Effects of freshwater sponge Ephydatia fluviatilis on conjugative transfer of antimicrobial resistance in Enterococcus faecalis strains in aquatic environments.

Filter feeding is a biotic process that brings waterborne bacteria in close contact with each other and may thus support the horizontal transfer of their antimicrobial resistance genes. This laboratory study investigated whether the freshwater sponge Ephydatia fluviatilis supported the transfer of vancomycin resistance between two Enterococcus faecalis strains that we previously demonstrated to exhibit pheromone responsive plasmid conjugation. Microcosm experiments exposed live and dead colonies of laboratory-grown sponges to a vancomycin-resistant donor strain and a rifampicin-resistant recipient strain of Ent. faecalis. Enterococci with both resistance phenotypes were detected on double selection plates. In comparison to controls, abundance of these presumed transconjugants increased significantly in water from sponge microcosms. Homogenized suspensions of sponge cells also yielded presumed transconjugants; however, there was no significant difference between samples from live or dead sponges. Fluorescent in situ hybridization analysis of the sponge cell matrix using species-specific probes revealed the presence of enterococci clusters with cells adjacent to each other. The results demonstrated that sponge colonies can support the horizontal transfer of antimicrobial resistance although the mechanism underlying this process, such as binding of the bacteria to the sponge collagen matrix, has yet to be fully elucidated.

An economic analysis of ransomware and its welfare consequences.

We present in this work an economic analysis of ransomware, a relatively new form of cyber-enabled extortion. We look at how the illegal gains of the criminals will depend on the strategies they use, examining uniform pricing and price discrimination. We also explore the welfare costs to society of such strategies. In addition, we present the results of a pilot survey which demonstrate proof of concept in evaluating the costs of ransomware attacks. We discuss at each stage whether the different strategies we analyse have been encountered already in existing malware, and the likelihood of them being implemented in the future. We hope this work will provide some useful insights for predicting how ransomware may evolve in the future.

Differential expression of syndecans and glypicans in chronically inflamed synovium.

BACKGROUND: Membrane-bound heparan sulphate proteoglycans (HSPGs) act as co-receptors and presenters of cytokines and are involved in cell-matrix and cell-cell adhesion. AIM: To investigate which HSPGs are expressed in knee joint synovia from patients with different forms of arthritis and normal individuals. METHODS: Synovial samples were obtained from patients with early rheumatoid arthritis (n = 8), longstanding rheumatoid arthritis (n = 13), psoriatic arthritis (n = 7), osteoarthritis (n = 6) and normal joints (n = 12). Expression of syndecan-1, -2, -3 and -4 and glypican-1, -3 and -4 was analysed by immunohistochemistry and dual label immunofluorescence. RESULTS: The expression of HSPGs in chronically inflamed synovium exhibited a differential distribution. Syndecan-1 was present in the mononuclear infiltrates of synovia from patients with rheumatoid and psoriatic arthritis where it was expressed by plasma cells. Syndecan-2 was present mainly in blood vessels where it occurred on endothelial cells, pericytes and smooth muscle cells. Syndecan-3 stained intensely in endothelial cells but also occurred in sublining macrophages and the lining layer. Glypican-4 occurred in the lining layer and blood vessels. Increased expression of these HSPGs was apparent in rheumatoid and psoriatic compared to osteoarthritic and normal synovia. Little or no staining for syndecan-4, glypican-1 and glypican-3 was seen in all samples. DISCUSSION: Selected HSPGs, such as syndecan-1, -2 and -3 and glypican-4, could play a part in the pathophysiology of arthritis, such as the migration and retention of leukocytes and angiogenesis in the chronically inflamed synovium.

Guar meal diets as an alternative approach to inducing molt and improving Salmonella enteritidis resistance in late-phase laying hens.

