Treatment-Emergent Influenza Virus Polymerase Acidic Substitutions Independent of Those at I38 Associated With Reduced Baloxavir Susceptibility and Virus Rebound in Trials of Baloxavir Marboxil.

Influenza viruses harboring treatment-emergent I38F/M/N/T substitutions in the polymerase acidic (PA) endonuclease exhibited reduced susceptibility to baloxavir and were associated with virus rebound and variable clinical response in clinical trials. US regulatory review of registrational trial data also identified treatment-emergent PA substitutions E23K in A/H1N1 viruses and E23G/K, A37T, and E199G in A/H3N2 viruses, which conferred reduced susceptibility to baloxavir, although to a lesser degree than I38F/M/N/T substitutions, and were associated with virus rebound. Although these non-I38 substitutions emerged less frequently than substitutions at I38, they represent alternate pathways to baloxavir virologic resistance and should be monitored accordingly.

The role of learning by a predator, Rivulus hartii, in the rare-morph survival advantage in guppies.

Negative frequency-dependent selection (NFDS), where rare types are favoured by selection, can maintain diversity. However, the ecological processes that mediate NFDS are often not known. Male guppies (Poecilia reticulata) exhibit extreme diversity of colour patterning and, in a previous field experiment, rare morphs had a survival advantage. Here, we test the hypothesis that predators impose NFDS because they are efficient at capturing familiar prey morphs, but are less efficient at capturing unfamiliar morphs. Over a series of trials, we presented Rivulus hartii, a natural predator of guppies, with male guppies with the same colour patterning (A trials); then, for a second series of trials, we presented the rivulus with guppies with a new colour pattern (B trials). The success of rivulus at capturing guppies on the first attack increased over successive A trials. First attack success decreased significantly for the early B trials, and then increased during successive B trials, eventually reaching the same level as in the best A trials. This experiment demonstrates that learning, perhaps through long-term search image formation, plays a role in predation success on familiar vs. unfamiliar prey morphs. These results support the hypothesis that predator learning contributes to the maintenance of the extreme male guppy polymorphism seen in nature.

Corium tissue expression of genes associated with inflammation, oxidative stress, and keratin formation in relation to lameness in dairy cows.

Objectives were to (1) determine the feasibility of performing hoof biopsies without impairing locomotion; (2) evaluate the feasibility of using biopsied tissue for quantitative PCR; and (3) compare relative gene expression among claws for several target genes. Biopsies were performed on 6 Holstein cows, yielding 4 tissue specimens per cow from front leg, right limb, and medial claw (claw position 3); rear leg, left limb, and lateral claw (claw position 5); and rear leg, right limb, medial claw (claw position 7). Cows were monitored for lameness daily for 7 d post-biopsy and then weekly for 8 wk. Histopathological analysis confirmed that tissue collected was from between the stratum corneum and dermis. Biopsied tissue was used for RNA extraction, including evaluation of yield and purity. The profile by claw position of 19 genes with key functions in cell differentiation, proliferation, inflammation, and keratin formation was assessed via quantitative reverse transcription-PCR. Other than transient disturbances in locomotion score in some cows during 2 to 4 d post-biopsy, no signs of pain, locomotion impairment, or clinical lameness were observed post-biopsy. Total RNA yields averaged 259.7±100, 447.8±288, and 496.4±118 µg/mg of tissue for claw positions 3, 5, and 7, respectively. The biopsy procedure was successful for obtaining corium for gene expression. Among 5 keratin proteins analyzed, only keratin 5 was expressed. Transcripts related to inflammation and oxidative stress (STAT3, MYD88, SOD2, and TLR4) were among the more abundant in corium tissue, but expression did not differ between claws. Biotinidase (BTD) expression was greater in claw 3 versus claw 5, whereas the ligand-activated nuclear receptor retinoic acid receptor-α (RXRA) was greater in claws 3 + 5 compared with claw 7. Overall, results from this pilot study revealed modest differences at the transcriptome level, suggesting that biotin availability and lipid metabolism differ between claw positions, whereas inflammation and oxidative stress seem to play an important role across claws. More comprehensive studies of the hoof transcriptome are required to improve our understanding of the mechanisms that link environmental and dietary factors to development of lameness.

