The feeding of heather (Calluna vulgaris) to Teladorsagia circumcincta infected lambs reduces parasitism but can detrimentally impact performance.

Gastrointestinal nematode (GIN) infections impact small ruminant health, welfare, and production across farming systems. Rising anthelmintic resistance and regulation of synthetic drug use in organic farming is driving research and development of sustainable alternatives for GIN control. One alternative is the feeding of plants that contain secondary metabolites (PSMs) e.g., proanthocyanidins (PA, syn. condensed tannins) that have shown anthelmintic potential. However, PSMs can potentially impair performance, arising from reduced palatability and thus intake, digestibility or even toxicity effects. In this study, we tested the trade-off between the antiparasitic and anti-nutritional effects of heather consumption by lambs. The impact of additional feeding of a nematophagous fungus (Duddingtonia flagrans) on larval development was also explored. Lambs infected with Teladorsagia circumcincta or uninfected controls, were offered ad libitum heather, or a control chopped hay for 22 days during the infection patent period. Eight days into the patent period, parasitised lambs were supplemented (or remained unsupplemented) with D. flagrans for a 5-day period. Performance and infection metrics were recorded, and polyphenol levels in the heather and control hay were measured to investigate their association with activity. The lambs consumed heather at approximately 20% of their dry matter intake, which was sufficient to exhibit significant anthelmintic effects via a reduction in total egg output (P = 0.007), compared to hay-fed lambs; the magnitude of the reduction over time in heather fed lambs was almost 10-fold compared to control lambs. Negative effects on production were shown, as heather-fed lambs weighed 6% less than hay-fed lambs (P < 0.001), even though dry matter intake (DMI) of heather increased over time. D. flagrans supplementation lowered larval recovery in the faeces of infected lambs by 31.8% (P = 0.003), although no interactions between feeding heather and D. flagrans were observed (P = 0.337). There was no significant correlation between PA, or other polyphenol subgroups in the diet and egg output, which suggests that any association between heather feeding and anthelmintic effect is not simply and directly attributable to the measured polyphenols. The level of heather intake in this study showed no antagonistic effects on D. flagrans, demonstrating the methods can be used in combination, but provide no additive effect on overall anthelmintic efficacies. In conclusion, heather feeding can assist to reduce egg outputs in infected sheep, but at 20% of DMI negative effects on lamb performance can be expected which may outweigh any antiparasitic benefits.

Outcome of renal transplantation with and without intra-operative diuretics.

AIMS: This paper presents an e-survey of current clinical practice of use of intra-operative diuretics during renal transplantation in the United Kingdom and a study to compare outcome of renal transplants carried out with or without intra-operative diuretics in our centre. METHODS: An e-mail questionnaire to renal transplant surgeons exploring their practice of renal transplantation with or without intra-operative diuretics, the type of a diuretic/s if used and the relevant doses. An observational study comparing the outcome of renal transplant recipients, group no-diuretics (GND, n = 80) carried out from 2004 to 2008 versus group diuretics (GD n = 69) renal transplant recipients who received intra-operative diuretics over a one year period is presented. Outcome measures were incidence of delayed graft function and a comparison of graft survival in both groups. RESULTS: Forty surgeons answered from 18 transplant centres with a response rate of 67%. 13 surgeons do not use diuretics. Mannitol is used by 10/40, Furosemide 6/40 and 11 surgeons use a combination of both. In comparative study there was no significant overall difference in one year graft survival of GD versus GND (N = 65/69, 94% and 75/80, 94% respectively, p = 0.08) and the incidence of delayed graft function was also comparable (16/69, 23% and 21/80, 26% respectively, p = 0.07). The donor characteristics in both groups were comparable. CONCLUSION: The study showed variation in clinical practice on the use of intra-operative diuretics in renal transplantation and it did not demonstrate that the use of diuretics can improve renal graft survival.

