Qualification of a non-animal vaginal irritation method admitted as nonclinical assessment model (NAM) in the Incubator Phase of the United States Food and Drug Administration (US FDA) Medical Devices Development Tool (MDDT).

The U.S. Food and Drug Administration (FDA) Center for Devices and Radiological Health (CDRH) classifies personal lubricants as Class II medical devices. Because of this status and the nature of body contact common to personal lubricants, CDRH reviewers routinely recommend a standard biocompatibility testing battery that includes: an in vivo rabbit vaginal irritation (RVI) test; an in vivo skin sensitization test, such as the guinea pig maximization test (GPMT); and an in vivo acute systemic toxicity test using mice or rabbits. These tests are conducted using live animals, despite the availability of in vitro and other non-animal test methods that may be suitable replacements. The only test included in the biocompatibility battery currently conducted using in vitro assay(s) is cytotoxicity. FDA's recently launched Predictive Toxicology Roadmap calls for the optimization of non-animal methods for the safety evaluation of drugs, consumer products and medical devices. In line with these goals, a Consortium comprising the Institute for In Vitro Sciences, Inc. (IIVS), industry, the Consumer Healthcare Products Association (CHPA), and the PETA International Science Consortium (PETA-ISC) is qualifying the use of an in vitro testing method as replacement for the RVI test. Participating companies include manufacturers of personal lubricants and those interested in the advancement of non-animal approaches working collaboratively with the FDA CDRH to develop an in vitro testing approach that could be used in place of the RVI in pre-market submissions. Personal lubricants and vaginal moisturizers with diverse chemical and physical properties (e.g., formulation, viscosity, pH, and osmolality) in their final undiluted form will be the focus of the program. In vitro vaginal irritation data generated using commercially available human reconstructed vaginal tissue model(s) will be paired with existing in vivo RVI data and analyzed to develop a Prediction Model for the safety assessment of these products. This research plan has been accepted into the FDA CDRH Medical Device Development Tools (MDDT) program as a potential non-clinical assessment model (NAM). The proposed NAM aligns with the goals of the recently launched FDA Roadmap to integrate predictive toxicology methods into safety and risk assessment with the potential to replace or reduce the use of animal testing.

Ornamental plants on sale to the public are a significant source of pesticide residues with implications for the health of pollinating insects.

Garden centres frequently market nectar- and pollen-rich ornamental plants as "pollinator-friendly", however these plants are often treated with pesticides during their production. There is little information on the nature of pesticide residues present at the point of purchase and whether these plants may actually pose a threat to, rather than benefit, the health of pollinating insects. Using mass spectrometry analyses, this study screened leaves from 29 different 'bee-friendly' plants for 8 insecticides and 16 fungicides commonly used in ornamental production. Only two plants (a Narcissus and a Salvia variety) did not contain any pesticide and 23 plants contained more than one pesticide, with some species containing mixtures of 7 (Ageratum houstonianum) and 10 (Erica carnea) different agrochemicals. Neonicotinoid insecticides were detected in more than 70% of the analysed plants, and chlorpyrifos and pyrethroid insecticides were found in 10% and 7% of plants respectively. Boscalid, spiroxamine and DMI-fungicides were detected in 40% of plants. Pollen samples collected from 18 different plants contained a total of 13 different pesticides. Systemic compounds were detected in pollen samples at similar concentrations to those in leaves. However, some contact (chlorpyrifos) and localised penetrant pesticides (iprodione, pyroclastrobin and prochloraz) were also detected in pollen, likely arising from direct contamination during spraying. The neonicotinoids thiamethoxam, clothianidin and imidacloprid and the organophosphate chlorpyrifos were present in pollen at concentrations between 6.9 and 81 ng/g and at levels that overlap with those known to cause harm to bees. The net effect on pollinators of buying plants that are a rich source of forage for them but simultaneously risk exposing them to a cocktail of pesticides is not clear. Gardeners who wish to gain the benefits without the risks should seek uncontaminated plants by growing their own from seed, plant-swapping or by buying plants from an organic nursery.

Evaluation of the importance of astrocytes when screening for acute toxicity in neuronal cell systems.