MOP and NOP receptor interaction: Studies with a dual expression system and bivalent peptide ligands.

Opioids targeting mu;µ (MOP) receptors produce analgesia in the peri-operative period and palliative care. They also produce side effects including respiratory depression, tolerance/dependence and addiction. The N/OFQ opioid receptor (NOP) also produces analgesia but is devoid of the major MOP side effects. Evidence exists for MOP-NOP interaction and mixed MOP-NOP ligands produce analgesia with reduced side effects. We have generated a HEKMOP/NOP human expression system and used bivalent MOP-NOP and fluorescent ligands to (i) probe for receptor interaction and (ii) consequences of that interaction. We used HEKMOP/NOP cells and two bivalent ligands; Dermorphin-N/OFQ (MOP agonist-NOP agonist; DeNO) and Dermorphin-UFP101 (MOP agonist-NOP antagonist; De101). We have determined receptor binding profiles, GTPγ[35S] binding, cAMP formation and ERK1/2 activation. We have also probed MOP and NOP receptor interactions in HEK cells and hippocampal neurones using the novel MOP fluorescent ligand, DermorphinATTO488 and the NOP fluorescent ligand N/OFQATTO594. In HEKMOP/NOP MOP ligands displaced NOP binding and NOP ligands displaced MOP binding. Using fluorescent probes in HEKMOP/NOP cells we demonstrated MOP-NOP probe overlap and a FRET signal indicating co-localisation. MOP-NOP were also co-localised in hippocampal tissue. In GTPγ[35S] and cAMP assays NOP stimulation shifted the response to MOP rightwards. At ERK1/2 the response to bivalent ligands generally peaked later. We provide evidence for MOP-NOP interaction in recombinant and native tissue. NOP activation reduces responsiveness of MOP activation; this was shown with conventional and bivalent ligands.

Extraction of cheek teeth using a lateral buccotomy approach in 114 horses (1999-2009).

REASON FOR PERFORMING THE STUDY: There are few published data regarding the success rates of cheek tooth (CT) removal by lateral buccotomy in the horse. OBJECTIVES: A retrospective study of 114 horses admitted to 2 private equine referral hospitals over a 10 year period (1999-2009), which underwent CT removal via a lateral buccotomy. METHODS: Hospital records were analysed and details including case details, presenting complaint and results of all diagnostic tests and surgical reports were documented. Information obtained during post operative reassessment was also available for analysis. Long-term follow-up information (>2 months) was obtained for 112 horses. RESULTS: Short-term complications (<2 months) occurred in 24/77 horses (31%) undergoing exodontia of the maxillary CT and 10/37 horses (27%) involving mandibular CT, with the majority arising from partial wound dehiscence and infection following 16 extractions (47%). All healed well by second intention. Other complications included both temporary (n = 6) and permanent (n = 3) facial nerve paralysis, myositis (n = 4) and the inadvertent establishment of an oroantral fistula (n = 4). One myositis case was subjected to euthanasia 24 h post operatively. Five horses had persistent sinusitis following surgery due to dental remnants (n = 2) and excessive packing material (n = 1) found in the sinuses. No inciting cause could be found in the remaining 2 horses. One further horse suffered a fatal cardiac arrest at induction of anaesthesia. Of all horses, 92% operated on had returned to their previous level of work after >2 months with no complications. CONCLUSION: Horses with CT removal by a lateral buccotomy have a reasonable prognosis for long-term outcome. POTENTIAL RELEVANCE: Performing a lateral buccotomy is a justified treatment alternative for the surgical removal of equine CT following unsuccessful attempts by standing oral extraction, offering advantages over alternatives such as retropulsion, endodontic therapy and periapical curettage.

Factors associated with change and stability in adherence to muscle-strengthening guidelines among young Australian adults: A longitudinal study.

