Toxicity of cylindrospermopsin in human lymphocytes: Proliferation,viability and cell cycle studies

The global expansion of cylindrospermopsin (CYN) producing cyanobacteria in surface freshwater increases the risk of human exposure and poisoning. Following ingestion, CYN is transported with blood in general circulation to the liver and kidneys, and can potentially interact with immune system cells. In the present study, we investigated whether CYN (0.01–1.0 μg ml⁻¹) can alter the function of human peripheral blood lymphocytes isolated from healthy donors. It was found that CYN demonstrates significant antiproliferative activity in lymphocytes during different phases of their activation. The most remarkable effects (decrease by > 90%) were observed in lymphocytes exposed to 1 μg ml⁻¹ CYN at the beginning of activation. Further analyses revealed a cell-cycle arrest at G0/G1 and prolonged S phase in lymphocytes undergoing activation and significant apoptosis inducement in activated cells. Reduced abilities to fight pathogenic microorganisms or malignant cells should be taken into consideration in CYN exposure and risk assessments.     

Toxic effects induced by mycotoxin fumonisin B1 on equine spermatozoa: Assessment of viability,sperm chromatin structure stability,ROS production and motility

Fumonisin B1 (FB1) is a mycotoxin produced by Fusarium species that exerts its toxic effect through interference with the sphingolipid pathway by inhibiting ceramide synthase. A FB1-dependent sperm toxicity was reported in boars. No information on FB1-related reproductive toxicity in stallions, the most sensitive animal species, has been reported. The aim of the present study was to assess the in vitro toxicity of FB1 on fresh and frozen-thawed equine spermatozoa by analyzing sperm viability, chromatin stability (SCSA) and reactive oxygen species (ROS) production by flow cytometry and sperm motility by CASA system. Fumonisin B1 did not affect viability of fresh spermatozoa after 2 h exposure up to 25 μM. Damage on sperm chromatin structure was observed only in one frozen sample after exposure up to 2.5 × 10⁻⁵ μM FB1 without associated increase of ROS. Increase of ROS, at FB1 levels up to 2.5 × 10⁻⁴ μM, was found on another frozen-thawed sperm sample, may be as a consequence of seminal plasma removal. At 7.5 and 15 μM, FB1 induced reduction of total and progressive motility.     

Emodin inhibits human sperm functions by reducing sperm [Ca2+]i and tyrosine phosphorylation

Emodin, a bioactive anthraquinone widely used in Chinese traditional medicine, disrupts mouse testicular gene expression in vivo. In this study, we investigated the toxicity of emodin to human sperm in vitro. Different doses of emodin (25, 50, 100, 200 and 400 μM) were applied to ejaculated human sperm. The results indicated that 100, 200 and 400 μM emodin significantly inhibited the total motility, progressive motility and linear velocity of human sperm. In addition, sperm's ability to penetrate viscous medium together with progesterone induced capacitation and acrosome reaction was also adversely affected by emodin. In contrast, emodin did not affect sperm viability. Furthermore, intracellular Ca²⁺ concentration ([Ca²⁺]_i) and tyrosine phosphorylation, which serve as key regulators of sperm function, were dose-dependently reduced by emodin (50–400 μM). These results suggest that emodin inhibits human sperm functions by reducing sperm [Ca²⁺]_i and suppressing tyrosine phosphorylation in vitro.     

Influence of experimentally induced fever on the disposition of cefpirome in buffalo calves

The influence of Escherichia coli endotoxin-induced fever on the disposition of cefpirome was investigated in five male buffalo calves following a single intravenous dose of 10 mg kg⁻¹. Blood samples were collected from 1 min to 24 h of drug administration. The drug concentration in plasma was estimated by microbiological assay using E. coli as a test organism. The disposition of cefpirome followed two-compartment open model and the drug was detected above the minimum inhibitory concentration in plasma up to 12 h. The Vd_area and AUC were 0.75 ± 0.01 L kg⁻¹ and 35.1 ± 0.46 μg ml⁻¹ h, respectively. The elimination half-life of 1.81 ± 0.009 h and Cl_B of 0.29 ± 0.004 L kg⁻¹ h⁻¹ reflected rapid elimination and body clearance of cefpirome in febrile buffalo calves. Based on the results, a satisfactory dosage regimen of cefpirome in febrile buffalo calves was calculated to be 6 mg kg⁻¹ to be repeated at 8 h intervals.     