Reliable, high throughput, in vitro preliminary screening batteries have the potential to greatly accelerate the rate at which regulatory neurotoxicity data is generated. This study evaluated the importance of astrocytes when predicting acute toxic potential using a neuronal screening battery of pure neuronal (NT2.N) and astrocytic (NT2.A) and integrated neuronal/astrocytic (NT2.N/A) cell systems derived from the human NT2.D1 cell line, using biochemical endpoints (mitochondrial membrane potential (MMP) depolarisation and ATP and GSH depletion). Following exposure for 72 h, the known acute human neurotoxicants trimethyltin-chloride, chloroquine and 6-hydroxydopamine were frequently capable of disrupting biochemical processes in all of the cell systems at non-cytotoxic concentrations. Astrocytes provide key metabolic and protective support to neurons during toxic challenge in vivo and generally the astrocyte containing cell systems showed increased tolerance to toxicant insult compared with the NT2.N mono-culture in vitro. Whilst there was no consistent relationship between MMP, ATP and GSH log IC(50) values for the NT2.N/A and NT2.A cell systems, these data did provide preliminary evidence of modulation of the acute neuronal toxic response by astrocytes. In conclusion, the suitability of NT2 neurons and astrocytes as cell systems for acute toxicity screening deserves further investigation.

Using the family ecosystem model to enhance pastoral care and counseling.

Argues that with individual, family, and world issues becoming increasingly complicated, those involved in pastoral care can benefit from using a comprehensive ecological approach to understanding and helping those in need. Suggests that the "family ecosystem model" that emphasizes interactions among individuals/families and their multifaceted environments, including their natural, human-constructed, and human-behavioral environments, can be used to incorporate a holistic view of these diverse issues. Explains and illustrates this approach to family concerns often encountered by pastoral caregivers.

Cellular diversity in mouse neocortex revealed by multispectral analysis of amino acid immunoreactivity.

Cortical cells were classified using an unsupervised cluster analysis based upon their quantitative and combinatorial immunoreactivity for glutamate, gamma-aminobutyric acid (GABA), aspartate, glutamine and taurine. Overall, cell class-specific amino acid signatures were found for 12 cellular types; seven GABA-immunoreactive (GABA-IR) populations (GABA1--7), three classes containing high glutamate levels (GLUT1--3) and two putative glial (GLIA1, 2) cell types. From their large somata, associated vertical processes and high glutamate content, the GLUT classes most probably correspond to pyramidal neurons. Two of the GLUT classes demonstrated complementary distributions in different cortical layers, suggesting spatial separation of cells differing in amino acid immunoreactivity. Of the seven GABA classes, two comprised cells with large somata and displayed medium to low glutamate levels. On the basis of size, these two populations may correspond to large basket cell interneurons. Glial populations could be divided into two classes: GLIA1 cells were more frequently associated with blood vessels and GLIA2 cells were more commonly seen in the lower cortical layers. This work demonstrates that signature recognition based upon amino acid content can be used to separate cortical cells into different categories and reveal further subclasses within these categories. This approach is complementary to other methods using physiological and molecular tools and ultimately will enhance our understanding of neuronal heterogeneity.

Contexts for understanding forgiveness and repentance as discovery: a pastoral care perspective.

Conceptualizes forgiveness and repentance from a discovery process paradigm and examines various contexts for understanding and facilitating the forgiveness/repentance process. Suggests a "body/mind/spirit" connection with implications for pastoral care and counseling.

Immune regulation in Cushing's syndrome: relationship to hypothalamic-pituitary-adrenal axis hormones.

Hormones of the hypothalamic-pituitary-adrenal (HPA) axis are connected closely with immune measures. To investigate whether Cushing's syndrome (CS) is associated with immune dysregulation, we compared the percentage of specific lymphocyte subsets as well as natural cell activity (NKCA) in 48 patients with Cushing's syndrome and 48 age- and sex-matched normal controls. Lymphocyte subset analysis included the percentage of lymphocytes expressing CD3 (total T), CD4 (T helper/inducer), CD8 (T suppressor/cytotoxic) and CD56 (NK cell) antigens. Baseline plasma concentrations of cortisol, ACTH and beta-endorphin as well as 24 h urinary-free cortisol (UFC) levels also were determined. Results indicated a decrease in the percentage of CD4+ cells (p < 0.05), an increase in percentage of CD8+ cells (p < 0.05), a decrease in CD4/CD8 ratios (p < 0.01), and a reduction in NKCA (p < 0.05) in patients with CS compared to matched controls. We also found significant negative correlations between NKCA on the one hand and 24 h UFC (p < 0.05) and plasma beta-endorphin (p < 0.05) on the other. These results indicate there is immune dysregulation in CS patients which can be explained in part by an increase in HPA-axis hormones.

