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Limited non-clinical immunotoxicity data are available in the dog, although this is a major non-rodent species in regulatory safety studies. The present study aimed to test whether widely accepted immunotoxicity endpoints including lymphocyte subset immunophenotyping, the anti-KLH TDAR assay, and histological examination of the main lymphoid organs were reliable to detect immunosuppression induced by cyclosporine and cyclophosphamide in dogs and could, therefore, be used for non-clinical immunotoxicity evaluation in this species. Male and female Beagle dogs were treated orally from Day 1 for 4 weeks with 25 mg/kg cyclosporine daily, or with 2 mg/kg cyclophosphamide on 4 consecutive days each week, or the same volume of drinking water daily. Blood samples were withdrawn pre-test and on Days 11, 18, and 23 to measure standard hematology parameters and analyze lymphocyte subsets. All animals received an intramuscular injection of 5 mg KLH on Day 11. Sandwich ELISA assays were used to quantify anti-KLH IgM and anti-KLH IgG levels in blood samples taken pre-test, on Days 18 and 23, and pre-test, on Days 23 and 28, respectively. At the end of the treatment period, all animals were submitted to histological examination of lymphoid organs, liver, and kidneys. No signs of marked toxicity were observed. No changes in lymphocyte subsets, but markedly decreased primary anti-KLH IgM and IgG responses, and a slightly-to-markedly increased cortex/medulla ratio in the thymus were observed in cyclosporine-treated dogs. Lower total WBC counts correlating with lower total and B-lymphocyte subset and decreased germinal center development in mesenteric lymph nodes, but no changes in primary anti-KLH IgM and IgG responses were observed in cyclophosphamide-treated dogs. These results demonstrate that widely accepted immunotoxicity endpoints can adequately detect the effects of known immunosuppressive drugs in the dog and support the conclusion that it is a relevant animal species for immunotoxicity evaluation.

Purpose: To test a simple electroretinographic protocol on a representative sample of minipigs., Animal Studied: Minipig., Procedures: Electroretinogram recordings were conducted on 162 healthy minipigs (81 males and 81 females) aged 4-6 months. After a 1.5-h light-adaptation period, the animals were anesthetized with general anesthesia. First, binocular full-field photopic electroretinogram recordings were conducted under photopic conditions. Subsequently, scotopic electroretinogram recordings were conducted during dark-adaptation periods every 4 min for a 20-min period. At the end of this period, the maximal combined rod-cone response was recorded by measuring the retinal response to a single high-intensity flash. We used sclerocorneal clip electrodes as active electrodes and needle electrodes as reference and ground electrodes., Results: The a-wave and b-wave peak times and amplitudes have been measured and statistically analyzed. For each of the statistical comparisons, normality and homogeneity of variances were evaluated. No significant gender differences were observed, with the exception of a higher b-wave amplitude for the photopic ERG recordings observed in females when compared to males (48.14 ± 12.909 μV vs. 42.88 ± 10.666 μV; P = 0.005). The process of dark adaptation was evaluated, and the maximal combined rod-cone response was measured (a- and b-waves amplitude and peak time)., Conclusions: We conducted photopic and scotopic electroretinogram recordings from a protocol based on light adaptation followed by dark adaptation using sclerocorneal clip electrodes, which allows quick assembly and examination., (© 2012 American College of Veterinary Ophthalmologists.)

Although a T-dependent antibody response (TDAR) assay is generally recommended as the first-line immune function assay in nonclinical immunotoxicity evaluation, second-line assays such as delayed-type hypersensitivity (DTH) to measure cell-mediated responses can provide helpful additional information. In this study, male Cynomolgus monkeys were injected intramuscularly either once or twice with 1 mg Keyhole Limpet Hemocyanin (KLH) or twice with a commercially available tetanus vaccine (40 IU tetanus toxoid + 0.06 mg aluminum hydroxide). All animals were subsequently challenged by intradermal injections of the same antigen or aluminum hydroxide after 4, 6 and 8 weeks. Clinical reactions at the injection sites were scored 24, 48 and 72 h post challenge. Skin biopsies were taken on completion of the observation period after each challenge for standard histological examination and immunolabeling using CD3 (T lymphocytes), CD19 (B lymphocytes) and CD68 (macrophages) antibodies. Tetanus toxoid induced stronger clinical reactions than KLH, whereas aluminum hydroxide induced no clinical reaction. Perivascular mononuclear cell infiltrates, a histopathological finding consistent with a DTH reaction, were seen after all challenges with tetanus toxoid or KLH, but not with aluminum hydroxide. Immunohistochemistry evidenced the presence of T lymphocytes and macrophages within these infiltrates. These results suggest that tetanus toxoid adjuvanted with aluminum hydroxide can induce a consistent DTH response for use as a model of cell-mediated response in Cynomolgus monkeys.

