Impact of artificial accelerated ageing of PVC surfaces and surface degradation on disinfectant efficacy.

BACKGROUND: Standardized efficacy surface tests for disinfectants are performed on pristine surfaces. There is a growing interest in understanding the impact of surface ageing on disinfectant activity, owing for example to the increased usage of ultraviolet (UV) radiation and oxidative chemistries for surface decontamination. This acknowledges that general surface 'wear and tear' following UV radiation and oxidative biocide exposure may impact biocidal product efficacy. METHODS: PVC surfaces were aged through thermal and UV-A radiation (340 nm wavelength) following the use of standard ageing surface protocols to simulate natural surface degradation. Surface roughness, contact angle and scanning electron microscopy were performed to evaluate physical changes in PVC surfaces before and after artificial ageing. The efficacy of five pre-impregnated disinfectant wipes were evaluated using the ASTM E2967-15 on stainless-steel (control) and PVC surfaces (aged and non-aged). RESULTS: The type of formulation and the organism tested remained the most significant factors impacting disinfectant efficacy, compared with surface type. Both thermal ageing and UV-A exposure of PVC surfaces clearly showed signs of surface degradation, notably an increase in surface roughness. Physical changes were observed in the roughness of PVC after artificial ageing. A difference in disinfectant efficacy dependent on aged PVC surfaces was observed for some, but not all formulations. CONCLUSION: We showed that surface type and surface ageing can affect biocidal product efficacy, although in a non-predictable manner. More research is needed in this field to ascertain whether surface types and aged surfaces should be used in standardized efficacy testing.

Evaluating the antimicrobial efficacy of long-lasting hand sanitizers on skin.

BACKGROUND: The microbicidal efficacy of hand sanitizer formulations is usually measured through standardized quantitative suspension tests and fingerpad tests; these cannot evaluate long-lasting formulations or are impractical due to biological risks, high cost, or time required for testing. With increased numbers of long-lasting microbicidal activity claims of commercially available hand sanitizers, alternative testing strategies are required. AIM: To explore the use of a standardized ex-vivo pig skin model to reproducibly measure long-lasting efficacy of an alcohol-free hand sanitizer formulation. METHODS: The microbicidal efficacy of an alcohol-free hand sanitizer was tested against Staphylococcus aureus, Escherichia coli and Klebsiella pneumoniae, and the enveloped virus SARS-CoV-2 with quantitative suspension tests (EN13727 and EN14476) with a contact time of 5 min. The product was then tested over a 6 h period using an ex-vivo pig skin model with a modified version of PAS 2424 to simulate the impact of skin abrasion. FINDINGS: Quantitative suspension tests yielded a >5 log10 reduction for all organisms tested within a 5 min contact time. Pig skin tests showed reduced but consistent efficacy at all time points and indicated no significant impact of abrasion on efficacy. CONCLUSION: The use of the ex-vivo pig skin model provides a potentially viable and convenient model system to test long-lasting hand sanitizer formulations, providing a path for sustainable hand sanitizer formulation claims of activity on skin.

The 2010 Royal Australasian College of Physicians' policy statement 'Circumcision of infant males' is not evidence based.

Infant male circumcision (MC) is an important issue guided by Royal Australasian College of Physicians (RACP) policy. Here we analytically review the RACP's 2010 policy statement 'Circumcision of infant males'. Comprehensive evaluation in the context of published research was used. We find that the Statement is not a fair and balanced representation of the literature on MC. It ignores, downplays, obfuscates or misrepresents the considerable evidence attesting to the strong protection MC affords against childhood urinary tract infections, sexually transmitted infections (human immunodeficiency virus, human papilloma virus, herpes simplex virus type 2, trichomonas and genital ulcer disease), thrush, inferior penile hygiene, phimosis, balanoposthitis and penile cancer, and in women protection against human papilloma virus, herpes simplex virus type 2, bacterial vaginosis and cervical cancer. The Statement exaggerates the complication rate. Assertions that 'the foreskin has a functional role' and 'is a primary sensory part of the penis' are not supported by research, including randomised controlled trials. Instead of citing these and meta-analyses, the Statement selectively cites poor quality studies. Its claim, without support from a literature-based risk-benefit analysis, that the currently available evidence does 'not warrant routine infant circumcision in Australia and New Zealand' is misleading. The Statement fails to explain that performing MC in the neonatal period using local anaesthesia maximises benefits, safety, convenience and cost savings. Because the RACP's policy statement is not a fair and balanced representation of the current literature, it should not be used to guide policy. In the interests of public health and individual well-being, an extensive, comprehensive, balanced review of the scientific literature and a risk-benefit analysis should be conducted to formulate policy.