Rickets in alpacas (Lama pacos) in New Zealand.

Abstract Rickets was diagnosed in two weaner alpacas showing ill thrift and lameness during the winter of 1992, from a flock at AgResearch Flock House. Affected alpacas had abnormally shaped ribs with occasional healing fractures, irregular thickening of growth plates and metaphyseal haemorrhages. The mean serum phosphorus concentrations of the alpacas fell during June and July, even though lambs grazing the same pasture had normal serum phosphorus concentrations and the phosphorus concentration of the pasture was considered adequate. Vitamin D deficiency may also have contributed to the osteodystrophy. The alpacas had a thick fleece during the winter, and diurnal Vitamin D3 synthesis resulting from solar irradiation is likely to have been minimal, especially considering the reduced sunshine hours recorded during the 1992 winter. Surviving alpacas recovered after treatment with monosodium phosphate and an oral Vitamin D supplement.

Effects of neonatal sympathectomy and capsaicin treatment on bone remodeling in rats.

Bone metabolism may be influenced by the innervation of skeletal tissues. Neuropeptides such as vasoactive intestinal peptide, from sympathetic nerves, and calcitonin gene-related peptide, from sensory nerves, have been implicated as local modulators of bone metabolism. The effect of neonatal sympathectomy and of capsaicin-induced sensory denervation in rats was studied on the following: (i) the radial bone growth and apposition rate in tibiae (normal growth and modeling) and (ii) the percentage of periosteal surface of the mandible occupied by osteoclasts during induced remodeling. Neonate rats were treated with guanethidine, capsaicin, or appropriate vehicle. At seven weeks, maxillary molars were removed to induce remodeling on the buccal surface of the mandible. Animals were killed four days after surgery. Cross-sectional cortical area, medullary area, and periosteal apposition rate were measured by histomorphometry in ground sections of tibiae. The percentage of periosteal surface at the remodeling site occupied by osteoclasts (stained for acid phosphatase) was measured in frozen, undecalcified sections. There was no significant difference in cortical or medullary area or periosteal apposition rate in tibiae between each drug treatment and its control. However, the mandibular bone surface occupied by osteoclasts was increased 45.5% (P less than or equal to 0.005) in animals treated neonatally with guanethidine compared to controls. In contrast, the mandibular surface occupied by osteoclasts was decreased 21.2% (P less than or equal to 0.04) in animals treated neonatally with capsaicin compared to controls. The alteration of bone remodeling (osteoclast surface) by both treatments indicates that sensory and sympathetic nerves play a role in focal metabolism of bone.

Prevalence of bottled water usage by pediatric dental patients: implications for dental health.

Drinking bottled water has become a popular substitute for tap water because of the concern over the contamination of municipal water supplies. The purpose of this study was to determine the percentage of pediatric dental patients drinking bottled water as their primary source of water. The fluoride content of these products was obtained from the distributors and the products were independently analyzed to ensure accuracy. Approximately 10 percent of 1,126 randomly selected patients from a private pediatric dental practice were routinely using bottled water from nine different sources. The fluoride content of these products varied from 0.04 ppm to 1.4 ppm. Independent analysis of the fluoride content of the different brands of bottled water using a microanalyzer with a fluoride-specific electrode indicated that the measured fluoride content was within +/- 0.1 ppm of the distributors' reported fluoride levels. The results of this study found that 16.9 percent of the pediatric patients were receiving less than the optimal level of fluoride and 72.4 percent were receiving greater than the recommended level of fluoride. Ten percent of the patients were being supplemented with additional fluoride tablets by their pediatrician, although the fluoride levels in the bottled water ranged from 0.9 to 1.4 ppm. In order for children to receive the optimal caries-preventive benefit from fluoride, the pediatric dentist needs to question the source of the patients' drinking water routinely. When bottled water is being used, the fluoride content should be obtained from the distributor or submitted for laboratory evaluation for fluoride content.