OBJECTIVES: The 2014 Australian Physical Activity and Sedentary Behaviour Guidelines (Adults) recommend muscle-strengthening activities ≥2 days/week. This study aimed to identify factors associated with 5-year change and stability in adherence to these guidelines. DESIGN: Two adult follow-ups of the Childhood Determinants of Adult Health (CDAH) Study. Participants (n = 1510) were 26-36 years (CDAH-1, 2004-06) and 31-41 years (CDAH-2, 2009-11). METHODS: Information on muscle-strengthening activities, sociodemographics, health, physical activity and sedentary behaviour was collected. Participants reporting muscle-strengthening activities ≥2 days/week 'met guidelines', with change and stability categorised as 'persistent adherence', 'increasing adherence', 'decreasing adherence' and 'persistent non-adherence'. Differences in sociodemographic, health and behavioural factors were analysed using log multinomial regression. RESULTS: Between 15-21% of women (CDAH-1: 14.5%, 95% confidence interval = 12.5-16.9; CDAH-2: 20.7%, 95% confidence interval = 18.3-23.4) and ~21% of men (CDAH-1: 22.2%, 95% confidence interval = 19.0-26.0; CDAH-2: 21.0%, 95% confidence interval = 17.8-24.7) met muscle-strengthening guidelines, but only 8.5% (95% confidence interval = 7.2-10.1) of participants were persistently adherent. Remaining in or moving from a major city, CDAH-1 weight status, cumulative self-rated health and vigorous physical activity were positively associated with persistent adherence (relative risk range = 1.51-3.92), while female gender, becoming partnered and having children at any timepoint were negatively associated with persistent adherence (relative risk range = 0.38-0.58). CONCLUSIONS: Adherence - particularly persistent adherence - to muscle-strengthening guidelines in this sample was low. Gender, marital status, weight status, BMI, self-rated health, urban-rural status, parental status, physical activity and sedentary behaviour were associated with adherence, and should be considered in intervention development to maximise effectiveness.

Nonallelic behavior of rabbit variable-region allotypes.

Group a allotypes not detected by qualitative typing or anticipated from breeding data (latent allotypes) were detected at low levels in 50% of normal rabbit sera tested. The latent allotypes, which were serologically identical to allotypes of pooled IgG, were detected in sera from rabbits with all possible combinations of group a allotypes and their occurrence in individual rabbits was transitory and sporadic. These findings give reason to question the assumption that the synthesis of immunoglobulin allotypes is directed by allelic structural genes.

Selection at the MHC class IIB locus across guppy (Poecilia reticulata) populations.

The highly diverse genes of the major histocompatibility complex (MHC) are important in the adaptive immune system and are expected to be under selection from pathogens. Thus, the MHC genes provide an exceptional opportunity to investigate patterns of selection within and across populations. In this study, we analyzed genetic variation at the MHC class IIB gene and six microsatellite loci across 10 populations of guppies (Poecilia reticulata) in the northern range of Trinidad. We found a high level of diversity at the MHC, with a total of 43 alleles in 142 individuals. At the population level, we found that neutral evolution could not fully account for the variability found at the MHC. Instead, we found that MHC F(ST) statistics were lower than F(ST) derived from the microsatellite loci; 33 of 45 population pairwise estimates for the MHC were significantly lower than those for the microsatellite loci, and MHC F(ST) estimates were consistently lower than those predicted by a coalescent model of neutral evolution. These results suggest a similar selection acting across populations, and we discuss the potential roles of directional and balancing selection. At the sequence level, we found evidence for both positive and purifying selection. Furthermore, positive selection was detected within and adjacent to the putative peptide-binding region (PBR) of the MHC. Surprisingly, we also found a purifying selection at two sites within the putative PBR. Overall, our data provide evidence for selection for functional diversity at the MHC class IIB gene at both the population and nucleotide levels of guppy populations.

Quantitative evaluation of subchondral bone injury of the plantaro-lateral condyles of the third metatarsal bone in Thoroughbred horses identified using nuclear scintigraphy: 48 cases.

