a control, undamaged site. Skin parameters were assessed using non-invasive methods. RESULTS: Sodium lauryl sulphate irritation caused the strongest damage with delayed reaction to the irritant. Tape stripping leads to highest initial skin barrier disruption but afterwards it showed the fastest skin recovery. Ultraviolet radiation did not affect skin barrier function, but it elevated skin erythema and melanin level. Tested preparation did not lead to changes in measured parameters. CONCLUSION: The skin of the participants had different response to three skin damage models with distinct changes of skin parameters and recovery. The trial was registered at ClinicalTrials.gov under the identifier NCT03783819.

Garlic burn injuries- a systematic review of reported cases.

Medicinal plants have many beneficial effects on human health. Garlic (Allium sativum, Alliaceae) is one of the most famous herbal species, used for various diseases and conditions. Unfortunately, garlic is also associated with adverse effects, including cutaneous manifestations. In this review, burn injuries caused by application of raw garlic are reported. Searching through PubMed, Google Scholar and ResearchGate, a total of 32 articles with 39 patients were found. Demographics of patients, reasons for garlic use, details on garlic application, as well as description of burns and its treatment are thoroughly described and discussed. In most of the cases, garlic caused second-degree burns, although some circumstances can cause formation of necrotic tissue. Various body parts were affected, legs being most common. The chemistry of garlic is also presented, with focus on volatile organic sulfur compounds, which also seem to be responsible for burns formation. Treatment of garlic burns was mainly symptomatic, and various types of drugs were used. Although not commonly expected, garlic should be taken into consideration as causative agents of burns by treating doctors, and patients should be advised against application of fresh garlic onto skin and mucosa.

Chemical Diversity of Volatile Compounds of Mints from Southern Part of Pannonian Plain and Balkan Peninsula - New Data.

Mints are the most popular economic and traditional herbs. The aim of this article was chemical characterization of volatile compounds from wild populations of Mentha aquatica, M. arvensis, M. longifolia, M. microphylla, M. pulegium, M. spicata, M.×dumetorum, M.×gentillis and M.×verticillata, as well as cultivated samples of M. spicata, M.×piperita 'Alba' and M.×piperita 'Crispa'. Analyses were performed directly from dried aerial parts (herb) of collected samples by headspace gas chromatography-mass spectrometry. In total 54 compounds were detected, representing from 89.99 % to 99.66 % of volatile fractions of all investigated samples. The recorded volatiles were primarily monoterpene hydrocarbons and oxygenated monoterpenes, while oxygenated aromatic monoterpenes, sesquiterpene hydrocarbons and aliphatic compounds were present in lower concentrations in analyzed samples. The major components were linalool, limonene, 1,8-cineol, α-terpinyl acetate, pulegone, ß-pinene and menthol. The cluster analysis revealed five main groups or chemotypes according to qualitative and quantitative content of volatiles, as well as similarities among samples. These results contribute to the knowledge on the mints chemistry in Pannonian Plain and Balkan Peninsula.

St. John's Wort Herbal Teas - Biological Potential and Chemometric Approach to Quality Control.

St. John's wort has been used for centuries in traditional medicine of many cultures, and nowadays it is well-known as a clinically important antidepressant drug. Considering the rising market demand for Hyperici herba, quality control of crude drug is of paramount importance. In this paper we performed HPLC-DAD chemical profiling of St. John's wort tea samples obtained at local markets, pharmacies and health food stores in the Balkan Peninsula countries, Austria and Turkey. Furthermore, water alcoholic extracts of the collected samples were evaluated in terms of their antioxidant potential, as well as the ability to inhibit biologically important enzymes such as acetylcholinesterase, monoamine oxidases A and B (MAO-A and MAO-B), α-amylase and α-glucosidase. Significant variability within the samples in the quantities of hypericin, hyperforin, rutin, quercetin, gallic, chlorogenic, caffeic and p-hydroxybenzoic acid was noticed. Chemotaxonomic modelling enabled the identification of three clusters of samples based on the quantities of rutin, hypericin and hyperforin. Generally, the extracts exhibited a significant potential to inhibit MAO-A (median IC50 = 10.01 µg/mL) and α-glucosidase (median IC50 = 12.40 µg/mL). The results of antioxidant potential evaluation suggest strong neutralization of hydroxyl and nitroso radicals, but moderate inhibition of lipid peroxidation process. Overall, the conducted study emphasizes the possibility of St. John's wort teas quality control, taking into account unavoidable variability of chemical profile as well. The stated is of great importance when grinding degree of herbal material excludes taxonomic identification of biological origin as an option within quality control.