Parabens (PB) are preservatives used in food, drugs and personal care products preventing microbial and fungal contamination. We investigated ADME profiles of [14C]-methyl-, propyl- or butylparaben (MP, PP, BP) following single oral, dermal or subcutaneous (BP) doses at 100 mg/kg to Sprague-Dawley rats. Plasma Cmax and AUC values after oral or subcutaneous doses were 4- to 10-fold higher relative to respective values after dermal administration. tmax ranged from 0.5, 2 or 8 h after oral, subcutaneous or dermal administration, respectively. MP produced higher blood Cmax and AUC levels relative to those after PP or BP. Following oral or subcutaneous administration, urinary excretion was predominant (>70%, mainly during the first 24 h), less than 4% were eliminated in the feces, 2% were retained in the tissues and carcasses. Following dermal application, >50% of the dose was unabsorbed, 14-27% or <2% were respectively excreted in the urine or feces, respectively. Overall, parabens were well absorbed after oral and subcutaneous, and partially absorbed after dermal administration. All administration routes produced a single peak in the plasma, corresponding to that of para-hydroxybenzoic acid (PHBA) suggesting that PB produce no significant systemic exposure of mammalian organisms after oral, topical or subcutaneous administration., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The ovariectomized old cynomolgus monkey is a recognized model of human osteoporosis, and the same species can be used for the assessment of the efficacy and potential toxicity of agents intended to prevent or treat osteoporosis. Several assays have been developed that can measure the same biochemical markers of bone turnover as are used in human patients for the diagnosis and treatment follow-up of bone-related diseases, including osteoporosis. The aim of the present study was to describe the results obtained with these assays in normal control monkeys, their variations with age and sex, and their sensitivity in monitoring the bone turnover induced by ovariectomy in old skeletally mature cynomolgus monkeys. Seven old cynomolgus monkeys were bilaterally ovariectomized and 13 age-matched monkeys were sham-operated. Bone mineral density and biochemical markers were measured before and at regular intervals after surgery for up to 20 months. Total alkaline phosphatase (total ALP), bone-specific alkaline phosphatase isoenzyme (bone ALP) and osteocalcin (OC) were highly correlated to the decrease in bone mineral density (BMD) induced by ovariectomy. Deoxypyridinoline (DPD) measured by enzyme-linked immunoassay was insensitive to the bone resorption induced by ovariectomy, but cross-linked N-telopeptide (NTX-I) was higher in ovariectomized monkeys than in control monkeys. These results demonstrate that reliable biochemical parameters are available to adequately monitor and provide insight into osteoclastic bone resorption and osteoblastic bone formation, the two components of bone turnover in this animal model, and can thus be used to assess the efficacy and toxicity of potential therapeutic agents.

The investigations on the activity of respiratory ciliae in vivo are of considerable interest for the study of physiopathological disorders resulting from chronic infection or inhaled pollutants. We present an image analysis method for the measurement of ciliary beat frequency on mammal or human in vivo samples. Videofilm sequences of samples placed under the microscope were analyzed frame by frame by image analysis. The operator defines on the screen several areas of interest on the cell apex. The variation of grey levels in each areas allows the calculation of ciliary beat parameters. This method permits one to investigate the effects of pharmacological agents or noxious pollutants on ciliae beating and the physiological causes underlying them. Our results on rabbits were in agreement with the data published in literature, showing no significant difference between intratracheal frequencies obtained at medium and lower levels. The upper level near the cracoid cartilage has shown a greater variability than the other two levels with higher beating frequencies, a difference of likely physiological importance.