REASONS FOR PERFORMING STUDY: Increased radio-isotope uptake (IRU) in the subchondral bone of the plantaro-lateral condyle of the third metatarsus (MTIII) is a commonly reported scintigraphic finding and potential cause of lameness in UK Thoroughbred racehorses in training and has not been fully documented. OBJECTIVES: To characterise lameness attributable to IRU of the subchondral bone of MTIII, compare the scintigraphic findings of these horses with a normal population and evaluate the use of scintigraphy as an indicator of prognosis. HYPOTHESIS: IRU will be in significantly higher in horses with subchondral bone injury and will be related to prognosis and future racing performance. METHODS: Data were analysed from 48 horses in which subchondral bone injury of the plantaro-lateral condyle of MTIII had been diagnosed using nuclear scintigraphy and that met the inclusion criteria. Data recorded included age, sex, trainer, racing discipline, lameness assessment, treatment regimes, radiographic and scintigraphic findings, response to diagnostic analgesia where performed and racing performance pre- and post diagnosis. Region of interest (ROI) counts were obtained for the plantar condyle and the mid diaphysis from the latero-medial view, the ratio calculated and then compared with a control group of clinically unaffected horses. RESULTS: The mean condyle mid-diaphysis ROI ratio was significantly (P<0.001) higher in the affected population and with positively correlation (P = 0.024) with the level of lameness. The presence of radiographic findings had no significant effect on the ROI ratio. CONCLUSION: Subchondral bone injury of the plantar lateral condyles of MTIII is a significant cause of lameness in UK Thoroughbred racehorses. Nuclear scintigraphy is a useful diagnostic imaging modality in the detection of affected horses but is a poor indicator of prognosis for the condition. POTENTIAL RELEVANCE: Better understanding of the clinical manifestations, diagnosis of and prognosis for subchondral bone injury will benefit the Thoroughbred industry in the UK.

An investigation of the relationship between race performance and superficial digital flexor tendonitis in the Thoroughbred racehorse.

REASONS FOR PERFORMING STUDY: There is limited information regarding the number of races and the period for evaluation of outcome which is critical for assessment of SDF tendonitis treatments. OBJECTIVE: To evaluate the re-injury rate and racing performance of Thoroughbred racehorses that sustain SDF tendonitis in relation to matched controls in terms of number of races post treatment and maximum racing performance ratings before and after injury. STUDY DESIGN: Clinical records and racing histories of 401 racehorses with a first occurrence of SDF tendonitis diagnosed by ultrasonography. Controls were of the same age, sex and were horses training in the same establishment at the time of injury as the case horses and where the trainer reported that the horse had not had a previous SDF tendon injury or treatment. RESULTS: Eighty percent of both case and control horses returned to racing after the date of injury, and the re-injury proportion within 3 years of treatment was 53%. The difference in Racing Post Rating((max)) (RPR((max))) and the Racing Post Rating in the race immediately before the treatment date was significantly smaller in case horses (mean = 9.6 lbs; range = 0-75) compared to control horses (mean = 17.0 lbs; range = 0-79). No significant decrease in RPR((max)) was noted post injury. No difference between case and control horses was found for return to racing and racing 3 times, but control horses were significantly more likely to compete 5 races post treatment date than case horses. CONCLUSION AND CLINICAL RELEVANCE: Injury was associated with an individual's pre-injury maximum performance level and return to racing and completion of 3 races are not useful indicator of the outcome of horses with SDF tendonitis. The assessment of the outcome of horses with an SDF injury in a population of racehorses using the number of races post injury requires a minimum of 5 races post injury to be a useful indicator. Further, a re-injury proportion in a population of horses in training for 3 years post treatment.

Linear body measurements and productivity of subtropical Holstein-Friesian dairy calves.