Relative efficacies of omega-3 polyunsaturated fatty acids in reducing expression of key proteins in a model system for studying osteoarthritis.

OBJECTIVE: To assess the relative efficacy of three different omega-3 (n-3) polyunsaturated fatty acids (PUFAs) in suppressing the mRNA levels for important proteins involved in the etiology of osteoarthritis (OA). METHODS: A model cell culture system (bovine chondrocytes) was used. Inflammatory factors and enzymes involved in OA were induced by exposure of the chondrocyte cultures to interleukin-1alpha (IL-1alpha). The effect of pre-incubating cultures with various amounts of exogenous fatty acids on subsequent levels of mRNAs was assessed by reverse transcription-polymerase chain reactions (RT-PCR). RESULTS: Exposure of cultures to IL-1alpha induced expression of the cartilage proteinases A Disintegrin And Metalloproteinase with ThromboSpondin motifs (ADAMTS)-4 and ADAMTS-5, cyclooxygenase (COX)-2, the matrix metalloproteinase (MMP)-3 and the inflammatory cytokines IL-1alpha, interleukin-1beta (IL-1beta) and tumour necrosis factor-alpha (TNF-alpha). n-3 PUFAs were able to reduce the levels of mRNA for ADAMTS-4, ADAMTS-5, MMP-3, MMP-13, COX-2 (but not COX-1), IL-1alpha, IL-1beta and TNF-alpha. Eicosapentaenoic acid (EPA) was the most effective, followed by docosahexaenoic (DHA) and then alpha-linolenic (ALA) acid. The n-6 PUFA, arachidonic acid (AA) had no effect. CONCLUSION: These results show that omega-3 (n-3) PUFAs cause a reduction in the mRNA levels for various proteins known to be important in the pathology of OA. They provide a molecular explanation, at least in part, for beneficial effects of dietary omega-3 PUFAs for the amelioration of symptoms of the disease. The relative efficacy of EPA suggests that this omega-3 PUFA may be especially useful for dietary supplementation in patients with OA.

Adhesive multiplicity in the interaction of embryonic fibroblasts and myoblasts with extracellular matrices.

Neff et al. (1982, J. Cell Biol., 95:654-666) have described a monoclonal antibody, CSAT, directed against a cell surface antigen that participates in the adhesion of skeletal muscle to extracellular matrices. We used the same antibody to compare and parse the determinants of adhesion and morphology on myogenic and fibrogenic cells. We report here that the antigen is present on skeletal and cardiac muscle and on tendon, skeletal, dermal, and cardiac fibroblasts; however, its contribution to their morphology and adhesion is different. The antibody produces large alterations in the morphology and adhesion of skeletal myoblasts and tendon fibroblasts; in contrast, its effects on the cardiac fibroblasts are not readily detected. The effects of CSAT on the other cell types, i.e., dermal and skeletal fibroblasts, cardiac muscle, 5-bromodeoxyuridine-treated skeletal muscle, lie between these extremes. The effects of CSAT on the skeletal myoblasts depends on the calcium concentration in the growth medium and on the culture age. We interpret these differential responses to CSAT as revealing differences in the adhesion of the various cells to extracellular matrices. This interpretation is supported by parallel studies using quantitative assays of cell-matrix adhesion. The likely origin of these adhesive differences is the progressive display of different kinds of adhesion-related molecules and their organizational complexes on increasingly adhesive cells. The antigen to which CSAT is directed is present on all of the above cells and thus appears to be a lowest common denominator of their adhesion to extracellular matrices.

The cell substrate attachment (CSAT) antigen has properties of a receptor for laminin and fibronectin.

