Caffeine Concentrations Research Articles

Caffeine Supplementation in Semen Extender Protects Buffalo Spermatozoa from Cryodamage

Caffeine is widely known for its phosphodiesterase enzymes inhibiting, and its powerful reactive oxygen species scavenging property. This study aimed to evaluate the effect of caffeine supplementation @ 0 mM, 1 mM, 3 mM and 5 mM in AndroMed® extender on cryopreservation of Jaffarabadi buffalo semen. Twenty-four semen ejaculates (6/bull) with >70% initial sperm motility were obtained from four mature Jaffarabadi bulls using AV. The ejaculates were split-diluted in an egg-yolk-free AndroMed® extender supplemented with different concentrations of caffeine and were cryopreserved using a standard protocol. The semen samples were evaluated for sperm quality parameters as well as oxidative stress parameters, viz., lipid peroxidation, and Glutathione-S-transferase (GST) enzyme activity in seminal plasma at pre-freeze (after equilibration) and post-thaw stage. The levels of caffeine had a significant effect on all these parameters, except sperm abnormalities, at both pre-freeze and post-thaw stages. Supplementation of caffeine in semen extender at 1 mM and 3 mM concentration showed a significant (p<0.05) increase in post-thawed sperm motility (62.04 ± 0.84, 61.25 ± 1.01%), viability (64.21 ± 0.88, 64.25 ± 0.84%), acrosome integrity (58.08 ± 1.08, 57.30 ± 0.93%) and plasma membrane integrity (52.75 ± 0.89, 52.71 ± 0.74%) and a significant (p<0.05) decrease in oxidative damage as evident by lower lipid peroxidation (MDA 7.62 ± 0.41, 7.80 ± 0.70 μM) and GST enzyme activity (31.15 ± 1.36, 29.54 ± 0.54 nmol CDNB/mL/min) as compared to control and 5 mM caffeine. The study concludes that the post-thaw quality of frozen semen of Jaffarabadi buffalo bulls improves significantly with decreased oxidative stress if the AndroMed® extender is supplemented with 1 and 3 mM concentration of caffeine over control.

Long-Term Traditional Fertilization Alters Tea Garden Soil Properties and Tea Leaf Quality in Bangladesh

Soil acidity is one of the major soil-degradation events throughout the world, and the long-term application of nitrogenous fertilizers is thought to be a main cause of soil acidity. In the present experiment, we collected soil and tea (Camellia sinensis L.) leaf samples from five representative tea gardens in Bangladesh and evaluated soil nutrient pools and biochemical properties of tea leaves. The results showed that there was a negative relationship between soil pH and the amount of applied nitrogenous fertilizers. Moreover, continuous application of traditional fertilizers over twenty-five years promoted not only the deficiency of phosphorus (P) and mineral-based cations, such as potassium (K+), calcium (Ca2+), and magnesium (Mg2+), but also increased manganese (Mn2+) and aluminum (Al3+) toxicity in soils, which suppressed the yield and quality of tea. Crucially, tea leaf production remained almost similar (average 1079.77 kg ha−1) from 1995 to 2015, while the application doses of urea, TSP, and MoP increased by 24.69%, 18.92%, and 16.67%, respectively, in garden soils. However, the pH value of soil declined up to 24% from 1992 to 2020 in the tested gardens. Consequently, the availability of K+, P, Ca2+, and Mg2+ decreased by 56%, 25%, 55%, and 49%, respectively, in those tea garden soils. In addition, the quality of tea leaves was severely affected, as evident by the reduced levels of total flavonoids, polyphenols, soluble solids, vitamin C, vitamin B1, and vitamin B2. Moreover, free-radical scavenging activity (DPPH), caffeine, and tannin concentration were increased in tea leaves, which indicated that tea plants were potentially being stressed. Therefore, we study concluded that long-term application of traditional nitrogenous fertilizers can be an important regulator of lowering garden soil pH, which reduces native soil nutrient pools and thereby the yield and quality of tea leaves.