Industrial release of fluoroquinolones (FQs) in the waste water bodies with their associated ecological risk in Pakistan

The unchecked production and use of fluoroquinolones (FQs) for the treatment of infections in human and livestock has increased in Pakistan, which resulted in large amount of antibiotics in water bodies. In the current study, the prevalence and associated ecological risk of three FQs were investigated in waste-water bodies and sludge samples of Kahuta and Hattar industrial zones. The average concentrations of ciprofloxacin (CIP), enrofloxacin (ENR) and levofloxacin (LEV) in the waste-water samples were slightly higher in Kahuta (i.e. 58, 32.9, and 36.7 μg L⁻¹ respectively), than those in Hattar sites (i.e. 42.1, 41.2, and 48.9 μg L⁻¹ respectively). However, the concentrations of CIP, ENR and LEV in the sludge samples were significantly higher (i.e. 159; 153 and 164 μg kg⁻¹ respectively) in Hattar sites, compared to those in Kahuta sites (i.e. 129, 58 and 91 μg kg⁻¹ respectively). The uses of FQs in the health sector resulted in water pollution and poses the ecological risk to aquatic organisms. The individual risk associated with CIP was highest in Kahuta industrial sites for green algae ranging (2900–9100) followed by M. aeruginosa (5800–18200), cyanobacteria (580–18204) and invertebrates (24.2–75.8). These values suggested that the prevalence of antibiotics in the waste-disposal sites could be potential risk for the aquatic ecosystem, and harmful to biodiversity.     

Oxidative stress responses in zebrafish Danio rerio after subchronic exposure to atrazine

Atrazine is one of the most used pesticides all over the world and it is frequently detected in surface water. The aim of this study was to investigate if zebrafish exposure to atrazine could induce oxidative stress and changes in detoxifying system. Juvenile fish were exposed to sublethal concentrations of 0.3, 3, 30, or 90 μg L⁻¹ for 28 days. The level of oxidized lipids increased in experimental groups exposed to atrazine at 30 and 90 μg L⁻¹ compared to control. Activity of glutathione S-transferase decreased in group with the highest concentration compared to control. A significant decline was observed in catalase activity in all experimental groups compared to control. Activity of superoxide dismutase increased only in experimental group exposed to atrazine at 30 μg L⁻¹ compared to control. Activity of glutathione peroxidase and reductase (GR) increased in experimental groups exposed to atrazine at 0.3 (only for GR activity) and 90 μg L⁻¹ compared to control. Our results showed that atrazine exposure had profound influence on the oxidative stress markers and detoxifying enzyme of the exposed zebrafish. The changes in antioxidant enzyme activities could be an adaptive response to protect the fish from the atrazine-induced toxicity.     

Boar spermatozoa as a biosensor for detecting toxic substances in indoor dust and aerosols

The presence, quantity and origins of potentially toxic airborne substances were searched in moisture damaged indoor environments, where building related ill health symptoms were suspected and reference sites with no health complaints. Boar spermatozoa were used as the toxicity sensor. Indoor aerosols and dusts were collected from kindergartens, schools, offices and residences (n = 25) by electrostatic filtering, vacuuming, wiping from elevated surfaces and from the interior of personal computers. Toxicity was measured from the ethanol or methanol extracts of the dusts and aerosols. EC₅₀ was expressed as the lowest concentration of the airborne substance that inhibited motility of >50% of the exposed sperm cells compared to vehicle control, within 30 min, 1 day or 3–4 days of exposure. Remarkably toxic aerosols (EC₅₀ ⩽6 μg ml⁻¹) were found from 11 sites, all of these were sites with known or suspected for building related ill health. Toxic microbial cultures were obtained from subsamples of the toxic aerosols/dusts. From these cereulide, amylosin, valinomycin and a novel indoor toxin, stephacidin B were identified and toxicities measured. Airborn dispersal of valinomycin from Streptomyces griseus cultures was evaluated using a flow-through chamber. Significant amounts of valinomycin (LC–MS assay) and toxicity (boar sperm motility assay) were carried by air and were after 14 days mainly recovered from the interior surfaces of the flow chamber.     