The cell substrate attachment (CSAT) antigen is an integral membrane glycoprotein complex that participates in the adhesion of cells to extracellular molecules. The CSAT monoclonal antibody, directed against this complex, inhibited adhesion of cardiac and tendon fibroblasts and skeletal myoblasts to both laminin and fibronectin, thus implicating the CSAT antigen in adhesion to these extracellular molecules. Equilibrium gel filtration was used to explore the hypothesis that the CSAT antigen functions as a cell surface receptor for both laminin and fibronectin. In this technique, designed for rapidly exchanging equilibria, the gel filtration column is pre-equilibrated with extracellular ligand to ensure receptor occupancy during its journey through the column. Both laminin and fibronectin formed complexes with the CSAT antigen. The association with laminin was inhibited by the CSAT monoclonal antibody; the associations with both fibronectin and laminin were inhibited by synthetic peptides containing the fibronectin cell-binding sequence. Estimates of the dissociation constants by equilibrium gel filtration agree well with those available from other measurements. This suggests that these associations are biologically significant. SDS PAGE showed that all three glycoproteins comprising the CSAT antigen were present in the antigen-ligand complexes. Gel filtration and velocity sedimentation were used to show that the three bands comprise and oligomeric complex, which provides an explanation for their functional association. The inhibition of adhesion by the CSAT monoclonal antibody and the association of the purified antigen with extracellular ligands are interpreted as strongly implicating the CSAT antigen as a receptor for both fibronectin and laminin and perhaps for other extracellular molecules as well.

Integrin (the CSAT antigen): functionality requires oligomeric integrity.

Integrin, the cell-substrate attachment (CSAT) antigen, is a complex of integral membrane glycoproteins whose apparent function is to mediate cell-substratum adhesion by serving as a transmembrane link between the extracellular matrix and elements of the cytoskeleton. Previous attempts to separate the members of this complex under nondenaturing conditions have been successful. We have now produced a monoclonal antibody "G" that is specific for the lower molecular mass cysteine-rich band 3 of the complex. Using an antibody affinity column containing this monoclonal antibody, it is possible to dissociate integrin into two fractions, one containing band 3, the other containing bands 1 plus 2. Neither fraction will by itself bind fibronectin, laminin, or talin. However, when the fractions are combined, the reconstituted integrin elutes from a gel filtration column in the same position as the native complex, and binding activity to these molecules returns. Further, it is shown by gel filtration that the recognition site for the adhesion-disrupting monoclonal antibodies CSAT and JG22 is on band 3, supporting the contention that integrin is an oligomer. The data presented here is consistent with integrin being either a mixture of heterodimers, each with a common subunit and reacting with a particular extracellular matrix molecule, or a single heterotrimer capable of binding to several different extracellular matrix molecules.

Application of Guideline Recommended Treatment in Routine Clinical Practice: A Population-based Study of Stage I-IIIB Non-small Cell Lung Cancer.

AIMS: The application of guideline recommended treatment (GRT) in routine clinical practice can be difficult due to differences between the clinic population and the clinical trial populations on which evidence is based. The study aims were to measure receipt of GRT in stage I-IIIB non-small cell lung cancer (NSCLC) patients, identify factors associated with GRT and its impact on survival. MATERIALS AND METHODS: New diagnoses of stage I-IIIB NSCLC from 1 January 2006 to 31 December 2011 in South West Sydney residents were identified from the district Clinical Cancer Registry. Treatment received was assigned as GRT or not based on Australian guidelines (using Eastern Cooperative Oncology Group [ECOG] performance status and TNM stage). Multivariate Poisson regression models with robust variance identified predictors of GRT receipt. Cox regression models identified multivariate predictors of patient survival. RESULTS: In total, 592 eligible cases were identified, of whom 66% (n = 389) received GRT. This ranged from 81% of stage I to 39% of stage IIIB (relative risk 0.48, 0.38-0.60, P < 0.0001). Stage I-IIIA patients who were ECOG 2 and stage III patients aged 70 years and older were less likely to receive GRT. The median survival was 30 months in the GRT group and 16 months in the non-GRT group (P < 0.001). GRT receipt was associated with improved survival in stage I-II disease only (hazard ratio 0.41, P < 0.001; and hazard ratio 0.43, P = 0.006). CONCLUSION: One-third of NSCLC patients did not receive GRT. Stage and performance status were key predictors for GRT receipt. Patients with early stage NSCLC were associated with improved survival with the receipt of GRT.

Integrins isolated from Rous sarcoma virus-transformed chicken embryo fibroblasts.