Increased expression of keratinase and other peptidases by Candida parapsilosis mutants

Keratinases are enzymes of great importance involved in pathogenic processes of some fungi. They also have a widespread ecological role since they are responsible for the degradation and recycling of keratin. On the one hand, studying them furthers our knowledge of pathogenicity mechanisms, which has important implications for human health, and on the other hand, understanding their ecological role in keratin recycling has biotechnological potential. Here, a wild-type keratinolytic Candida parapsilosis strain isolated from a poultry farm was treated with ethyl methanesulfonate in order to generate mutants with increased keratinase activity. Mutants were then cultured on media with keratin extracted from chicken feathers as the sole source of nitrogen and carbon. Approximately 500 mutants were screened and compared with the described keratinolytic wild type. Three strains, H36, I7 and J5, showed enhanced keratinase activity. The wild-type strain produced 80 U/mL of keratinolytic activity, strain H36 produced 110 U/mL, strain I7, 130 U/mL, and strain J5, 140 U/mL. A 70 percent increase in enzyme activity was recorded for strain J5. Enzymatic activity was evaluated by zymograms with proteic substrates. A peptidase migrating at 100 kDa was detected with keratin, bovine serum albumin and casein. In addition, a peptidase with a molecular mass of 50 kDa was observed with casein in the wild-type strain and in mutants H36 and J5. Gelatinase activity was detected at 60 kDa. A single band of 35 kDa was found in wild-type C. parapsilosis and in mutants with hemoglobin substrate.

Multiple drug resistant Staphylococcus aureus strains isolated from a fish market and from fish handlers

The aim of this study was to investigate the presence of antibiotic resistant Staphylococcus aureus strains in fish stalls and in hands and nasal and oral cavities of fish handlers of the Mucuripe Fish Market, Fortaleza, Ceará, Brazil. All S. aureus isolates were resistant to Ampicillin and 44 percent were multi-drug resistant.

O objetivo da pesquisa foi investigar a presença de Staphylococcus aureus resistente a antibióticos nos boxes de venda de peixe e nas mãos e cavidades nasal e oral de manipuladores de pescado da Feira de Pescado do Mucuripe, Fortaleza, Ceará. Todas as cepas isoladas foram resistentes à ampicilina e 44 por cento apresentaram multiresistência.

Diagnosis and treatment of retropharyngeal injuries in lambs associated with the administration of intraruminal boluses.

The administration of intraruminal identification boluses to a group of 76 lambs resulted in 24 (32 per cent) showing signs of pharyngeal damage. In 16 of them the bolus was found to be in the retropharyngeal region by means of a hand-held microchip scanner, radiography and endoscopy. Purulent tracts were identified in the dorsal pharynx. In 13 of the 16 lambs the surgical removal of the bolus under general anaesthesia was followed by a normal clinical recovery, although the lambs did not grow as well as similar unaffected animals; three of the 24 lambs died. The administration of a larger bolus to 10 yearling rams did not result in any clinical signs of pharyngeal damage. The most important factors influencing the occurrence of this type of injury among this group of lambs were the size and age of the lambs, the positioning of the lambs during bolus administration, the relative size of the dosing gun and bolus, and the large number of animals in the group.

Analysis of a novel strain of murine gammaherpesvirus reveals a genomic locus important for acute pathogenesis.