Quantitative image analysis is used to measure the inotropic and chronotropic effects of drugs on cultured heart cells maintained at 37 degrees C on the stage of an inverted light microscope, and sequentially superfused with control and treatment media. The beating of the cardiac myocytes is evaluated by simultaneously selecting up to eight areas, including cell edges, from digitized video image. The sizes and positions of these areas are controlled by the operator. To analyze the motion of cell edges in each area, the computer measures the shift of the mass center of pixels' grey levels. Finally, a few parameters are calculated for the eight areas and displayed graphically. In order to assess treatment effects, appropriate statistical tests are performed on the data. Image analysis is an efficient screening test for evaluating the pharmacologic or toxic effects of a substance on isolated or cultured cardiac myocytes from various species.

The action of two epimers of a new vinblastine derivative that differ in their in vivo antitumor activity and their cytotoxicity was studied in vitro in brain microtubule proteins. These two compounds, called S-12363 and S-12362, could not be distinguished from one another or from other active vinca alkaloids by their ability to prevent microtubule assembly. However, they differed strongly both from one another and from vincristine and vinblastine in their ability to induce the formation of tubulin paracrystals and in the stability of the paracrystals following temperature shifts from 0 degree to 37 degrees C and vice versa. The most potent drug, S-12363, induced considerable tubulin aggregation, which was even more pronounced than that observed in the presence of vincristine. Previous results have shown that S-12363, in contrast to vincristine, induces no neurotoxic effects. This observation is in disagreement with a direct relationship between tubulin aggregation and neurotoxicity.

The results of 265 trochanteric fractures in the elderly treated between 1980 and 1983 have been analysed. Among them, 136 were treated by Ender's nailing, 110 by prostheses and 19 by internal fixation. In an earlier study, Ender's nailing had been used in 75 p. 100 of cases. The conclusions were that the increasing use of a prosthesis has brought about an improvement in the results-80 p. 100 satisfactory in the present series compared with 54 p. 100 in the preceding one. The mortality rate was comparable after Ender's nailing and prostheses. Ender's nailing is indicated in stable fractures and, in unstable fractures, internal fixation is indicated in very active persons, Ender's nailing in non-ambulatory patients and prostheses in elderly patients with sufficient walking ability.

The neuroendocrine factors synthetized by various regions of the nervous system and which have inhibiting or stimulating effects can control the male and female physiology of malacostraca via sexual hormones and moulting hormone. There are between decapods and peracarids functional homologies but some differences can be noticed as to the localization of these antagonistic neurosecretory systems. The functioning of these systems is regulated by external factors such as the photoperiod and the temperature. The interactions between these different neurohormones, the sexual hormones and the moulting hormone are discussed.

The authors studied 65 men who had a varicocele by semen analysis, estimating the levels of FSH, LH, testosterone and prolactin and in some cases the rise in FSH using GnRH. 59 cases were treated by operation and in each of these testicular biopsy was carried out on both sides. Indications are given as to the duration in years of the infertility, the localisation of the varicoceles and the preoperative semen analysis. It would appear that the FSH reading is most important. When it is more than 3.5 ng/ml no pregnancy occurred but when the level was within the mean range of 2.8 ng/ml the chances of pregnancy were higher. It came out at 43% of spouses becoming pregnant within 32 months. The authors make a plea for infertility in both partners to be treated by the same specialist.

Butel's hip plate was used for osteosynthesis of 241 fractures of upper end of femur (100 true cervical - 141 trochanteric and subtrochanteric fractures). Results for true cervical fractures were assessed as satisfactory in 89.5% of cases, with only 3 pseudarthroses and 4 cases of femoral head necrosis (2 septic, 2 aseptic), a total complication rate of 7.5%. These clinical results confirm the value of screw fixation at several cephalic anchorage sites (demonstrated biomechanically) in true cervical fractures. Results in trochanteric and subtrochanteric fractures were rated as satisfactory in 86.9% of cases, complications including 2 ruptures of plate, 1 sepsis and 4 early loosening of plate. The latter sequela was avoided by an improved choice of indication for the procedure and by substitution of this compound material (screw-plate) for a monobloc piece ("anti-loosening cervicocephalic screw apparatus"), in compound fractures of trochanter.