Considerations using silver nitrate as a reference for in vitro tests with silver nanoparticles

Most in vitro tests regarding the cellular toxicology of nanoparticulate metals compare particle to associated metal ion exposure. However, it is also a fact, that for example silver ions are reduced by sugars or transformed to silver chloride by chloride salts which are abundant components of cell culture media. These reactions are likely to either complicate or even invalidate comparisons between effects of ions and particles. Here, we present a fast and quantitative method to determine particle formation and numbers in different cell culture media with non-destructive small-angle X-ray scattering (SAXS). Silver nitrate with a concentration of 25 μg Ag mL^− 1 was dissolved for up to 24 h at 37 °C in Dulbeccos Modified Eagle Medium (DMEM) with and without 10% fetal bovine serum (FBS) and a solution of D-glucose (4.5 μg mL^− 1), respectively. Silver nanoparticles were observed in all solutions after 5 min. The cell culture media displayed a limited particle-growth. FBS showed an effect on the polydispersity of the generated particles but after 5 min the overall particle size was nearly equal in FBS and non FBS supplemented medium. Particles in D-glucose were precipitating after 10 min. Particulate silver concentration was between 3 and 4 μg mL^− 1 in both cell culture media (CCM). These results should be taken into account when performing silver ion-toxicity experiments in relevant media.     

In vivo effects of deoxynivalenol (DON) on innate immune responses of carp (Cyprinus carpio L.)

Deoxynivalenol (DON) is one of the most important members of Fusarium toxins since it often can be found in relevant concentrations in animal feeds. The effects of this group of toxins on fish are mostly unknown. The present study shows results from a feeding trial with carp (Cyprinus carpio L.) using three different concentrations of DON (352 μg kg⁻¹, 619 μg kg⁻¹, and 953 μg kg⁻¹ final feed, respectively) which are comparable to levels found in commercial fish feeds. Effects on growth and mass of fish were not observed during this 6 weeks lasting experiment. Only marginal DON concentrations were found in muscle and plasma samples. Blood parameters were not influenced although smaller erythrocytes occurred in fish treated with 352 μg kg⁻¹ DON. Analysis of antioxidative enzymes in erythrocytes showed increased superoxid dismutase and catalase activities in fish fed the low-dose feed. Immunosuppressive effects of DON were confirmed whereby cytotoxic effects on immune cells only partly explained the impairment of innate immune responses. Exact polarization of the immune system into pro-inflammatory or anti-inflammatory responses due to DON exposure should be clarified in further experiments, especially since the current results raise concern about impaired immune function in fish raised in aquaculture.     

Urinary analysis reveals high deoxynivalenol exposure in pregnant women from Croatia

In this pilot survey the levels of various mycotoxin biomarkers were determined in third trimester pregnant women from eastern Croatia. First void urine samples were collected and analysed using a “dilute and shoot” LC–ESI–MS/MS multi biomarker method. Deoxynivalenol (DON) and its metabolites: deoxynivalenol-15-glucuronide and deoxynivalenol-3-glucuronide were detected in 97.5% of the studied samples, partly at exceptionally high levels, while ochratoxin A was found in 10% of the samples. DON exposure was primarily reflected by the presence of deoxynivalenol-15-glucuronide with a mean concentration of 120 μg L⁻¹, while free DON was detected with a mean concentration of 18.3 μg L⁻¹. Several highly contaminated urine samples contained a third DON conjugate, tentatively identified as deoxynivalenol-7-glucuronide by MS/MS scans. The levels of urinary DON and its metabolites measured in this study are the highest ever reported, and 48% of subjects were estimated to exceed the provisional maximum tolerable daily intake (1 μg kg⁻¹ b.w.).     