Population pharmacokinetic modeling of caffeine in preterm infants with apnea of prematurity: New findings from concomitant erythromycin and AHR genetic polymorphisms

Current standard-dose caffeine therapy results in significant intersubject variability. The aims of this study were to develop and evaluate population pharmacokinetic (PPK) models of caffeine in preterm infants through comprehensive screening of covariates and then to propose model-informed precision dosing of caffeine for this population. A total of 129 caffeine concentrations from 96 premature neonates were incorporated into this study. Comprehensive medical record and genotype data of these neonates were collected for analysis. PPK modeling was performed by a nonlinear mixed effects modeling program (NONMEM). Final models based on the current weight (CW) or body surface area (BSA) were evaluated via multiple graphic and statistical methods. The model-informed dosing regimen was performed through Monte Carlo simulations. In addition to CW or BSA, postnatal age, coadministration with erythromycin (ERY), and aryl hydrocarbon receptor coding gene (AHR) variant (rs2158041) were incorporated into the final PPK models. Multiple evaluation results showed satisfactory prediction performance and stability of the CW- and BSA-based models. Monte Carlo simulations demonstrated that trough concentrations of caffeine in preterm infants would be affected by concomitant ERY therapy and rs2158041 under varying dose regimens. For the first time, ERY and rs2158041 were found to be associated with the clearance of caffeine in premature infants. Similar predictive performance and stability were obtained for both CW- and BSA-based PPK models. These findings provide novel insights into caffeine precision therapy for preterm infants.

Pharmaceutically active compounds in biotic and abiotic media of rivers receiving urban sewage: Concentrations, bioaccumulation and ecological risk

Pharmaceutically active compounds (PhACs) are recognized as posing health risks to aquatic ecosystems. The occurrence characteristics and ecological effects of 15 PhACs were investigated in the multiphase media of Yangtze River (Nanjing area, China) during wet and dry seasons. All 15 PhACs were detected in the traditionally dissolved phase, suspended particulate matter, sediment and fish, with total concentrations of PhACs (ΣPhACs) of 64.75–366.20 ng/L, 18.50–69.40 ng/g, 3.29–23.30 ng/g and 0.00–176.44 ng/g, respectively. Ibuprofen (IPF), roxithromycin (ROX) and caffeine (CFI) were the dominant PhACs in all of the environmental media. As the main carrier for PhACs, the colloids contributed 8.60%− 77.04% of PhACs in the traditionally dissolved phase, and distribution coefficients (log Kcol=6.38–7.95) of PhACs in colloidal phase were significantly higher than those in SPM and sediment. The bioaccumulation potential of ΣPhACs in the wild fish tissues was generally in order as follows: brain > liver > kidney > gill > blood > intestines > muscle > bile, which was positively correlated with their log Kow. The concentrations of CFI, ROX and IPF in the muscle, gill and liver can be predicted by their log Kow and pollution levels in the blood, and the concentrations of PhACs in the muscle of benthic fish were much higher than those in pelagic fish. The bioaccumulation factors of ROX, fluoxetine and bezafibrate in intestines, brain and blood showed strong enrichment capacity (>5000). In the risk assessment, the acute risk level of PhACs to aquatic organisms decreases gradually with the trophic level, while the trend of chronic risk was the opposite. The chronic mixture risk of PhACs to fish was > 1 at all sampling sites, 59.79% of mixed risk quotient were provided by clofibric acid, diclofenac, erythromycin and ketoconazole. These results indicated that PhAC can cause harmful health effects to sensitive organisms both individually and mixedly.

Characterization of Extracts of Coffee Leaves (Coffea arabica L.) by Spectroscopic and Chromatographic/Spectrometric Techniques.

Coffea arabica L. leaves represent a viable alternative to the canonical matrices used for preparation of beverages, such as tea leaves and grounded coffee beans. Coffee leaves infusions are rich in antioxidant phenolic compounds and have a lower concentration of caffeine. Due to increasing interest in this field, a complete study of the bioactive compounds as chlorogenic acids, xanthones and alkaloids is noteworthy. C. arabica leaves were subjected to ultrasound-assisted extraction, and the extracts were studied via nuclear magnetic resonance spectroscopy (NMR) and chromatographic techniques coupled with mass spectrometry (HPLC-MSn) to identify and quantify the secondary metabolites profile through an untargeted data dependent approach. A quantitative analysis was performed for the major components—chlorogenic acids, mangiferin, caffeine and trigonelline—via HPLC-MS in Single Ion Monitoring (SIM) mode. In total, 39 compounds were identified. The presence of these bioactive compounds proved the strong potential of C. arabica leaves as functional food and as an alternative to classic infused beverages.