A reversal of sexual phenotype resulting in the appearance of neo-males and neo-females occurs in the populations of two marine subspecies of Idotea balthica. Sex inversions are rare in I. b. basteri but more frequent in I. b. tricuspidata. By crossing these two subspecies, the rate of neo-males (ti) was considerably increased, allowing us to study the determination of sex inversion. The I. b. tricuspidata used in this study came from the offspring (27 p. 100 neo-males) of one gravid albafusca female (A1) collected in La Rochelle (LR); these offspring had been studied for five generations. The I. b. basteri came from offspring of females collected in Marseille (M), Racou (eastern Pyrenees) and Tunis. The chi 2 or Mann-Whitney test was used to compare the results of the different crosses. The female Idotea balthica is heterogametic (WZ); the male is homogametic (ZZ). The loci of the genes which determine the colored phenotypes, albafusca (A) and bilineata (B), are situated on the W heterochromosome. The fundamental phenotype, uniformis (U), is the wild type, according to classical genetics. The males used were either ZZ and uniformis or WZ (neo-males) and albafusca. 1) Ten crosses: female A basteri (M) X male U tricuspidata (LR) gave a variety of offsprings (chi 2 = 56 - d.f. = 9), two having an A1-like ti and the other eight no or very few neo-males. 2) Backcrosses (from CRF2 to CRF5) between hybrid females and U La Rochelle males: in CRF2, the ti increased significantly and in CRF3, CRF4 and CRF5 it reached the ti of the A1 lineage. 3) In the female F1 X male UM backcrosses, the ti was nearly zero, thus being comparable with the basteri lineage of Marseille. 4) When female ALR X male UM of some lineages was crossed, the expressivity of the phenotype A was reduced (from A to U) only in neo-males. The penetrance of gene A did not vary significantly in seven crosses from F1 to CRF2. 5) The crossing of male male ULR with female female basteri B or U from various populations showed that only some B and U females were likely to favor sex inversion in their daughters; the resulting ti was ten times higher with A females. These results led to the following conclusions: --crosses 1, 2 and 3 showed that reinforcement of the tricuspidata genotype of La Rochelle, by means of a regular addition of mainly autosomal genes, induced an increase in the ti, whereas reinforcement of the basteri genotype induced a decrease in that rate. The results supported the hypothesis of a polyfactorial determination of sex involving autosomal, or partly sex-linked, factors acting as inhibitors of the major female gene(s); --crosses 4 showed that the reduction of gene A expressivity was always associated with the sex inversion of genetic hybrid females; that process also seemed to be controlled by some autosomal or partly sex-linked recessive genes, different from those of the preceding complex and acting as inhibitors of the major female-determining factor(s) situated on the W heterochromosome...

One hundred and seventeen shoulders in 113 patients were treated surgically for recurrent anterior dislocation of the shoulder by the Latarjet procedure - transplantation of the coracoid process to the anterior border of the glenoid fossa. 67 p. 100 of the patients engaged in sport and 33 p. 100 did not. The usual lesions found at operation were separation of the capsule from the anterior rim of the glenoid in 59 p. 100, a lesion of the glenoid labrum in 65 p. 100 and damage to the glenoid fossa short of a fracture in 18 p. 100. Postoperative complications were rare and usually mild, except in two cases of osteoporosis of the coracoid transplant and two fractures of the bone block. There was slight limitation of lateral rotation in 48 p. 100, of abduction in 14 p. 100 and of medial rotation in 4 p. 100. The results at follow-up were rated as excellent in 68 p. 100 (72 cases), good in 21.7 p. 100 (23 cases), with slight pain and limitation of lateral rotation of less than 20 p. 100 and fair in 6.6 p. 100 (7 cases). In these cases, the Latarjet procedure had been performed for postoperative recurrence after a previous surgical procedure. There were 4 poor results.