Infection of mice by murine gammaherpesvirus 68 (MHV-68) is an excellent small-animal model of gammaherpesvirus pathogenesis in a natural host. We have carried out comparative studies of another herpesvirus, murine herpesvirus 76 (MHV-76), which was isolated at the same time as MHV-68 but from a different murid host, the yellow-necked mouse (Apodemus flavicollis). Molecular analyses revealed that the MHV-76 genome is essentially identical to that of MHV-68, except for deletion of 9,538 bp at the left end of the unique region. MHV-76 is therefore a deletion mutant that lacks four genes unique to MHV-68 (M1, M2, M3, and M4) as well as the eight viral tRNA-like genes. Replication of MHV-76 in cell culture was identical to that of MHV-68. However, following infection of mice, MHV-76 was cleared more rapidly from the lungs. In line with this, there was an increased inflammatory response in lungs with MHV-76. Splenomegaly was also significantly reduced following MHV-76 infection, and much less latent MHV-76 was detected in the spleen. Nevertheless, MHV-76 maintained long-term latency in the lungs and spleen. We utilized a cosmid containing the left end of the MHV-68 genome to reinsert the deleted sequence into MHV-76 by recombination in infected cells, and we isolated a rescuant virus designated MHV-76(cA8+)4 which was ostensibly genetically identical to MHV-68. The growth properties of the rescuant in infected mice were identical to those of MHV-68. These results demonstrate that genetic elements at the left end of the unique region of the MHV-68 genome play vital roles in host evasion and are critical to the development of splenic pathology.

The incidence of facial nerve dehiscence at surgery for cholesteatoma.

OBJECTIVE: Facial paralysis can occur after surgery for cholesteatoma. The risk of facial nerve injury is great when the nerve is not covered by its normal bony Fallopian canal. The objective of this study was to identify the incidence of facial nerve dehiscence in patients undergoing surgery for cholesteatoma. STUDY DESIGN: Retrospective chart review. SETTING: Tertiary referral hospital. PATIENT POPULATION: An assessment of all cases performed by the senior author from 1991 to 1999 revealed 59 patients with adequate data available for analysis. These patients ranged in age from 3 to 92 years. In all, 67 surgical procedures. INTERVENTION: Surgery for cholesteatoma, including tympanoplasty and mastoidectomy. MAIN OUTCOME MEASURE: The presence of facial nerve bony dehiscence after exenteration of disease, and postoperative facial nerve function. RESULTS: In 33% of the total procedures analyzed, 30% of the initial procedures, and 35% of the revision procedures, the patients were found to have facial nerve bony dehiscence. The dehiscence was present in the tympanic portion of the facial nerve in the vast majority of patients. Of the 97% of patients with normal preoperative facial nerve function, all retained normal function postoperatively. CONCLUSIONS: Facial nerve dehiscence in our series was far greater than that reported in the literature, underscoring the fact that this is an under-appreciated condition. These findings suggest that surgeons should be highly vigilant when dissecting near the facial nerve. Intraoperative facial nerve monitoring has been shown to be of value in facial nerve preservation during acoustic neuroma resections, and may have a role during surgery for cholesteatoma.

Analysis of the dopamine receptor family in the compact genome of the puffer fish Fugu rubripes.

The genome of the puffer fish, Fugu rubripes (Fugu), is approximately 400 Mb, 7.5 times smaller than that of human. We have isolated four dopamine receptor-like genes from Fugu genomic DNA. These genes show high sequence and structural homology to the known dopamine receptor genes, although, in contrast to previously described genes from this species, the intron size is comparable to that in human. The 5' noncoding exons of the D2/D3 dopamine receptor-like genes is described and compared to that in human. The high gene density of Fugu is shown by the close proximity of a cystatin-like gene 1503 bp from the dopamine receptor gene D222. We propose that the compact genome of Fugu can serve as a model for identifying gene family members directly from genomic DNA.

Relationship between water activity and catalytic activity of lipases in organic media. Effects of supports, loading and enzyme preparation.

The rates of synthesis of dodecyl decanoate in hexane have been measured as a function of water activity (aw), for various immobilised preparations of the lipases from Rhizomucor miehei and Candida rugosa. Only very large changes in the amount of enzyme adsorbed to the support affect the shape of the rate/aw profile; at the highest loadings the profiles tend to become somewhat flatter. A similar levelling can be obtained by pre-adsorbing an inert protein. The effect is probably due to adjacent protein molecules effectively replacing water; it does not simply reflect mass transfer or interfacial area limitation. The activity/aw profile was essentially the same with most supports tested: polypropylene, anion-exchange resin, celite, anion-exchange modified silica. A hydrophobic porous glass support reduced the rate somewhat at intermediate aw values with both enzymes; a polyamide material had this effect only with the lipase from Rh. miehei. The shape of the activity/aw profile was not affected by large differences in purity of the lipase preparation, but did differ between forms that probably differ in glycosylation. Overall, relatively few manipulations of the system can significantly affect the shape of the rate/aw profiles, which seem to be mainly an intrinsic property of the enzyme molecules used.