Ecotoxicity of veterinary enrofloxacin and ciprofloxacin antibiotics on anuran amphibian larvae

The ecological risks posed by two β-diketone antibiotics (DKAs, enrofloxacin, ENR and ciprofloxacin, CPX), characterized by their long persistence in aqueous environments and known deleterious effect on model organisms such as zebrafish were analysed using Rhinella arenarum larvae. Sublethal tests were conducted using environmentally relevant concentrations of both ENR and CPX (1–1000 μg L⁻¹) under standard laboratory conditions for 96 h. Biological endpoints and biomarkers evaluated were body size, shape, development and growth rates, and antioxidant enzymes (glutathione-S-transferase, GST; Catalase, CAT). Risk assessment was analysed based on ration quotients (RQ). The size and shape measurements of the larvae exposed to concentrations greater than 10 μg L⁻¹ of CPX were lower compared to controls (Dunnett post hoc p < 0.05) and presented signs of emaciation. Concentrations of 1000 μg L⁻¹of CPX induced GST activity, in contrast with inhibited GST and CAT of larvae exposed to ENR. Risk assessments indicated that concentrations greater than or equal to10 μg L⁻¹ of CPX and ENR are ecotoxic for development, growth, detoxifying, and oxidative stress enzymes. It is suggested that additional risk assessments may provide evidence of bioaccumulation of CPX and ENR in tissues or organs of amphibian larvae by mesocosm sediment test conditions. Finally, intestinal microbiome studies should be considered to establish the mechanisms of action of both antibiotics.     

Chemical state of heterocyclic aromatic amines in grilled beef: Evaluation by in vitro digestion model and comparison of alkaline hydrolysis and organic solvent for extraction

During grilling of the roast beef the following heterocyclic aromatic amines were found: IQ = 200.6 ng 100 g⁻¹, MeIQx = 719.8 ng 100 g⁻¹, MeIQ = 532.9 ng 100 g⁻¹, 4.8-diMeIQx = 755.4 ng 100 g⁻¹, norharmane = 507.0 ng 100 g⁻¹, harmane = 1952.6 ng 100 g⁻¹, Phe-P 1 = 263.7 ng 100 g⁻¹, Trp-P 2 = 559.2 ng 100 g⁻¹, PhIP = 1179.8 ng 100 g⁻¹ and AαC = 51.7 ng 100 g⁻¹. Their content was tested by using the method based on alkaline hydrolysis of the sample and the method based on solvent extraction of the grilled meat samples at different temperatures (without hydrolysis). The study showed that the heterocyclic aromatic amines produced during the grilling of beef are in a free form and chemically or physico-chemically bonded. The chemical forms of HAA formed in food have never been studied. For the purpose of the partial confirmation that HAA may be chemically or physico-chemically bonded, grilled beef samples were digested in vitro in model segments of the human digestive tract. Digestive enzymes, particularly proteolytic enzymes caused a statistically significant increase of free HAA determined by using solvent extraction without prior chemical hydrolysis of the sample.     

Preparation of estradiol chitosan nanoparticles for improving nasal absorption and brain targeting