OBJECTIVE: The study aimed to describe growth parameters and to quantify the association between linear body measurements as predictors of liveweight (LW) of Holstein-Friesian (HF), and HF crossbred dairy calves in Queensland. A secondary objective was to quantify the effect of disease events on LW change. STUDY DESIGN: Longitudinal study. METHODS: Fortnightly LW, hip height (HH), hip width (HW) and heart girth (HG) measurements were recorded from 16 male and 28 female HF calves from birth until weaning. The association between linear body measurement and the effect of a disease event on LW change were explored using linear mixed-effects modelling with random intercepts and random slopes. RESULTS: HG was the best body measure used individually as a predictor of calf LW (R2 = 82%; P < 0.001), while the combined use of HG, HW and HH was the most accurate predictor of calf LW between birth and weaning (R2 = 90%; P < 0.001). HW, average feed intake and total feed intake were significantly affected by disease events (P < 0.05). On average, total average LW loss associated with a single pneumonia event was estimated at 14.6 kg (95% CI = 10.5 to 18.7 kg; P < 0.05). CONCLUSIONS: Calves of this study performed at a level consistent with the previously published reports. Growth performance was significantly compromised by pneumonia. HW was found to be the least predictive individual measure, and the combined use of HH, HW and HG had the most accurate prediction of calf liveweight from birth to weaning.

Synthetic CD4 peptide derivatives that inhibit HIV infection and cytopathicity.

Synthetic peptide segments of the CD4 molecule were tested for their ability to inhibit infection of CD4+ cells by the human immunodeficiency virus (HIV) and to inhibit HIV-induced cell fusion. A peptide mixture composed of CD4(76-94), and synthesis side products, blocked HIV-induced cell fusion at a nominal concentration of 125 micromolar. Upon high-performance liquid chromatography, the antisyncytial activity of the peptide mixture was found not in the fraction containing the peptide CD4(76-94) itself, but in a side fraction containing derivatized peptide products generated in the automated synthesis. Derivatized deletion and substitution peptides in the region CD4(76-94) were used to demonstrate sequence specificity, a requirement for benzyl derivatization, and a core seven-residue fragment required for antisyncytial activity. A partially purified S-benzyl-CD4(83-94) peptide mixture inhibited HIV-induced cell fusion at a nominal concentration of less than or equal to 32 micromolar. Derivatized CD4 peptides blocked cell fusion induced by several HIV isolates and by the simian immunodeficiency virus, SIV, and blocked infection in vitro by four HIV-1 isolates with widely variant envelope gene sequences. Purified CD4(83-94) dibenzylated at cysteine 86 and glutamate 87 possessed antisyncytial activity at 125 micromolar. Derivatization may specifically alter the conformation of CD4 holoreceptor peptide fragments, increasing their antiviral efficacy.

MHC class IIB additive and non-additive effects on fitness measures in the guppy Poecilia reticulata.

The genetic architecture of fitness at the class IIB gene of the major histocompatibility complex (MHC) in the guppy Poecilia reticulata was analysed. Diversity at the MHC is thought to be maintained by some form of balancing selection; heterozygote advantage, frequency-dependent selection or spatially and temporally fluctuating selection. Here these hypotheses are evaluated by using an algorithm that partitions the effect of specific MHC allele and genotypes on fitness measures. The effect of MHC genotype on surrogate measures of fitness was tested, including growth rate (at high and low bulk food diets), parasite load following a parasite challenge and survival. The number of copies of the Pore_a132 MHC allele was inversely related to infection by Gyrodactylus flukes and it appeared to be positively related to faster growth. Also, genotypes combining the Pore_a132 or other relatively common alleles paired with rare MHC alleles produced both advantageous and detrimental non-additive effects. Thus, the genetic architecture underlying fitness at the MHC is complex in the P. reticulata.

The use of race winnings, ratings and a performance index to assess the effect of thermocautery of the soft palate for treatment of horses with suspected intermittent dorsal displacement. A case-control study in 110 racing Thoroughbreds.