Health risk assessment for pediatric population associated with ethanol and selected residual solvents in herbal based products.

In this study, 48 herbal based products (41 for the pediatric population) were analyzed for the presence of ethanol and residual solvents. Ethanol was not detected in only 12% of the products designed for infants or toddlers aged under 2, and not quantified in only 5 of 14 'alcohol free' products. Actual content was higher than labeled in six out of 11 samples with specified ethanol quantity. WHO proposed requirement for ethanol content in products intended for use in children under the age of 6 (<0.5%) was not met by as many as 26 samples. Furthermore, calculated blood alcohol levels in children exceeded the relevant toxicological levels for nine samples following a single dose, and for one sample in case of accidental poisoning with the entire package. Regarding the residual solvents, acetone, 1-propanol and 1-butanol were not quantified, 2-propanol was found in two samples in low concentrations, whereas methanol intake via one of the samples exceeded the permitted level for children. The obtained results revealed a significant health concern for the pediatric population due to ethanol intake via herbal based products, calling for the establishment of strict guidelines for ethanol content and labeling.

Potential of selected Lamiaceae plants in anti(retro)viral therapy.

Constant search for new drugs with antiviral properties often extends to products of natural origin. Lamiaceae is one of the most important herbal families, well known for various biological and medicinal effects of a variety of aromatic spices, including thyme, mint, oregano, basil, sage, savory, rosemary, self-heal, hyssop, lemon balm and many others. The paper provides a review of antiviral potential of previously mentioned plants which has been demonstrated so far, with special emphasis on anti-HIV properties. Relevant articles were compiled by searching plant names combined with keywords describing antiviral activity. The antiviral effect is direct, with prominent activity against enveloped viral species. Initial stages of the viral life cycle are the most affected, as these plants appear to be targeting mainly viral structures responsible for attachment to target cells. In case of HIV, there is some activity against key enzymes in the viral life cycle. Even in the case of drug resistance, there is an equal susceptibility to applied herbal preparations. Some in vivo experiments suggest that use of Lamiaceae representatives could help in prevention and treatment of some viral diseases. A possible reduction of side effects of diseases and conventional drug therapy are also some aspects worth further investigations.

The Phenolics of the Ornithogalum umbellatum L. (Hyacinthaceae): Phytochemical and Ecological Characterization.

Ornithogalum umbellatum L. is a widely distributed species in Europe, that exhibits considerable variability at ecological, morphological, anatomical, and karyological level. Previous reports of the chemical investigations among Ornithogalum species indicate significant diversity of the secondary metabolites, as well. Knowing that environment affects the phenolic composition in plants to a large extent, the main objective of the research was to define relationship between phytochemical and ecological characters. To estimate an environmental influence on these results, plant material was collected at four habitats that differ in ecological factors and belong to two biogeographical regions: the Balkan Peninsula and the Pannonian Plane. Measured phytochemical characters are yield of dry extract, total phenolic and flavonoid contents, the presence of selected phenolic compounds as well as free-radical scavenging activity (neutralization of DPPH and OH radicals). Results revealed that all analyzed phytochemical parameters differ between investigated O. umbellatum samples. The moisture level of habitat has the highest correlation, either positive or negative, with most of phytochemical characters, and is followed by temperature and soil reaction. Light intensity and nitrogen level have mostly moderate correlation coefficient with phytochemical characters. More complex correlation is revealed between ecological factors and nine phenolic compounds, with three observed patterns of relationship.