The estradiol(E₂)-loaded chitosan nanoparticles (CS-NPs) were prepared by ionic gelation of chitosan with tripolyphosphate anions (TPP). The CS-NPs had a mean size of (269.3 ± 31.6) nm, a zeta potential of +25.4 mV, and loading capacity of E₂ CS-NPs suspension was 1.9 mg ml⁻¹, entrapment efficiency was 64.7% on average. Subsequently, this paper investigated the levels of E₂ in blood and the cerebrospinal fluid (CSF) in rats following intranasal administration of E₂ CS-NPs. E₂-loaded CS-NPs were administered to male Wister rats either intranasally or intravenously at the dose of 0.48 mg kg⁻¹. The plasma levels achieved following intranasal administration (32.7 ± 10.1 ng ml⁻¹; t_max 28 ± 4.5 min) were significantly lower than those after intravenous administration (151.4 ± 28.2 ng ml⁻¹), while CSF concentrations achieved after intranasal administration (76.4 ± 14.0 ng ml⁻¹; t_max 28 ± 17.9 min) were significantly higher than those after intravenous administration (29.5 ± 7.4 ng ml⁻¹ t_max 60 min). The drug targeting index (DTI) of nasal route was 3.2, percent of drug targeting (DTP%) was 68.4%. These results showed that the E₂ must be directly transported from the nasal cavity into the CSF in rats. Finally, compared with E₂ inclusion complex, CS-NPs improved significantly E₂ being transported into central nervous system (CNS).     

Toxicological effects of clofibric acid and diclofenac on plasma thyroid hormones of an Indian major carp,Cirrhinus mrigala during short and long-term exposures

In the present investigation, the toxicity of most commonly detected pharmaceuticals in the aquatic environment namely clofibric acid (CA) and diclofenac (DCF) was investigated in an Indian major carp Cirrhinus mrigala. Fingerlings of C. mrigala were exposed to different concentrations (1, 10 and 100 μg L⁻¹) of CA and DCF for a period of 96 h (short term) and 35 days (long term). The toxic effects of CA and DCF on thyroid hormones (THs) such as thyroid stimulating hormone (TSH), thyroxine (T₄) and triiodothyronine (T₃) levels were evaluated. During the short and long-term exposure period TSH level was found to be decreased at all concentrations of CA (except at the end of 14^th day in 1 and 10 μg L^−l and 21^st day in 1 μg L^−l) whereas in DCF exposed fish TSH level was found to be increased when compared to control groups. T₄ level was found to be decreased at 1 and 100 μg L^−l of CA exposure at the end of 96 h. However, T₄ level was decreased at all concentrations of CA and DCF during long-term (35 days) exposure period. Fish exposed to all concentrations of CA and DCF had lower level of T₃ in both the treatments. These results suggest that both CA and DCF drugs induced significant changes (P < 0.01 and P < 0.05) on thyroid hormonal levels of C. mrigala. The alterations of these hormonal levels can be used as potential biomarkers in monitoring of pharmaceutical drugs in aquatic organisms.     

Activity of the enzymes of the antioxidative system in cadmium-treated Oxya chinensis (Orthoptera Acridoidae)

One purpose in this research was to determine the toxic effects of Cd on antioxidant enzymes of Oxya chinensis (Orthoptera: Acridoidae). Changes in the activities of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and guaiacol peroxidase (GPx) were measured in O. chinensis insects injected with Cd²⁺. Fifth-nymphs of O. chinensis insects were injected with Cd²⁺ at different concentrations (0, 0.55 × 10⁻⁴, 1.10 × 10⁻⁴, 1.65 × 10⁻⁴, 2.20 × 10⁻⁴, and 2.75 × 10⁻⁴ g g⁻¹). An increase in SOD activity in O. chinensis was observed at 1.10 × 10⁻⁴ to 2.75 × 10⁻⁴ g g⁻¹ Cd²⁺. The SOD activity was lower at 2.20 × 10⁻⁴and 2.75 × 10⁻⁴ g g⁻¹ than that at 1.10 × 10⁻⁴ and 1.65 × 10⁻⁴ g g⁻¹. It appears that SOD had a positive protective effect at low Cd²⁺ concentrations, and that this effect disappeared at high Cd²⁺ concentrations. CAT activity was accelerated to varying degrees at 1.10 × 10⁻⁴ to 2.75 × 10⁻⁴ g g⁻¹ for males and at 1.10 × 10⁻⁴, 2.20 × 10⁻⁴, and 2.75 × 10⁻⁴g g⁻¹ for females. CAT showed a strong detoxification effect with all treatments. GPx activity decreased with increasing Cd²⁺ concentration with all treatments for males and at 2.20 × 10⁻⁴ and 2.65 × 10⁻⁴g g⁻¹ for females. We showed that GPx activity had a weak detoxification function with all treatments for males and at high Cd²⁺ for females. Thus, CAT had a strong detoxification effect, whereas SOD had a medium and GPx had a weak detoxification effect. Among the three enzymes, CAT played an important role in the damaging mechanisms of reactive oxygen species in O. chinensis insects. Alterations of the antioxidant enzyme level under environmental stresses are suggested as indicators of biotic and abiotic stress.     