Determination of caffeine and paraxanthine concentrations in urine samples of male clients in three Syrian gymnasiums

The aim of this research is to measure caffeine levels in urine samples of random Syrian gymgoers, and to determine the paraxanthine/caffeine ratio as a mean to access CYP1A2 activity. Urine samples were collected from 60 male clients in three different gymnasiums. All participants completed a detailed questionnaire about their lifestyle habits and medical history. Caffeine and paraxanthine were extracted from urine using solid-phase extraction (Strata C18), then analyzed by HPLC-UV (Knauer-Germany). Antipyrine was used as internal standard. Most clients stated they consumed multiple cups of at least two different caffeinated drinks directly before going to the gym (coffee, tea, cola, energy drink). Caffeine concentrations in urine ranged between 2.5 and 34.23 mcg/mL. More than 60% of samples were above the 12 mcg/mL acceptable limit by the WADA/IOC. Paraxanthine concentrations in urine ranged between 1.124 and 29.141 mcg/mL. The paraxanthine to caffeine ratio was between 0.0673 and 1.869. The higher ratios were associated with taking three medications that affect CYP1A2 activity, such as ciprofloxcacin, alluporinol, rifampcin, mexitillin, or acyclovir. No correlation was found between paraxanthine/caffeine ratio, and smoking. caffeine consumption is high among Syrian gymgoers, and its level in urine exceeded the recommended threshold of 12 mcg/mL. Although it is permitted by the WADA. The paraxanthine/caffeine ratio was in accordance with other published studies. CYP1A2 activity in participants may be affected by age, medications, or the presence of a liver or kidney disease. In conclusion, caffeine intake should be monitored in gym clients, not only for doping issues, but also for health and safety concerns.

Removal of Emergent Pollutants by a Vertical Flow Constructed Wetland with Vetiveria Zizanioides: A Case Study for Caffeine

This work evaluated caffeine removal in a vertical flow constructed wetland (VFCW), planted with Vetiveria zizanioides. The feeding was continuous (synthetic influent: mineral medium and caffeine) to reduce the concentration variations in the bed. Two influent concentrations (0.75 ± 1.0 mg·L−1 and 1.5 ± 1.0 mg·L−1) were used with a constant hydraulic load (100 ± 10 L·m−2·d−1). Plant growth was monitored weekly, and characterization was carried out to determine the levels of chlorophyll a and b, pigments and carotenoids, and nutrients (nitrogen, phosphorus, magnesium, calcium, sodium and potassium). HPLC-MS was used to determine the caffeine concentration. The caffeine removal efficiency reached averages of 93 ± 1% and 87 ± 1% (retention time: 6.23 ± 0.23 hours). The caffeine contents in the influent did not affect chlorophyll a, total chlorophyll or carotenoids, and an increasing trend throughout the tests was observed. Sodium and potassium contents also showed an increase with higher caffeine concentration. This preliminary study showed that removing caffeine from wastewater using a VFCW is promising.
 Keywords: wastewater, emergent pollutants, caffeine, vertical flow constructed wetlands, Vetiveria zizanioides

Exploring the Quality and Application Potential of the Remaining Tea Stems after the Postharvest Tea Leaves: The Example of Lu'an Guapian Tea (Camellia sinensis L.).

Lu’an Guapian tea is produced through the processing of only leaves, with the stems and buds discarded, but stems constitute a large proportion of the tea harvest. To test the usability of tea stems, we compared the physicochemical properties of tea leaves and stems from the same growth period as well as the taste of their infusions. The leaves contained higher concentrations of polyphenols and caffeine and had a stronger taste. The tea stems contained higher concentrations of free amino acids and soluble sugars and were richer in umami and sweet flavors. In addition, more tender tea stems had higher concentrations of polyphenols, caffeine, and free amino acids, and their infusions had more refreshing and sweeter tastes. Furthermore, crude fiber content increased as stem tenderness decreased. In summary, tea stems are rich in phytochemical components and flavor, and these properties increased with tenderness. This provides a theoretical basis for the high-value utilization of tea stems.

Assessment of caffeine neurotoxicity using novel biomarkers of neural function in SH-SY5Y cells – Is there a need for environmental concern?