Avian integrins are a complex of three integral membrane glycoproteins that are thought to have a role both in anchoring the cytoskeleton to the plasma membrane and establishing linkages to the extracellular matrix. We previously demonstrated that bands 2 and 3 of integrin are phosphorylated on both tyrosine and serine residues in chicken embryo fibroblasts (CEF) transformed with Rous Sarcoma virus (RSV) and other oncogenic retroviruses. The effects of RSV transformation on integrins from chick cells are now further characterized. The major site of tyrosine phosphorylation on band 3 in RSV transformed CEF has been identified as tyrosine 788. We also demonstrate that the product of the RSV oncogene, pp60v-src, can phosphorylate integrin in vitro, at the same residue. Tryptic peptide mapping and tunicamycin treatment indicates that a previously observed 4-8 k increase in the Mr of integrins from RSV-transformed cells can be attributed to an alteration in a post-translational modification such as glycosylation. Equilibrium gel filtration assays were used to test the ability of integrins from RSV-transformed CEF to interact with talin and fibronectin. Tyrosine phosphorylated integrins showed a decreased ability to interact with both these ligands in vitro. Conversely, integrins isolated from RSV-transformed CEF metabolically labeled in the absence of phosphatase inhibitor contained only low levels of phosphotyrosine and showed an almost normal ability to interact with ligands. Competition experiments indicated that the region of integrin containing tyrosine 788 is also important for talin binding to integrins.

The effect of dietary sodium on the concentrations of vasoactive intestinal peptide in plasma and lung.

STUDY OBJECTIVES: In this study, we sought to determine whether changes in the concentration of vasoactive intestinal peptide (VIP) in the lung might explain the increase in bronchial reactivity associated with high sodium diets. DESIGN: Male Sprague-Dawley rats, eight in each group, were placed on low-sodium, normal-sodium, or high-sodium diets and distilled drinking water ad libitum for 7 days. On the day of study, blood was sampled to determine plasma VIP concentration and the lungs were harvested and snap frozen in liquid nitrogen. VIP was measured in plasma and tissue extracts by radioimmunoassay. RESULTS: The VIP concentrations in both lung and plasma varied with dietary sodium. Plasma VIP level was significantly higher in the rats that had received the low-sodium diet (51.45 +/- 7.35 pmol L-1) than in the rats that had received the high-sodium diet (29.84 +/- 6.83; p < 0.05). In the lung, VIP level was greater in the rats that had received the normal-sodium diet (378.13 +/- 41.68 fmol/g) than in rats that had received either the low-sodium diet (137.30 +/- 26.11 fmol/g; p < 0.0005) or the high-sodium diet (182.64 +/- 28.63 fmol/g; p < 0.005). CONCLUSIONS: The lower plasma and pulmonary concentrations of VIP observed in rats that had received a high-sodium diet suggest that VIP may play a role in the increased bronchial reactivity reported with this diet.

Accuracy of single concentration estimations of platelet angiotensin II receptor number. Its usefulness in screening for pregnancy-induced hypertension.

Platelet angiotensin II (AngII) receptor number has been suggested as a screening test for pregnancy-induced hypertension. However, markedly different false-positive rates have been reported, perhaps the result of differing methods used. We sought therefore to compare the two methods. Platelet AngII receptor number was determined by saturation analysis with computerized curve fitting and specific binding at a single radioligand concentration. The two methods were compared by correlation and by plotting their differences v their means, to determine their limits of agreement. There were significant correlations between the value obtained by saturation analysis and each of the three single ligand concentrations studied (1 nmol/L, P < .001; 500 pmol/L, P < .001; and 250 pmol/L, P < .01). However, for none of the three did the regression line approach the line of equality. Assessment of agreement by comparing differences and means for each subject showed increasing scatter with increasing receptor number and 95% confidence intervals too large to be clinically relevant. We conclude that the receptor number estimated from specific binding at one ligand concentration differs significantly from that obtained by saturation analysis. The limits of agreement of the two methods are wide and we urge caution in the use of single ligand concentration methods for estimating binding site densities.

Angiotensin-converting enzyme inhibition with enalapril increases the cardiac concentration of vasoactive intestinal peptide.