Evaluation of estrogenic activity in animal diets using in vitro assay

A yeast estrogen bioassay (RIKILT REA) was in-house validated for feed on the 5 μg 17β-estradiol-equivalents per kg level according to EC Decision 2002/657/EC. All the performance characteristics met the criteria as defined in the Decision and the REA is able to detect 17β-estradiol in animal feed at a low level of 1.15–2 μg kg⁻¹. Subsequently, the developed and validated procedure was applied to determine the estrogenic activity in 24 feed samples intended for food producing animals, pets and laboratory animals. Two batches of rodent diet Murigran and one dog feed have been presented as a suspect, i.e. gave responses above the determined decision limit (CCα) and detection capability (CCβ). In assessing the performance of the estrogenic activity in these diets evaluated by comparison with the 17β-estradiol calibration curve, 17β-estradiol-equivalence levels of 7.07 μg EEQ kg⁻¹ and 9.54 μg EEQ kg⁻¹ in two batches of rodent diet and 5.3 μg EEQ kg⁻¹ in dog feed have been established. The activities observed in the rodent feed could be explained by chemical analysis, revealing high amounts of genistein, daidzein and trace amounts of zearalenone. In addition, the estrogenic activity in one of rodent feed was above the established CCα, but below the CCβ values established and all other samples showed no estrogenic activity with responses below the CCα value, which corresponds to levels below 2 μg EEQ kg⁻¹.     

Human health risk assessment and dietary intake of organochlorine pesticides through air,soil and food crops (wheat and rice) along two tributaries of river Chenab,Pakistan

To assess the organochlorine pesticides (OCPs) contamination and their probable hazardous effects on human health; cereal crops (wheat and rice; n = 28) agricultural soil (n = 28) and air (n = 6) samples were collected from Gujranwala division, Punjab Province, Pakistan. ∑OCPs concentration ranged between 123 and 635 pg m⁻³, 31 and 365 ng g⁻¹ (dw), 2.72 and 36.6 ng g⁻¹ (dw), 0.55 and 15.2 ng g⁻¹ (dw) for air, soil, rice and wheat samples, respectively. DDTs were the predominant over other OCPSs detected from screened samples while the source apportionment analysis suggested the new inputs of DDTs in the study area. EDI (estimated daily intake) of ∑OCPs through rice and wheat was found 39 and 40 ng kg⁻¹ day⁻¹, respectively. Hazard ratios (HRs) on the basis 95th percentile concentrations were exceeding the integrity for most of the investigated OCP in rice and wheat. The results revealed that there is a severe risk to the human population of the study area through consumption of contaminated cereal crops.     

LC–MS/MS determination of the isomeric neurotoxins BMAA (β-N-methylamino-l-alanine) and DAB (2,4-diaminobutyric acid) in cyanobacteria and seeds of Cycas revoluta and Lathyrus latifolius