Worldwide usage of caffeine results in its constant release into the aquatic environment and growing concerns related to associated risks. We assessed (neuro)toxicity of environmentally relevant concentrations of caffeine, using novel biomarkers of neural function in SH-SY5Y cells and markers of general toxicity also in HepG2 cells. The RQ-PCR analyses showed that caffeine disturbs the expression of genes encoding several key elements of neurotransmitter pathways, with the most prominent responses observed for serotonin receptor 3A, dopamine receptor D2, monoamine oxidase B and GABA-transaminase. Expression of genes encoding synaptotagmin 10 involved in exocytosis of neurotransmitters and ATPase Na+/K+ transporting subunit alpha 3 was also disturbed. Caffeine stimulated the activity of monoamine oxidase, while cytotoxicity and effects on mitochondrial membrane potential were not observed. Our study points out the new possible molecular targets of caffeine and suggests that the raising concerns related to its growing environmental presence are justified.

Impact of agricultural runoff and domestic sewage discharge on the spatial-temporal occurrence of emerging contaminants in an urban stream in São Paulo, Brazil.

Ribeirão das Pedras, a 10-km-long stream from the source to mouth, is part of a predominantly urban catchment located in Campinas metropolitan area in the state of São Paulo, Brazil, and it is also surrounded by sugarcane farms. Monthly sampling of 31 selected emerging contaminants (ECs) was conducted for 1year (October 2018 to October 2019) in five points, including the spring, agricultural, and urban areas, to assess the dynamics and impact of ECs on the stream. The ECs were quantified using LC-MS/MS analysis. Out of the 31 ECs monitored in this study, 13 were detected in the Ribeirão das Pedras catchment, which were mainly pesticides and caffeine. Eight ECs (hexazinone, malathion, desethylatrazine (DEA), desisopropylatrazine (DIA), fipronil, ametryn, 2-hidroxyatrazine, and diuron) were detected with risk quotients higher than 1, indicating some level of environmental concern. Statistical analyses showed that caffeine, hexazinone, atrazine, DEA, and DIA were the most statistically important contaminants in temporal analysis, with caffeine concentrations varying randomly. Hexazinone, atrazine, DIA, and DEA concentrations increased from November 2018 to January 2019, and atrazine, hexazinone, and DEA concentrations increased from June 2019 to September 2019. Spatial analysis indicates that the spring of Ribeirão das Pedras is the only statistically different sampling point, with lower concentrations of EC. Points 3 and 5, both located in urban areas next to the stream's mouth, differ from each other due to the possible dilution of caffeine downstream of point 3 and domestic sewage discharge upstream of point 5.

In Utero Exposure to Caffeine and Acetaminophen, the Gut Microbiome, and Neurodevelopmental Outcomes: A Prospective Birth Cohort Study.

Pregnant individuals are exposed to acetaminophen and caffeine, but it is unknown how these exposures interact with the developing gut microbiome. We aimed to determine whether acetaminophen and/or caffeine relate to the childhood gut microbiome and whether features of the gut microbiome alter the relationship between acetaminophen/caffeine and neurodevelopment. Forty-nine and 85 participants provided meconium and stool samples at 6–7, respectively, for exposure and microbiome assessment. Fecal acetaminophen and caffeine concentrations were quantified, and fecal DNA underwent metagenomic sequencing. Caregivers and study staff assessed the participants’ motor and cognitive development using standardized scales. Prenatal exposures had stronger associations with the childhood microbiome than concurrent exposures. Prenatal acetaminophen exposure was associated with a trend of lower gut bacterial diversity in childhood [β = −0.17 Shannon Index, 95% CI: (−0.31, −0.04)] and was marginally associated with differences in the relative abundances of features of the gut microbiome at the phylum (Firmicutes, Actinobacteria) and gene pathway levels. Among the participants with a higher relative abundance of Proteobacteria, prenatal exposure to acetaminophen and caffeine was associated with lower scores on WISC-IV subscales. Acetaminophen during bacterial colonization of the naïve gut is associated with lasting alterations in childhood microbiome composition. Future studies may inform our understanding of downstream health effects.