Lipases from different sources vary widely in dependence of catalytic activity on water activity.

We have measured the rates of esterification in hexane catalysed by suspended immobilised lipases (triacylglycerol acylhydrolase, EC 3.1.1.3), with pre-equilibration to known thermodynamic water activity (a(w)). There were important differences between the enzymes from five different microbes in their retention of activity at low a(w). That from Rhizomucor miehei showed over 40% maximal activity at an a(w) of 0.12, and that from Rhizopus niveus was also fairly active at low a(w). Lipases from other sources required higher a(w) values to show good activity, increasing in the sequence Humicola sp., Candida rugosa and Pseudomonas cepacia. The behaviour was generally similar to two very different support materials, anion-exchange resin and macroporous polypropylene. Comparison of the sequences of the homologous enzymes from Rh. miehei, Rh. niveus and Humicola sp. suggests that changes in charged residues in the 'hinge and lid' region of the structure may be significant in low a(w) tolerance.

Pituitary adenylate cyclase activating polypeptide is a potent vasodilator in humans.

The vasodilator effect of the novel peptide pituitary adenylate cyclase activating polypeptide (PACAP) was investigated in humans. Forearm blood flow was measured in six healthy men by venous occlusion plethysmography. Infusion of PACAP into the brachial artery at 0.01, 0.1, 1, 3, and 10 pmol/min produced a dose-related increase in forearm blood flow in the cannulated arm from 2.8 +/- 0.6 to 8.6 +/- 2.4 ml/100 ml/min at the highest dose (mean +/- SEM, p less than 0.05). In a subsequent experiment, where the highest dose of PACAP was repeated after a 36 min interval, there was no tachyphylaxis of the forearm blood flow response, with the forearm blood flow increasing by 129 +/- 9% during the first infusion and 128 +/- 31% during the second infusion (N.S.). In further experiments, microvascular blood flow was measured by a laser-Doppler flow probe to compare the effects of intradermally injected PACAP, vasoactive intestinal polypeptide (VIP), and calcitonin gene-related peptide (CGRP). When injected into the skin of normal volunteers at 10(-12) to 10(-11) mol/site, each peptide caused a rapid flare lasting 2-3 min, which became erythematous after 5 min. At 10(-12) mol/site, intradermally injected PACAP and VIP caused a maximum increase in skin blood flow at 15 min of 379 +/- 96 and 307 +/- 121% (% increase above basal +/- SEM), respectively, and these responses were not significantly affected by oral aspirin (600 mg) taken 1.5 h beforehand. The vasodilation induced by PACAP at 10(-12) mol/site lasted approximately 6 h, whereas the effect of the same dose of CGRP and VIP lasted less than 2 h. These data suggest that PACAP is a potent and long-lasting vasodilator in humans.

Very high concentrations of islet amyloid polypeptide are necessary to alter the insulin response to intravenous glucose in man.