Orofacial infection of athymic mice with defined mixtures of acyclovir-susceptible and acyclovir-resistant herpes simplex virus type 1.

Infection of athymic mice with defined populations of acyclovir-susceptible (thymidine kinase [TK]-positive) and acyclovir-resistant (TK-deficient or TK-altered) herpes simplex virus type 1 strains was used to simulate herpetic skin disease of the immunocompromised host. In vitro characterization of the defined virus mixtures revealed that the dye uptake method was quite sensitive in the detection of small amounts (3 to 9%) of acylovir-resistant virus. Mice infected with homogeneous virus populations exhibited a good correlation between clinical response and the in vitro drug susceptibility of the infecting virus. Animals infected with defined mixtures of viruses exhibited varied patterns of infection and responses to acyclovir treatment. However, disease severity was useful in predicting the TK phenotype of virus recovered from lesions. Pathogenic, TK-altered virus was responsible for progressive disease in animals receiving low-dose (0.25-mg/ml) prophylactic acyclovir or high-dose (1.25-mg/ml) delayed therapy. Although this mutant was recovered infrequently, it was responsible for clinically significant disease in the animals from which it was isolated.

Long-term acyclovir suppression of frequently recurring genital herpes simplex virus infection. A multicenter double-blind trial.

Normal adults with six or more episodes of genital herpes in the previous year were enrolled in a one-year, multicenter, double-blind trial comparing placebo with 400 mg of acyclovir administered orally twice daily. Patients with episodes during the study were offered 200 mg of acyclovir administered orally five times daily for five days; this allowed comparison of suppressive and episodic treatment. After one year, 227 (44%) of 519 patients receiving suppressive treatment and seven (2%) of 431 receiving placebo (episodic) treatment remained free of recurrences, and the mean numbers of recurrences per year were 1.8 and 11.4, respectively. Among 67 patients who had received suppressive therapy for one year, the mean duration of lesions in the first episode following the discontinuation of treatment was 9.3 days compared with 7.3 days among 45 patients who had received episodic therapy for one year. Treatment was well tolerated, and no changes were noted in the in vitro susceptibility to acyclovir of herpes simplex virus cultured during or after the one-year trial. Continuous or episodic oral acyclovir therapy for one year remained safe and effective.

Efficacy of linoleic acid administered rectally as monoglyceride.

The potential of the rectal route for administration of essential fatty acids (EFA) as monoglyceride (MG) was investigated. EFA-deficient rats were supplemented with 14 mg linoleic acid/day for 3 days. Supplementation was either by oral administration as corn oil, orally as corn oil-derived MG or rectally as MG. The patterns of polyunsaturated fatty acids (PUFA) in liver and serum lipids, characteristic of EFA deficiency, were altered in the direction of normalcy in similar magnitude by all modes of supplementation, indicating that the rectal route may be useful for administration of EFA. The amounts of phospholipids (PL) and free fatty acids (FFA) in liver changed by all modes of administration. The magnitude of change of total PL and of FFA in liver depended upon the chemical form in which linoleic acid was administered and the route of administration, indicating that these factors affect lipid metabolism.

Dietary fats containing concentrates of cis or trans octadecenoates and the patterns of polyunsaturated fatty acids of liver phosphatidylcholine and phosphatidylethanolamine.

The effects of the mixed cis- 18:1 isomers and mixed trans- 18:1 isomers present in partially hydrogenated soybean oil (PHSO) upon the patterns of polyunsaturated fatty acids (PUFA) in liver phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were studied in rats fed concentrates of cis- 18:1 or trans- 18:1 isomers isolated as triacylglycerides from PHSO. The cis- 18:1 and trans- 18:1 concentrates were fed at levels equal to those present in PHSO fed at 17.9% of the diet. All diets contained the required amounts of both linoleic and linolenic acids. The trans- 18:1 concentrate was found to suppress the levels of 20:4 omega 6 and 20:3 omega 9, and to increase the levels of 18:2 omega 6 and 20:5 omega 3 in PC and PE. The cis- 18:1 concentrate suppressed 20:4 omega 6 in PC, 20:5 omega 3 in PC and PE, and 18:2 omega 6 in PC, but increased the levels of 20:4 omega 6 in PE, and 20:3 omega 9 in PC and PE. The cis- 18:1 concentrate was more effective than the trans concentrate in suppressing 22:6 omega 3. The trans- 18:1 concentrate was more effective in suppressing 20:4 omega 6. The trans- 18:1 isomers appear to modify PUFA metabolism by inhibition of PUFA synthesis, whereas the cis- 18:1 isomers appear to complete with 2-position fatty acyl transfer and to inhibit omega 3 PUFA acylation.