Induced molting of laying hens is a practice used by commercial egg producers to increase the productive lifetime of their flock. However, the conventional method of inducing molt, which involves removal of feed, water, or both as well as a reduction in photoperiod to less than a natural day has drawn criticism due to animal welfare and food safety concerns. The objective of this study was to explore the efficacy of diets containing high levels of guar meal (GM) in inducing molt and reducing susceptibility to Salmonella Enteritidis colonization in late-phase laying hens. Late-phase (68 wk old) Lohmann laying hens were either full-fed standard laying hen diets (nonmolted control), induced to molt by feed withdrawal, or full-fed standard laying hen diets containing 20% GM with or without 250 units/kg of mannanase Hemicell supplementation. On the fourth day of treatment, all hens were orally challenged with SE (1.65 x 10(7) cfu). Hens were killed and evaluated for Salmonella colonization and differences in organ weights 5 d postinoculation. Salmonella Enteritidis present in crop, liver, ovary, and cecal contents were significantly reduced by feeding GM with enzyme supplementation compared with feed withdrawal hens. No significant differences were observed in reproductive tract weights of molted groups, although a difference in liver weight was detected. Results indicate that feeding diets containing 20% GM are as effective as complete feed withdrawal with respect to inducing molt with the added benefit of improved resistance to Salmonella Enteritidis colonization and translocation.

Use of guar by-products in high-production laying hen diets.

A 5x5 Latin square experiment was conducted to evaluate the effect of feeding low concentrations of guar germ or a combination of guar germ and hull (guar meal) in high-production laying hen diets. A total of 125 Lohmann laying hens (21 wk old) of similar BW were randomly assigned to 5 blocks. Each block was divided into 5 experimental units, consisting of 5 hens per unit. Hens were fed either a nonguar control diet, or 1 of 4 diets containing either 2.5 or 5% guar germ, or 2.5 or 5% guar meal over a 20-wk trial period (five 4-wk periods). No significant differences were observed when feeding either 2.5 or 5% guar germ or meal (P>0.05) on hen-day egg production or feed consumption. Significant differences in egg weight, total egg mass per hen, and feed conversion ratio were detected in hens fed 2.5% guar meal, whereas they remained unchanged for diets containing either level of guar germ or 5% guar meal. Feeding either level of guar germ or guar meal did not affect shell quality (shell thickness, egg breaking force, and specific gravity), Haugh units, or egg yolk color (L*, a*, b*). The results showed that both guar germ and guar meal can be fed to high-production laying hens at up to 5% without adverse effects on laying hen performance.

Isolation and characterization of a cytosol protein (64/7.2) present in large amounts in rapidly growing hepatomas.

Protein 64/7.2 (molecular weight/isoelectric point) is present in the cytosol of several hepatomas including the Novikoff hepatoma and Morris hepatomas 9618A, 7794A, and 3924A, but it is not present in liver or 18-hr regenerating liver. Quantitatively, its concentration was highest in Novikoff hepatoma (150 microgram/g tissue) and Morris hepatoma 3924A (550 microgram/g tissue), which are rapid-growing tumors, less in Morris hepatoma 7794A (72 microgram/g tissue), which is of intermediate growth rate, and least in the slow-growing Morris hepatoma 9618A (25 microgram/g tissue). Protein 64/7.2 was isolated from Novikoff hepatoma ascites cells in high purity as shown by its migration as a single band on one-dimensional acid-urea-polyacrylamide gels and a single spot on two-dimensional isoelectric focusing sodium dodecyl sulfate-polyacrylamide gels. Its amino acid composition has an acidic to basic amino acid ratio of 1.6. Its amino-terminal amino acid is lysine, and its carboxyl-terminal amino acid is glycine. Interestingly, the amino acid composition is strikingly similar to that of the phosphorylated Novikoff hepatoma chromatin Protein Cg', partially characterized in our laboratory.

Monoclonal antibody studies suggest a catalytic site at the interface between domains in creatine kinase.

We have located the epitopes recognized by four different monoclonal antibodies which bind to partially unfolded creatine kinase (CK) (ATP: creatine N-phosphotransferase, EC 2.7.3.2) but not to the native enzyme. The epitopes appear to be buried within the CK structure in its native, proteinase-resistant, state. When the epitopes are made accessible to antibody by mild denaturation, CK becomes enzymically-inactive and can be cleaved by proteinase V8 into two large fragments which retain the epitopes and may represent domains. Epitopes on each V8 fragment are associated with highly conserved sequences and are brought physically close to the active site of the enzyme during the later stages of CK refolding and reactivation. The results suggest a catalytic site formed at the interface between two domains which carry the epitopes on their interacting surfaces. Separation of loosely associated domains before or during immunization may account for the origin of antibodies against buried epitopes.