Biochemical characterization of Helichrysum italicum (Roth) G.Don subsp. italicum (Asteraceae) from Montenegro: phytochemical screening, chemotaxonomy, and antioxidant properties.

The chemical composition and antioxidant properties of the essential oil and EtOH extract of immortelle (Helichrysum italicum (Roth) G.Don subsp. italicum, Asteraceae) collected in Montenegro were evaluated. The essential oil was characterized by GC/MS analysis, and the content of total phenolics and flavonoids in the EtOH extract was determined using the FolinCiocalteu reagent. The free-radical-scavenging capacity (RSC) of both the essential oil and the EtOH extract was assessed with the 2,2-diphenyl-1-pycrylhydrazyl (DPPH) method. Moreover, the inhibition of hydroxyl radical ((.) OH) generation by the EtOH extract of immortelle was evaluated for the first time here. Neryl acetate (28.2%) and γ-curcumene (18.8%) were the main compounds in the essential oil, followed by neryl propionate (9.1%) and ar-curcumene (8.3%). The chemical composition of the oils of the examined and additional 16 selected Helichrysum italicum taxa described in literature were compared using principal component (PCA) and cluster (CA) analyses. The results of the statistical analyses implied the occurrence of at least four different main and three subchemotypes of essential oils. Considering the antioxidant properties, the EtOH extract of immortelle exhibited similar potential as propyl gallate and quercetin, while the essential oil exhibited relatively weak DPPH(.) -scavenging capacity.

The Potential of Hemp Extracts to Modify the Course of Oxidative-Stress Related Conditions.

The leaves of industrial hemp, which have long been considered as a waste product, have been proven to contain numerous compounds that possess potential biological activity. One of the most interesting groups of compounds present are polyphenolic compounds, which, due to their specific structure, have a pronounced antioxidant and antihyperglycemic potential. This study aimed to detect biological activity, including antioxidant and antihyperglycemic potential, of water and water-alcoholic extracts of five commercially available hemp teas, followed by phytochemical profiling. Hemp aqueous and ethanolic extracts demonstrate potent antioxidant properties. Ethanol extracts are better scavengers of DPPH• and OH•, while aqueous extracts neutralize NO• better. Both types of extracts exhibit antioxidant potential in the catalase test and moderate XOD inhibition. Furthermore, aqueous extracts are potent α-amylase inhibitors, while ethanolic extracts demonstrate stronger anti-α-glucosidase activity, suggesting therapeutic potential for chronic diseases like insulin resistance or diabetes. Further detailed chemical characterization and in vivo studies are needed to validate these findings.

Impact of origin and biological source on chemical composition, anticholinesterase and antioxidant properties of some St. John's wort species (Hypericum spp., Hypericaceae) from the Central Balkans.

The study shows the influence of the origin of plant material and biological source on the in vitro antioxidant (neutralization of DPPH and OH radical, nitric oxide, and inhibition of lipid peroxidation) and anticholinesterase activity of chemically characterized and quantified ethanol extracts of ten St. John's wort samples. The investigated samples were: five Hypericum perforatum species representatives collected at different localities, one commercial sample of Hyperici herba purchased at a local market and four Hypericum species autochtonous to the Balkan Peninsula (H. maculatum subsp. immaculatum, H. olympicum, H. richeri subsp. grisebachii and H. barbatum). All the examined extracts exhibited notable antioxidant potential, but in most of the cases indigenous Hypericum species expressed stronger effects compared to the original source of the drug, H. perforatum. The changes in the content of phenolic compounds, especially flavonoids, hyperforin and hypericin, related to the source of the drug affected the investigated activities. Since all of the investigated species have shown prominent inhibition of acetylcholinesterase in vitro activity, they could be further investigated as potential substances in preventing of Alzheimer's disease.

Health risk assessment of ethyl carbamate in fruit spirits.