REASONS FOR PERFORMING THE STUDY: There have been no reports of the efficacy of thermocautery of the soft palate (TSP) assessed objectively as a treatment of intermittent dorsal displacement of the soft palate (DDSP). OBJECTIVE: To compare: racing performance of horses that underwent thermocautery of the soft palate with matched controls; and 'Racing Post ratings' (RPR) with prize money won (RE) and a performance index (PI) for each of the horses in the study. HYPOTHESIS: Thermocautery of the soft palate has no beneficial effect on racing performance and the 3 measures of performance are significantly related. METHODS: The inclusion criteria were fulfilled by 110 horses and each was matched with 2 controls. Changes in performance were compared statistically. RPR, RE and PI were analysed using a regression model. RESULTS: The percentage of horses that improved in performance following the procedure was 28-51% for the 3 measures of performance, compared to 21-53% for the matched controls. There was no significant effect of the procedure on the changes in RPR or RE. There was a significant effect of the procedure on the change in PI (P=0.015) with more treated horses achieving an improved PI and fewer acquiring a worse PI than matched control horses. The measures of performance showed significant correlation. CONCLUSIONS: Thermocautery of the soft palate alone may not be the most efficacious treatment of DDSP. POTENTIAL RELEVANCE: Production of a reliable measure of racehorse performance may be possible.

Parabens generate reactive oxygen species in human spermatozoa.

Parabens are used as antimicrobial preservative agent in many commercial products including cosmetics and pharmaceuticals. Weak oestrogenic and antiandrogenic activities have been attributed to parabens in in vitro and in vivo studies. In this study, human spermatozoa were exposed to different concentrations of an equimolar paraben mixture containing methyl, ethyl, propyl and butylparaben as well as to methylparaben alone at a concentration that is typical of commercially available vaginal lubricants. The induction of oxidative stress and DNA damage was then assessed at different time points. Our results demonstrate that the paraben mixture was capable of stimulating the generation of mitochondrial and cytosolic reactive oxygen species (ROS), inhibiting sperm motility and viability in a dose-dependent manner. The ability of individual parabens to activate ROS generation and induce oxidative DNA damage was related to alkyl chain length. At the concentration used clinically, methylparaben inhibited sperm motility after both 2 and 5 h exposure (p < 0.05) and affected cell viability (p < 0.01) while augmenting ROS production and oxidative DNA damage. However, DNA fragmentation was not evident following methylparaben exposure. Based on these results, we conclude that, at the concentrations used in commercially available formulations, parabens may impair sperm motility, enhance the generation of mitochondrial ROS and stimulate the formation of oxidative DNA adducts. Taken together, these data underline the potential cytotoxic and genotoxic impact of such compounds in a clinical setting.

Analysis of the effects of polyphenols on human spermatozoa reveals unexpected impacts on mitochondrial membrane potential, oxidative stress and DNA integrity; implications for assisted reproductive technology.

The need to protect human spermatozoa from oxidative stress during assisted reproductive technology, has prompted a detailed analysis of the impacts of phenolic compounds on the functional integrity of these cells. Investigation of 16 individual compounds revealed a surprising variety of negative effects including: (i) a loss of mitochondrial membrane potential (Δψm) via mechanisms that were not related to opening of the permeability transition pore but associated with a reduction in thiol expression, (ii) a decline in intracellular reduced glutathione, (iii) the stimulation of pro-oxidant activity including the induction of ROS generation from mitochondrial and non-mitochondrial sources, (iv) stimulation of lipid peroxidation, (v) the generation of oxidative DNA damage, and (vi) impaired sperm motility. For most of the polyphenolic compounds examined, the loss of motility was gradual and highly correlated with the induction of lipid peroxidation (r=0.889). The exception was gossypol, which induced a rapid loss of motility due to its inherent alkylating activity; one consequence of which was a marked reduction in carboxymethyl lysine expression on the sperm tail; a post-translational modification that is known to play a key role in the regulation of sperm movement. The only polyphenols that did not appear to have adverse effects on spermatozoa were resveratrol, genistein and THP at doses below 100µM. These compounds could, therefore, have some therapeutic potential in a clinical setting.

The intrinsic antinociceptive effects of oxycodone appear to be kappa-opioid receptor mediated.