Structural changes in the C-terminal region of human brain creatine kinase studied with monoclonal antibodies.

Epitopes on human brain creatine kinase (B-CK) recognized by three monoclonal antibodies have been located by chemical cleavage methods, followed by peptide synthesis or analysis of specificity for natural variants (isoforms). One antibody, CK-HTB, recognizes a conformational, or assembled, surface epitope on native CK which is also present on partially unfolded forms. It requires an Asn residue at position 300 in the amino acid sequence and will not recognize variants with Lys or His in this position. This results in a striking specificity of the antibody, which binds to B-CK only in chicken and man, but to muscle-form (M-CK) only in the rat. The results suggest that Asn-300 is exposed on the enzyme surface as part of a relatively denaturation-resistant region. Two monoclonal antibodies, CK-END1 and CK-END2, recognise epitopes within 53 amino acids of the C-terminus and bind to a synthetic hexapeptide representing the last six amino acids of human B-CK (Leu-375-Lys-380). The two antibodies show overlapping, but distinct, specificities in their binding to CK variants. CK-END1 requires Met-376 and will not tolerate Ile in this position, whereas CK-END2 requires Leu-375 and will not tolerate Met. Neither antibody binds to native CK, though both will bind to a folding intermediate and to partially unfolded states. This shows that the C-terminus of CK becomes inaccessible to the antibodies during those later stages of protein folding associated with recovery of enzyme activity and suggests that the protein may 'tuck in its tail' during one of the final steps.

Egg storage period and weight effects on hatchability of ostrich (Struthio camelus) eggs.

We studied the affects of storage period and egg weight on the hatchability of 314 ostrich (Struthio camelus) eggs. Eggs were stored at 20 degrees C and 65% RH before incubation at the Poultry Research, Teaching and Extension Center at Texas A&M University (College Station, TX). Eggs were classed by storage period (< or = 5, > 5 < or = 10, > 10 < or = 15, or > 15 < 24 d) and egg weight (< or = 1,450, > 1,450 < or = 1,650, or > 1,650 g) to determine the influence of storage period and egg weight on hatchability, egg weight loss, incubation period, and absolute and relative chick weights. Eggs were incubated at 36.5 to 37.0 degrees C and 25% RH through 38 d of incubation and 36 degrees C and 30% RH thereafter. Mean egg weight loss was greater from eggs of the longest storage period group (> 15 < 24 d) at 21 or 38 d when compared with eggs of the shorter storage periods, but there were no differences at 7, 14, or 28 d among all storage period groups. Mean hatchability was higher in eggs stored < or = 10 d than eggs stored > 15 < 24 d, but hatchability of eggs stored >10 < or = 15 d was not different from eggs stored < or = 10 d or > 15 < 24 d. Incubation period was longer, and absolute and relative weights were higher in eggs stored >15 d than was observed in eggs stored < or = 15 d. Negative correlations were detected between egg weight and moisture loss at 38 d (-0.55) and between hatch time and moisture loss (-0.25). Hatchability was higher in small eggs than medium eggs (< or = 1,650 g). A positive correlation was observed between chick and egg weights (0.84). The results indicated that storage period and egg weight affected egg weight loss. Our results suggested that the most effective storage period was less than 15 d to maintain hatchability for ostrich eggs when incubated at 36.5 to 37.0 degrees C with 25% RH.

Effects of guar meal by-product with and without beta-mannanase Hemicell on broiler performance.