Islet amyloid polypeptide (IAPP) is a beta-cell peptide that can oppose insulin action in animal systems, but has not been shown to have any action in man; previously, we failed to show an effect of infused IAPP on iv glucose tolerance in human volunteers. We have reexamined its effects at even higher concentrations in six volunteers who received iv glucose (0.5 g/kg) during infusions of IAPP (25 and 50 pmol/kg.min) or normal saline. IAPP rose from a mean basal of 14.7 +/- 5.3 pmol/L to peak levels of 1,420 +/- 110, 2,240 +/- 140, and 27.7 +/- 9 pmol/L, respectively. IAPP at 25 pmol/kg.min had no effect on the plasma glucose disposal rate or the total incremental insulin response, but, in contrast, at 50 pmol/kg.min decreased the insulin response to glucose compared to the saline infusion (incremental area under the curve, 11,276 +/- 2,353 vs. 17,549 +/- 2,687 U; mean +/- SEM; P less than 0.02). This decrease was observed both during the first phase (0-10 min postglucose) insulin response (3,210 +/- 985 vs. 4,382 +/- 815 U; P less than 0.05) and the second phase response (11-90 min, 8,520 +/- 1,719 vs. 13,679 +/- 2,326 U; P less than 0.03). Glucose disposal rate, however, was unaffected (2.0 +/- 0.2 vs. 1.9 +/- 0.2). Thus, circulating IAPP concentrations greater than 90 times normal postprandial peaks were necessary to affect the insulin response to glucose. IAPP appears unlikely to be a circulating hormone influencing carbohydrate metabolism in man.

Should music be used therapeutically in occupational therapy?

It has long been common knowledge that music profoundly affects human beings on a variety of levels. Occupational therapists have at their disposal a potentially powerful therapeutic tool, but the specific effects of this tool have not been documented. Occupational therapists have a history of using music in their treatment, probably to a greater extent than is documented in the literature. It would be advantageous for us to engage in research regarding the application of music to occupational therapy.

Epidemiology of poly-parasitism. III. Effects on the diagnostic capacity of immunological tests.

The paper describes some effects of interaction between different types of parasitic infections on the diagnostic capacity of immunological tests for parasitic diseases. Examples shown include an analysis of association for anergy to tuberculin in skin tests of individuals with and without skin manifestations of onchocerciasis; skin tests with P. westermani and C. sinensis antigens to detect double infections in patients harbouring both trematodes; geographical evaluation of the sensitivity of the complement fixation (CF) test for schistosomiasis with S. mansoni antigen between different communities in Chad; reduced sensitivity of the CF test for schistosomiasis in patients with confirmed schistosomiasis who have also onchocerciasis; studies on the specificity of the slide flocculation (SF) test for schistosomiasis in relation to specified intestinal parasites and to the presence of antibodies to E. granulosus and T. spiralis in villages of Afghanistan; and a study of association between poly-parasitism and anticomplementary activity in the sera of population samples from the African Savannah. The combined investigations indicate that there are large dissimilarities in test performance between communities with different disease spectra and nutritional status. The findings suggest that polyparasitism interferes with immunodiagnostic tests both directly through cross-reactions with antigens and antibodies and indirectly through its effects on nutrition and on mechanisms affecting the complement system. Large scale screening for parasitic diseases with immunodiagnostic tests in areas in which the medical problems are not well-known should be prededed by a small, well-planned pilot study in which the relative sensitivity and specificity of the test can be determined before its use in extensive population studies.

Epidemiology of poly-parasitism. I. Occurrence, frequency and distribution of multiple infections in rural communities in Chad, Peru, Afghanistan, and Zaire.

Parasitic diseases are predominantly rural diseases. They are often associated with poverty, illiteracy, poor sanitation and high risks of exposure to environmental and biological hazards. Because these factors are also essential determinants in the epidemiology of a variety of other infections with quite different etiologies, occurrence of multiple infections in the same people is common. In the tropics, polyparasitism may involve diseases of major public health inportance such as malaria, schistosomiasis, filarial infections, trypanosomiasis, and others. The paper presents data on the frequency and types of multiple infections with different parasitic and other infectious agents for thirteen villages of Chad, Peru and Afghanistan. The age and sex patterns of a number of observed combinations of parasitic and other diseases are shown for different ecological zones. Concomitant infections with up to five species of filarial worms are found in residents of villages in the Congo River Basin of Zaire. The specific types of combinations of these infections vary from place to place and appear to be closely linked to ecological factors.