Since diverse taxa of cyanobacteria has been linked to biosynthesis of BMAA, a controversy has arisen about the detection of neurotoxic amino acids in cyanobacteria. In this context, a novel LC–MS/MS method was developed for the unambiguous determination of β-N-methylamino-l-alanine (BMAA) and 2,4-diaminobutyric acid (DAB) in cyanobacteria and selected plant seeds. Both neurotoxic and non-proteinogenic amino acids were analyzed without derivatization considering the total concentration of the free and protein-bound form. The investigation of overall 62 cyanobacterial samples of worldwide origin by application of this method revealed the absence of BMAA, whereas seeds of Cycas revoluta contained 6.96 μg g^?1 of free BMAA. In contrast, the isomer DAB was confirmed in 16 cyanobacterial samples in concentrations of 0.07–0.83 μg g^?1,whereof one sample is distributed as nutritional supplement. In addition, seeds of Lathyrus latifolius contained 4.21 μg g^?1 of free DAB. Limits of detection were for BMAA < 1.0 μg g^?1 in the cyanobacterial matrix and < 0.14 μg g^?1 in angiosperm seeds. DAB exhibits higher sensitivities of <0.06 μg g^?1 in cyanobacteria and <0.008 μg g^?1 in angiosperm seeds. The highly specific analysis method with increased detection sensitivity eliminates the disadvantages of derivatization-based methods to be discussed.     

Effects of nonylphenol on motility and subcellular elements of epididymal rat sperm

Nonylphenol (NP) is an important environmental toxicant and potential endocrine disrupting chemical. The objective of these studies was to determine the effects of NP on epididymal rat sperm in vitro. Epididymal sperm samples from Sprague-Dawley rats were incubated in 1, 10, 100, 250, and 500 μg/ml NP for 1, 2, 3, or 4 h. Computer-assisted sperm analysis was used to determine motility. Epifluorescent microscopy was used to determine acrosomal status and flow cytometry was used to determine mitochondrial membrane potential (MMP) and chromatin integrity. Exposure of epididymal rat sperm to 250 or 500 μg/ml NP was highly detrimental to motility (P < 0.05), with complete loss of motility observed after exposure to 500 μg/ml NP (P < 0.05). The acrosomal integrity of sperm was significantly reduced with the lowest concentration (1 μg/ml) of NP, and higher concentrations resulted in a dose-dependent induction of the acrosomal reaction (P < 0.05). Similarly, the percentage of sperm with high MMP declined dramatically after exposure to 100, 250, and 500 μg/ml NP (P < 0.05). Duration of NP exposure did not have any effect on motility or MMP and NP did not appear to have detrimental effects on chromatin integrity (P > 0.05). These results indicate that major mechanism of action of NP on rat sperm is by adversely affecting their acrosomal integrity. However, NP-induced impaired sperm motility, decreased mitochondrial membrane potential also likely to play an important role in destruction of sperm function.     

Efficient Ibuprofen Delivery from Anhydrous Semisolid Formulation Based on a Novel Cross-linked Silicone Polymer Network: An In Vitro and In Vivo Study

The use of silicone as a primary polymer in topical semisolid pharmaceutical formulations is infrequent. Recent development of novel silicone materials provides an opportunity to investigate their drug delivery efficiencies. In this study, an anhydrous semisolid formulation was prepared using a novel cross-linked silicone polymer network swollen in isododecane. Similar formulations were prepared using petrolatum, an acrylic, or a cellulose polymer. All formulations contained 5% ibuprofen (IBP). In vitro permeability was evaluated for all formulations and a commercial product using human cadaver epidermis. The silicone formulation delivered IBP more efficiently than all other formulations in terms of flux, cumulative amount, and percent drug release. The silicone formulation showed the maximum flux of 85.9 μg.cm⁻².h⁻¹ and a cumulative IBP release of 261.6 μg in 8 h, whereas the benchmark showed 20.1 μg.cm⁻².h⁻¹ and 30.9 μg, respectively. An in vivo study conducted on rats showed calculated blood AUCs of 59.2 and 17.6 μg.h/g (p < 0.003) for the silicone formulation and the benchmark, respectively. The IBP in excised rat skin was 264 ± 59 μg/g for the silicone formulation and 102 ± 5 μg/g for the benchmark. The results obtained from the in vitro and in vivo studies demonstrate efficient topical IBP delivery by the silicone formulation.