Our previous studies in the Sprague-Dawley rat showed that the intrinsic antinociceptive effects of oxycodone are naloxone reversible in a manner analogous to morphine but that in contrast to morphine, oxycodone's antinociceptive effects have a rapid onset of maximum effect (approximately 5-7 min compared to 30-45 min for morphine), comprise one antinociceptive phase (compared to two phases) and are of relatively short duration (approximately 90 min compared to approximately 180 min). In the present study, administration of a range of selective opioid receptor antagonists has shown that the intrinsic antinociceptive effects of oxycodone (171 nmol) are not attenuated by i.c.v. administration of (i) naloxonazine, a mu1-selective opioid receptor antagonist, or (ii) naltrindole, a delta-selective opioid receptor antagonist, in doses that completely attenuated the intrinsic antinociceptive effects of equipotent doses of the respective mu- and delta-opioid agonists, morphine and enkephalin-[D-Pen(2,5)] (DPDPE). Although beta-funaltrexamine (beta-FNA) attenuated the antinociceptive effects of oxycodone (171 nmol i.c.v.), it also attenuated the antinociceptive effects of morphine and bremazocine (kappa-opioid agonist) indicative of non-selective antagonism. Importantly, the antinociceptive effects of oxycodone (171 nmol i.c.v.) were markedly attenuated by the prior i.c.v. administration of the selective kappa-opioid receptor antagonist, norbinaltorphimine (nor-BNI), in a dose (0.3 nmol) that did not attenuate the antinociceptive effects of an equipotent dose of i.c.v. morphine (78 nmol). Taken together, these data strongly suggest that the intrinsic antinociceptive effects of oxycodone are mediated by kappa-opioid receptors, in contrast to morphine which interacts primarily with mu-opioid receptors.

The novel N-type calcium channel blocker, AM336, produces potent dose-dependent antinociception after intrathecal dosing in rats and inhibits substance P release in rat spinal cord slices.

N-type calcium channels modulate the release of key pro-nociceptive neurotransmitters such as glutamate and substance P (SP) in the central nervous system. Considerable research interest has focused on the therapeutic potential of the peptidic omega-conopeptides, GVIA and MVIIA as novel analgesic agents, due to their potent inhibition of N-type calcium channels. Recently, the novel peptidic N-type calcium channel blocker, AM336, was isolated from the venom of the cone snail, Conus catus. Thus, the aims of this study were to (i) document the antinociceptive effects of AM336 (also known as CVID) relative to MVIIA following intrathecal (i.t.) bolus dosing in rats with adjuvant-induced chronic inflammatory pain of the right hindpaw and to (ii) quantify the inhibitory effects of AM336 relative to MVIIA on K+-evoked SP release from slices of rat spinal cord. Both AM336 and MVIIA inhibited the K+-evoked release of the pro-nociceptive neurotransmitter, SP, from rat spinal cord slices in a concentration-dependent manner (EC50 values=21.1 and 62.9 nM, respectively), consistent with the antinociceptive actions of omega-conopeptides. Following acute i.t. dosing, AM336 evoked dose-dependent antinociception (ED50 approximately 0.110 nmol) but the doses required to produce side-effects were an order of magnitude larger than the doses required to produce antinociception. For i.t. doses of MVIIA0.07 nmol, produced a dose-dependent decrease in antinociception but the incidence and severity of the side-effects continued to increase for all doses of MVIIA investigated, suggesting that dose-titration with MVIIA in the clinical setting, may be difficult.

Partial trisomy 9--further delineation of the phenotype.

A patient with partial trisomy 9 (47,XX,+9pter----q22.1) had bilateral cleft lip and cleft palate, enophthalmos, severe micrognathia, small, apparently low-set ears, and dislocatable knees. The phenotypic findings are compared with those of other documented cases of total trisomy 9.

Aortitis and large vessel arteritis in a newborn.

Disseminated arteritis with extensive involvement of the aorta, as well as involvement of the arch vessels, coronary arteries, and pulmonary arteries, but not of the arteries within the liver, spleen, kidneys, or other organs, is unusual in a newborn. The presence of both acute and chronic inflammation with fibrosis, as well as calcification and focal ossification in the aorta, would suggest that the process had been present for some time before birth. This lesion should be described rather than designated by an eponym or included with such entities as Takayasu's disease. An additional finding of interest was widespread calcification in Bowman's capsules of numerous glomeruli.