Effects of Caffeine and 5-Caffeoylquinic Acid on Blood Cell In Vitro Cytokine Production in Response to Lipopolysaccharide Stimulation

Previous in vitro studies the have suggested anti-inflammatory properties of caffeine. Coffee is also rich in phenolic acids with several potential health benefits (chlorogenic acids), mainly 5-caffeoylquinic acid (5-CQA). The aim of the present study was to determine the effect of caffeine and 5-CQA, using concentrations comparable to typical human consumption, on LPS-induced cytokine production in whole blood in vitro cultures. Whole blood samples (n = 10) were stimulated for 24 h with LPS in the presence of caffeine or 5-CQA. The concentrations of various cytokines, malondialdehyde (MDA) and adenosine-3′,5′-cyclic monophosphate (cAMP) were determined in the culture media. Significant effects of caffeine were found only for the stimulated production of TNF-α (p = 0.014) and cAMP (p = 0.001). However, only the highest caffeine concentration (50 μM) induced lower TNF-α (p = 0.016) and higher cAMP (p = 0.006) levels in comparison to the control. The 5-CQA only influenced the stimulated production of IL-6 (p = 0.002), with lower values for the highest 5-CQA concentrations (2 μM: p = 0.012; 20 μM: p < 0.001). Neither caffeine (p = 0.227) nor 5-CQA (p = 0.259) influenced MDA concentrations. In conclusion, both caffeine and 5-CQA exerted limited in vitro anti-inflammatory effects. For caffeine, these effects were attained with high physiological concentrations. However, for 5-CQA, supraphysiological concentrations were needed.

Effects of Caffeine and COD from Coffee Wastewater on Anaerobic Ammonium Oxidation (Anammox) Activities

An anaerobic ammonium oxidation (anammox) process was employed to remove nitrogen from wastewater generated from a coffee brewing facility. The effects of caffeine and chemical oxygen demand (COD) in coffee wastewater on anammox activity were investigated. The anammox activity was inhibited in synthetic wastewater with a caffeine concentration greater than 350 mg/L. Daily additions of caffeine at 2.5 mg/L for 28 days to the same substrate did not inhibit anammox activity. However, daily additions of coffee wastewater with COD of ≥387 mg/L and caffeine at 2.5 mg/L significantly inhibited anammox activity. Because the pH was increased in the system, resulting in an increase in free ammonia (FA) concentration, one could postulate that FA is an inhibitor of anammox activity. Quantitative polymerase chain reaction (qPCR) analysis was employed to determine the populations of anammox and denitrifying bacteria. Coffee wastewater with bacterial COD to total nitrogen (bCOD:TN) ratios of 0.3–0.6:1 did not have any effect on the abundances of anammox and denitrifying bacteria. The results from this work suggest that biodegradable COD (bCOD) rather than total COD (TCOD) should be used for calculating the COD:TN ratio during the study of the effects of nitrogen removal from real wastewaters using the anammox process. A not-competitive model could fit the anammox inhibition with caffeine concentrations at 50–500 mg/L with maximum specific anammox activity (SAAmax) of 0.594 mg-N/mg-volatile suspended solids (VSS)/d and inhibitory constant (Ki) of 480.97 mg/L.

Effect of the dried limes (Citrus aurantifolia) on the caffeine kinetic extraction in black tea leaves (kinetic and thermodynamic study)

ABSTRACT. The kinetic and thermodynamic parameters were studied for the extraction of caffeine in black tea leaves and tea leaves containing antioxidant dried limes (Citrus aurantifolia). Caffeine is widely used in human consumption and is present in any liquid suitable for drinking except water. However, the health and environmental demerits make it significant to reduce caffeine to much less concentration or decaffeination. Therefore, the effect of the dried antioxidant Citrus aurantifolia on caffeine concentration followed by pseudo-first-order reaction, kinetic parameters (pseudo-first-order rate constant (k), equilibrium absorbance (A∞)), and thermodynamic activation parameters, the activation energy (Ea), change in enthalpy of activation (ΔH#), the change in entropy of activation (ΔS#) and change in activation Gibbs free energy (ΔG#) was calculated at four different temperatures 30, 60, 80, and 100 °C. Using the spectroscopic method the activation energy of the caffeine concentration in free black tea leaves solution (Ea = 13.32 kJmol-1) was higher than the black tea leaves mixed with lime solution (Ea = 9.89 kJmol-1), the higher collision of molecules A-factor (1.96 min-1), and large correlation factor (R2 = 0.95). The thermodynamic activation parameters ΔH#, ΔS#, and ΔG# demonstrate less heat absorption, irreversible, and more spontaneous interaction of lime with caffeine.
 
 KEY WORDS: Caffeine, Tea leaves, Citrus aurantifolia, Thermodynamic parameters, Kinetic
 Bull. Chem. Soc. Ethiop. 2022, 36(3), 697-705. 
 DOI: https://dx.doi.org/10.4314/bcse.v36i3.18

New Life of an Old Drug: Caffeine as a Modulator of Antibacterial Activity of Commonly Used Antibiotics.