Alcoholic beverages are multicomponent mixtures containing several carcinogenic compounds, including ethyl carbamate, which was evaluated in fruit spirits against the limit proposed by the International Alliance for Responsible Drinking (IARD) and then in terms of associated health. It was above the limit in 40% of the samples and statistically higher in stone fruit spirits. The MOE values below the limit were obtained for 49% and 98% of the samples at average and chronic heavy consumption by men, while for women the limit was reached by 19% and 96% of the samples, respectively. The lifetime cancer risk (LCR) approach based on the oral slope factor indicated risk concern for 96% of the samples at average consumption, regardless of gender, while LCR based on virtually safe dose indicated 25% and 69% of the samples posing a risk to women's and men's health, respectively. Preventive measures during spirit production and legal regulation of ethyl carbamate should be considered.

PREDICTIVE POWER OF OXIDATIVE STRESS BIOMARKERS IN RECURRENCE AND SURVIVAL IN ADVANCED CERVICAL CANCER.

The aim of our study was to measure the levels of 8-hydroxy-2-deoxyguanosine, malondialdehyde, and antioxidant enzymes in patients with locally advanced cervical cancer prior to treatment to determine how these evaluated biomarkers are associated with cervical cancer recurrence and to estimate their potential in further research and clinical use. MATERIALS AND METHODS: The study included 45 female patients with newly diagnosed advanced cervical cancer who underwent concomitant chemoradiotherapy. The blood and urine samples were collected prior to treatment, between December 2013 and April 2016, and subsequent laboratory analysis was performed. After the medium follow-up of 29 months, the patients were divided into 3 groups according to the time of disease recurrence. A statistical analysis was performed in order to evaluate the relationship between the previously measured biomarkers and recurrence. RESULTS: Taken individually, the parameters of oxidative stress did not reveal significant differences between the three groups in our study. Nevertheless, the catalase and glutathione S-transferase activities were the best predictors of the recurrence. Based on the activities of these two oxidative enzymes, it was possible to separate the group of patients without recurrence after follow-up from the other two groups of patients with recurrent disease. CONCLUSIONS: The parameters of oxidative stress have a certain predictive value on the outcome of patients with advanced cervical cancer after concomitant chemo-radiotherapy.

Assessment of Mastitis Patterns in Serbian Dairy Cows: Blood Serum Metabolic Profile and Milk Composition Parameters.

Mastitis is one of the most important diseases in dairy cows, leading to substantial economic losses associated with decreased milk production and quality. Early detection of changes in metabolic and milk parameters is crucial for maintaining animal welfare and milk quality. This study aimed to detect patterns in metabolic and milk composition parameters in Serbian dairy cows affected by mastitis. It also examined the relationship between these factors in cows with clinical and subclinical mastitis, as well as in healthy cows. This study included 60 Holstein-Friesian cows with the same body score condition that were in the same lactation phase. They were divided into three groups of 20: clinical and subclinical mastitis and a control group of healthy cows. The categorization was based on clinical udder health and the California mastitis test. Blood serum metabolic profiles were measured using a Rayto spectrophotometer (Shenzhen, China), and milk composition was determined using MilcoScanTM (Foss, Hilleroed, Denmark) and FossomaticTM (Foss, Hilleroed, Denmark) instruments. Significant increases in non-esterified fatty acids (NEFAs), beta-hydroxybutyrate (BHB), total protein, globulin, urea, total bilirubin, magnesium, and enzyme activity were noted in mastitis-affected cows compared to healthy ones. Additionally, mastitis-affected cows had higher total protein and globulin levels and increased somatic cell counts (SCCs), while albumin concentrations were decreased. Furthermore, a negative correlation between total protein and lactose suggested inflammation leading to reduced lactose levels due to cell damage, infection, and lactose use by mastitis pathogens. Hence, indicators of the energy and protein status of the metabolic profile, together with the chemical composition of milk, may be significant diagnostic tools for detecting, monitoring, and predicting the outcome of mastitis in cows.