Suppression of arachidonic acid in lipids of rat tissues by dietary mixed isomeric cis and trans octadecenoates.

The influence of dietary isomeric cis and trans octadecenoic fatty acids (ICTO) on the metabolism of polyunsaturated fatty acids in rat tissues was studied by feeding a defined diet rich in partially hydrogenated soybean oil. Experimental and control animals received equal and more than adequate amounts of linoleic and linolenic acids. The total phospholipid (PL) fatty acids of liver, heart, testis, brain and sciatic nerve, and the fatty acids of liver triacylglyceride, cholesteryl ester and individual PL classes were analyzed by capillary gas chromatography. The content of arachidonic acid in the total fatty acids of liver lipids of ICTO-fed rats was found to be lower than those from control rats by the following amounts (in percent): total PL, 35; phosphatidylcholine, 50; phosphatidylinositol, 35; phosphatidylserine 26; phosphatidylethanolamine, 12; cholesteryl ester, 55; and tryglyceride 75. PL 18:2 omega 6 and 20:3 omega 6 levels were elevated, suggesting inhibition of the desaturase-elongase enzymes involved in synthesis of arachidonic acid. Synthesis of 20:5 omega 3 and 20:3 omega 9 was accentuated by the ICTO diet, suggesting an omega 6 series-specific inhibition. A nearly perfect negative correlation between cis 12-18:1 and arachidonic acid was found in liver PL.

Parathion alters incubation behavior of laughing gulls.

One member of each pair of incubating laughing gulls at 9 nests was trapped, orally dosed with either 6 mg/kg parathion in corn oil or corn oil alone, and marked about the neck with red dye. Each nest was marked with a numbered stake and the treatment was recorded. A pilot study with captive laughing gulls had determined the proper dosage of parathion that would significantly inhibit their brain AChE activity (about 50% of normal) without overt signs of poisoning. After dosing, birds were released and the nests were observed for 2 1/2 days from a blind on the nesting island. The activities of the birds at each marked nest were recorded at 10-minute intervals. Results indicated that on the day of treatment there was no difference (P greater than 0.05, Chi-square test) in the proportion of time spent on the nest between treated and control birds. However, birds dosed with 6 mg/kg parathion spent significantly less time incubating on days 2 and 3 than did birds receiving only corn oil. By noon on the third day, sharing of nest duties between pair members in the treated group had approached normal, indicating recovery from parathion intoxication. These findings suggest that sublethal exposure of nesting birds to an organophosphate (OP) insecticide, such as parathion, may result in decreased nest attentiveness, thereby making the clutch more susceptible to predation or egg failure. Behavioral changes caused by sublethal OP exposure could be especially detrimental in avian species where only one pair member incubates or where both members are exposed in species sharing nest duties.

Perturbation of the metabolism of essential fatty acids by dietary partially hydrogenated vegetable oil.

Rats were fed purified diets containing (i) partially hydrogenated soybean oil as source of isomeric octadecenoic acids, (ii) hydrogenated coconut oil as source of saturated fatty acids, and (iii) a low level of corn oil as low-fat control. All diets contained 18% of the linoleate requirement. Rat liver and heart phospholipids were analyzed by gas/liquid chromatography for fatty acids, and liver, microsomes were assayed for desaturase (acyl-CoA, hydrogen-donor: oxidoreductase, EC 1.14.99.5) activities. Products of desaturation reactions measured analytically provided more information with greater statistical significance than did the enzymatic assays. Rats fed isomeric octadecenoic acids showed more severe essential fatty acid deficiency than did saturated-fat and control groups. The suppression of linoleate metabolites was largely due to decreased delta 5 and delta 6 desaturase activities. At several levels of linoleate, the deficiency was more severe at the higher level of isomeric octadecenoic acids. Increasing the intake of linoleate to 7.5% of calories did not suppress deposition of isomeric unsaturated acids in tissue lipids.