In patients with congestive cardiac failure, treatment with ACE inhibitors results in peripheral vasodilatation and an increase in cardiac output without an increase in heart rate, which suggests a positive inotropic effect. This cannot be explained by the changes in angiotensin II and bradykinin concentrations that occur. It has been suggested that ACE also metabolizes VIP, which is a positive inotrope. As VIP is synthetized by the heart and acts locally to increase cardiac output, we postulated that ACE inhibition would increase the myocardial concentration of VIP. Male Sprague-Dawley rats received enalapril (2 mg/kg/day) in the drinking water or no therapy for seven days. On day seven they were anaesthetized and blood sampled. The hearts and kidneys were then harvested and snap frozen by immersion in liquid nitrogen. Concentrations of VIP in plasma and tissue extracts were measured by radioimmunoassay. Plasma and renal concentrations of VIP did not change in the enalapril-treated rats. However, the myocardial concentration of VIP increased significantly in the rats receiving enalapril compared with control animals (p < 0.0005). We conclude that treatment with ACE inhibitors results in increased myocardial VIP concentrations and suggest that this may contribute to the improvement in cardiac function that occurs with these agents.

Effects of enalapril on vasoactive intestinal peptide metabolism and tissue levels.

Angiotensin converting enzyme inhibitor therapy results in an increase in cardiac output without an increase in heart rate suggesting a positive inotropic effect. This cannot be explained by changes in angiotensin II and bradykinin concentrations. Angiotensin converting enzyme may also metabolise vasoactive intestinal peptide (VIP), a vasodilator and positive inotrope whose concentration in the heart declines in heart failure. We sought to determine whether changes in plasma VIP or its metabolism might explain the positive inotropic effect of angiotensin converting enzyme inhibitors. We also measured VIP in the heart to determine whether a local increase in VIP might explain this effect. Male Sprague-Dawley rats were randomised to control and enalapril groups (2 mg kg(-1) day(-1)). After 7 days, rats were anaesthetised and underwent metabolic clearance studies for VIP or had hearts, lungs and kidneys removed and snap frozen. VIP concentrations in plasma, infusate and tissue extracts were measured by radioimmunoassay. Plasma concentrations of VIP were unchanged by treatment with enalapril (control: 7.7 +/- 0.8 pmol l(-1); enalapril: 7.9 +/- 0.8 pmol l(-1) ), while the metabolic clearance rate of) VIP increased significantly (control: 10.4 +/- 1.4 ml min(-1) 100 g(-1); enalapril: 17.3 +/- 1.6 ml min(-1) 100 g(-1); p < 0.005). Secretion rate) also increased in enalapril treated rats (139.1 +/- 25.0 pmol min(-1) 100 g(-1) compared with controls (96.3 +/- 13.4 pmol min (-1) 100 g(-1); P< 0.01). VIP in the heart increased after enalapril (control: 208.4 +/- 39.0 pmol g (-1); enalapril: 928.9 +/- 123.6 fmol g(-1); P < 0.0005). Angiotensin converting enzyme inhibition increases the metabolism of VIP. However, the significant increase in the myocardial concentration of VIP may contribute to the beneficial haemodynamic inotrope effects of angiotensin converting enzyme inhibitors.

Effect of alcohol content on beer consumption by rats.

Three brands of beer of varying alcohol content were presented to separate groups of male rats. The consumption data indicated, firstly, an inverse relationship between acceptability of the beer and its alcohol content. The data also indicated that the acceptability of full-strength beer could be augmented by prior exposure to low-alcohol and alcohol-free beer. The results are discussed with reference to orosensory- and inebriation-based models of alcohol consumption.

Verapamil prevents post-transplant oliguric renal failure.

Verapamil has proven effective in preventing acute renal failure in animal models if given prior to the insult and hence possibly has a role in the preservation of cadaveric renal tissue for transplantation. Twenty renal donors were randomly assigned to treatment (receiving verapamil 20 mg intravenously) and control groups. Recipients were monitored for renal failure by urine output and serum creatinines on days 1 and 7 and dialysis requirement to one week. Early urine outputs and serum creatinines (day 1) were significantly better in the treated than control group (p greater than 0.01, 0.05 respectively). We conclude therefore that verapamil may prevent post-transplant acute renal failure, but its optimal dosage and route of administration remain to be determined.