Two models have been chosen to study the effect of 2-amino-6-trifluoromethoxy benzothiazole (PK 26124) on excitatory amino acid neurotransmission: the pool of cyclic guanosine monophosphate (cGMP) in the cerebellum and the release of acetylcholine in the striatum and olfactory tubercles. The release of acetylcholine induced by N-methyl-DL-aspartate in the striatum and olfactory tubercles was antagonized by PK 26124 which was less potent on the release of acetylcholine induced electrically. The increase in levels of cGMP in the cerebellum induced by excitatory amino acids such as glutamate and quisqualate was antagonized by PK 26124, but the drug was inactive against N-methyl-DL-aspartate, L-aspartate, kainate and cysteine sulphinate. In vivo it antagonized the increases of cGMP in the cerebellum elicited by all these excitatory compounds. All these results are compatible with a possible antagonism by PK 26124 of the excitatory amino acid neurotransmission and may explain its anticonvulsant properties.

A symbiotic bacteroid matroclinously transmitted is present in the ovocytes and the follicular cells of the sub-species Porcellio dilatatus petiti of the Saint-Honorat island, as well as in all hybrid females born from a petiti mother. It is absent in the Porfellio dilatatus dilatatus females (population of Poitiers) and this seems to cause the lithality in the periblastula stage -- as a result of abnormal anaphases--of practically all the embryos of the crossing : male petiti x female dilatatus.

The males with female genital apertures of the Niort population of A. vulgare are genetic males that have been made intersexual by two factors which are probably epigenetic : one is feminizing, the other masculinizing. The transmission of the masculinizing factor seems to be controlled by a genetic mechanism which, in the crossing of an intersexe with a genetic female, makes half the female issue turn into neo-males and intersexes.

2-Amino-6-trifluoromethoxy benzothiazole (PK 26124) prevented convulsions induced in rodents by maximal electroshock, inhibitors of the synthesis of gamma-aminobutyric acid (GABA) and ouabain, but was inactive against seizures provoked by GABA antagonists, unlike diazepam, chlordiazepoxide, phenobarbital and valproic acid. 2-Amino-6-trifluoromethoxy benzothiazole prevented seizures induced by sound stimuli in DBA/2 mice (ED50 = 0.66; 2.1 and 4.1 mg/kg, i.p. according to the seizure component), postural seizures in El mice (ED50 = 7.5 mg, i.p.) and seizures induced by photic stimulation in the baboon, Papio papio, at 4 and 8 mg/kg (i.v.). This spectrum of anticonvulsant activity closely resembles that reported previously for dicarboxylic amino acid antagonists. Indeed, PK 26124 prevented seizures induced by L-glutamate (ED50 = 8.5 mg/kg, i.p.) or by kainate (ED50 = 9.25 mg/kg, i.p.) and tremors induced by harmaline (ED50 = 2.5 mg/kg, i.p.) In these tests diazepam was inactive (L-glutamate) or as potent as PK 26124 (kainate, harmaline), whereas it was 10-20 times more potent than PK 26124 against seizures induced by inhibitors of the synthesis of GABA. Together, these data suggest that PK 26124 possesses antagonistic properties of excitatory dicarboxylic amino acids, which may contribute to its anticonvulsant action.

In thelygenous lines of Armadillidium vulgare, neo-females and intersex males (iM) with feminizing symbiotic bacteria are not masculinized by an extract from iM androgenic gland, which, however, masculinizes bacterialess genetic females. Injection of iM hemolymph extract masculinizes these genetic females. This indicates that androgenic hormone is present in iM hemolymph. Lack of androgenic hormone activity in thelygenous lines is supposed to result from the action of bacteria on the androgenic hormone receptors. Since a temporary recovery of the male differentiation of iM can be induced by implantation of different parts of central nervous system, bacteria effect is probably indirect, through an action on a neurosecretory system, perhaps one of those controlling the functioning of the androgenic gland.

A review of complications of 374 fractures of the proximal femoral end, treated by Ender's nailing was conducted: 255 cases only were available because of 46 early deaths and 73 patients without enough follow up. Complications were: 31 superior nails' perforations, 15 downward migrations, 17 dislocations of osteosynthesis and 19 rare complications. The authors believe that Ender's nailing is a great improvement, when compared with classical methods of osteosynthesis or prosthetic replacements. However, a perfect technical execution is necessary to avoid mechanical complications.