With the rapid and continuous emergence of antimicrobial resistance, bacterial infections became a significant global healthcare concern. One of the proposed strategies to combat multidrug-resistant pathogens is to use additional compounds, such as natural biologically active substances, as adjuvants for existing antibiotics. In this study, we investigated the potential of caffeine, the widely consumed alkaloid, to modulate the antibacterial effects of antibiotics commonly used in clinical practice. We used disc diffusion assay to evaluate the effects of caffeine on 40 antibiotics in two Staphylococcus aureus strains (methicillin-resistant and methicillin-sensitive). Based on the results of this step, we selected five antibiotics for which the greatest caffeine-induced improvements in antibacterial activity were observed, and further analyzed their interactions with caffeine using a checkerboard approach. Caffeine at concentrations of 250 µg/mL or higher halved the MIC values of ticarcillin, cefepime, gentamycin, azithromycin, and novobiocin for all gram-negative species investigated (Pseudomonas aeruginosa, Klebsiella pneumoniae, and Acinetobacter baumannii). At the highest caffeine concentrations tested (up to 16 mg/mL), decreases in MIC values were 8- to 16-fold. The obtained results prove that caffeine modulates the activity of structurally diverse antibiotics, with the most promising synergistic effects observed for cefepime and azithromycin toward gram-negative pathogens.

Predictive factors of the contracture test for diagnosing malignant hyperthermia in a Brazilian population sample: a retrospective observational study

IntroductionMalignant Hyperthermia (MH) is a pharmacogenetic, hereditary and autosomal dominant syndrome triggered by halogenates/succinylcholine. The In Vitro Contracture Test (IVCT) is the gold standard diagnostic test for MH, and it evaluates abnormal skeletal muscle reactions of susceptible individuals (earlier/greater contracture) when exposed to caffeine/halothane. MH susceptibility episodes and IVCT seem to be related to individual features. ObjectiveTo assess variables that correlate with IVCT in Brazilian patients referred for MH investigation due to a history of personal/family MH. MethodsWe examined IVCTs of 80 patients investigated for MH between 2004‒2019. We recorded clinical data (age, sex, presence of muscle weakness or myopathy with muscle biopsy showing cores, genetic evaluation, IVCT result) and IVCT features (initial and final maximum contraction, caffeine/halothane concentration triggering contracture of 0.2g, contracture at caffeine concentration of 2 and 32 mmoL and at 2% halothane, and contraction after 100 Hz stimulation). ResultsMean age of the sample was 35±13.3 years, and most of the subjects were female (n=43 or 54%) and MH susceptible (60%). Of the 20 subjects undergoing genetic investigation, 65% showed variants in RYR1/CACNA1S genes. We found no difference between the positive and negative IVCT groups regarding age, sex, number of probands, presence of muscle weakness or myopathy with muscle biopsy showing cores. Regression analysis revealed that the best predictors of positive IVCT were male sex (+12%), absence of muscle weakness (+20%), and personal MH background (+17%). ConclusionsPositive IVCT results have been correlated to male probands, in accordance with early publications. Furthermore, normal muscle strength has been confirmed as a significant predictor of positive IVCT while investigating suspected MH cases.

Effect of low concentrations of caffeine and colchicine on microbial growth and biofilm formation

A study into the effect of low concentrations of caffeine and colchicine alkaloids on the growth and biofilm formation of rhizospheric microorganisms – Gram-positive Rhodococcus qingshengii and Gramnegative Rhizobium radiobacter – is presented. Alkaloids present in plants effectively protect them from being eaten by animals of various taxa, as well as from fungal and bacterial infections. Forming part of a complex, evolutionary system of plant-microbial interactions, they occur naturally in the medium at low concentrations. One of the decisive factors in the spread of microorganisms is the formation of biofilms. In the study, alkaloids were added to the bacterial suspension at time intervals corresponding to distinct phases of biofilm formation. In order to determine the level of biofilm formation, bacterial cells were stained with crystal violet. The optical density of the suspension was used to assess the effect of alkaloids on bacterial growth and biofilm formation. The effect of caffeine and colchicine in concentrations of 10-5 g/L on the growth and biofilm formation of microorganisms is revealed for the first time. Although alkaloids did not affect the growth of the suspension of Gram-positive Rhodococcus qingshengii, they effectively suppressed the suspension of Gram-negative Rhizobium radiobacter. While Rhodococcus qingshengii showed greater sensitivity to alkaloids at the stage of biofilm formation, Rhizobium radiobacter was more sensitive at the stage of "mature" biofilms. The effect of alkaloids could be expressed both in terms of suppression and enhancement of biofilm growth. It can be assumed that the mechanism of action of plant alkaloids at the level of signaling systems and bacterial quorum sensing allows the protective function to be maintained even with a significant decrease in the number of alkaloids under conditions of permanent antagonism in which they are constantly being destroyed by phytopathogens.

Distribution and Ecological Risk Assessment of Pharmaceuticals and Personal Care Products in Sediments of North Canal, China

The pollution of water bodies by pharmaceuticals and personal care products (PPCPs) has attracted widespread concern due to their widespread use and pseudo-persistence, but their effects on sediments are less known. In this study, solid-phase extraction-high performance liquid chromatography–tandem mass spectrometry (SPE-LC/MSMS) was used to investigate the occurrence and ecological risks of five typical pharmaceuticals and personal care products (PPCPs) in thirteen key reservoirs, sluices, dams, and estuaries in the Haihe River Basin. At the same time, the PPCP exchanges of surface water, groundwater, and sediments in three typical sections were studied. Finally, the PPCP’s environmental risk is evaluated through the environmental risk quotient. The results showed that the five PPCPs were tri-methoprazine (TMP), sinolamine (SMX), ibuprofen (IBU), triclosan (TCS), and caffeine (CAF). The average concentration of these PPCPs ranged from 0 to 481.19 μg/kg, with relatively high concentrations of TCS and CAF. The relationship between PPCPs in the surface sediments was analyzed to reveal correlations between SMX and TMP, CAF and IBU, CAF and TCS. The risk quotients (RQ) method was used to evaluate the ecological risk of the five detected PPCPs. The major contributors of potential environmental risks were IBU, TCS and CAF, among which all the potential environmental risks at the TCS samples were high risk. This study supplemented the research on the ecological risk of PPCPs in sediments of important reaches of the North Canal to reveal the importance of PPCP control in the North Canal and provided a scientific basis for pollution control and risk prevention of PPCPs.

The effects of caffeine on the motility and viability of stallion spermatozoa at different temperature conditions.

The purpose of this study was to evaluate the dose- and time-dependent effect of caffeine treatment on the motility and viability of stallion spermatozoa at different temperatures. Six dose groups (A to F) were established with changing caffeine concentrations (from 0.625 to 10 mg/mL). The control samples were prepared by diluting the ejaculate only with physiological salt solution. The samples were examined after 0, 1, 2 and 3 h of incubation at 5 °C and 37 °C. The motility parameters were evaluated by Computer Assisted Semen Analyzer (CASA) system, and the viability was assessed by the mitochondrial toxicity test at the end of the incubation. A positive effect of the lowest tested caffeine concentration on the motility parameters was observed throughout the incubation period at 5 °C. At the end of the 3h incubation, the viability in every sample in these groups, treated with any caffeine concentration, showed lower values compared to the control. At the higher incubation temperature (37 °C), caffeine positively affected the motility in samples B (P < 0.05) and D, E, F (P < 0.001) after 3 h of incubation; however, the viability showed a slightly decreasing tendency. Our results suggest that caffeine, in an optimal concentration, may be used as a component of stallion semen extenders.

Impact of origin environment to the chemical characteristic of java coffees

Coffee is one of the most important export commodities and a unique product which have different taste, shape and flavour according to the origin. The objective of this research was to characterized three different origins of Java coffee (Java Preanger, Andung Sari and Malang) using chemical method. Determination of major composition of coffees were carried out using proximate analysis and determination of caffeine, trigonelline and chlorogenic acid (CGA) was conducted using LCMS method. The concentration of each chemical composition were significantly different among three Java coffees, except for trigonelline and lipid concentration. The carbohydrate and protein concentration of Andung Sari coffee were the highest beside another two coffee origins which were 58.4% and 14.59% respectively. The highest concentration of CGA was obtained in Andung Sari coffee (9.1%) followed by Malang coffee (8.32%) and Java Preanger coffee (6.04%). The highest concentration of caffeine was also obtained in Andung Sari (1.42%). The Chemical quality of Andung Sari coffee is superior among Java coffees.