Guar gum production yields a high protein guar meal that can be subdivided into germ and hull fractions. Feeding high concentrations of guar meal reduces body weight and feed efficiency in chickens due to the presence of a residual guar gum. Two experiments determined the upper feeding levels of guar meal and the hull and germ fractions in broiler chickens. An industrial source beta-mannanase (Hemicell) also was fed in combination with guar meals. Experiment 1 utilized a 3 x 4 factorial design to feed broiler chickens diets containing guar germ, guar hull, or guar meal at 4 levels (2.5, 5.0, 7.5, and 10.0%) compared with a negative control diet. Results indicated that any of the 3 guar meals could be fed at a 2.5% dietary inclusion rate without adversely affecting broiler chicken growth to 6 wk of age. In experiment 2, a 4 x 2 factorial design consisting of the 3 by-products meals at 5% inclusion and soybean meal control with and without enzyme tested whether Hemicell could increase inclusion rates without decreasing broiler growth or feed consumption to 6 wk of age. Addition of Hemicell to feed had no effect on measures of growth in chickens fed the control diet. Hemicell significantly improved feed:gain ratio of diets containing 5% of each fraction of guar meal versus the untreated diets. Feed:gain ratio for the Hemicell-treated 5% germ fraction diet was improved to control diet levels. Results indicated that the upper feeding level of guar meal and germ and hull fraction of guar meal is 2.5%, and addition of beta-mannanase (Hemicell) increases the upper feeding level for the germ fraction to 5%.

Monoclonal antibodies against defined regions of the muscular dystrophy protein, dystrophin.

Nineteen monoclonal antibodies which bind to native dystrophin in the plasma membrane of frozen muscle sections were obtained using a recombinant fusion protein as immunogen. On Western blots of normal mouse muscle extracts, the antibodies bind specifically to a 400,000 Mr protein which is absent from dystrophic mouse (mdx) muscle. At least four distinct epitopes have been identified by cleavage mapping methods. Although the fusion protein contained 25% of the human dystrophin sequence (Cys816-Asp1747; Mr 108,000), most of the monoclonal antibodies (15 out of 19) recognize a single fragment of Mr 27,500.

Monoclonal antibody evidence for structural similarities between the central rod regions of actinin and dystrophin.

A monoclonal antibody, MANDYS141, binds to both dystrophin and actinin on Western blots (SDS-denatured), but only to actinin in frozen sections of human muscle (native conformation). It differs from a polyclonal cross-reacting antiserum in that it binds to several muscle isoforms of actinin (smooth, fast and slow) from man, mouse and chicken and recognises a quite different part of the proposed triple-helical region of dystrophin (amino acids 1750-2248). The results suggest that structural homologies between actinin and dystrophin occur more than once in their central helical regions and provide experimental support for an actinin-like central rod model for dystrophin.

Treatment of human muscle creatine kinase with glutaraldehyde preferentially increases the immunogenicity of the native conformation and permits production of high-affinity monoclonal antibodies which recognize two distinct surface epitopes.

Treatment of human muscle creatine kinase (MM-CK) with glutaraldehyde produced highly aggregated forms which retained the native antigenic structure. Immunization of BALB/c mice with CK aggregates instead of untreated CK produced over ten-fold higher titres of antibody against native CK without increasing the titres of antibody against denatured enzyme. Production of high-affinity monoclonal antibodies specific for both the muscle isoenzyme and the native conformation became possible where the use of untreated CK had failed. Four monoclonal antibodies have been characterized by an epitope mapping technique and compared with a commercially available monoclonal antibody. One antibody has a much higher affinity for MM-CK than the other three and the commercial antibody. Competition studies show that it also recognizes a different epitope on the CK surface from the other three monoclonal antibodies which bind to the same surface region as the commercial antibody. Immunoassays based on the high affinity antibody can easily measure less than 1 ng of CK, a sensitivity comparable to, or better than, standard enzymatic assays.

Review article: Oral, modified-release mesalazine formulations--proprietary versus generic.

Products containing mesalazine have been used in the treatment of inflammatory bowel disease for many years. Many of the oral, modified-release products are reaching the point of patent expiration, and it is expected that several new 'generic' versions will be developed. As mesalazine acts topically, the drug needs to be available at the site of inflammation to be effective. For this reason, the currently available products have been developed with individual formulations so that physicians have a choice when matching the different release profiles to the site and extent of disease. As such, the current guidelines state that oral, delayed-release mesalazine formulations are not interchangeable and should be prescribed by their proprietary (brand) name. The standard regulatory assessment process for generic or 'copy' products, using systemic bioequivalence data, does not appear to be sufficient when evaluating topically acting, oral, modified-release products. We therefore recommend that the regulatory bodies should require that new, oral mesalazine products should be assessed by a combination of dissolution, bioequivalence and (a minimum of one) adequately powered, comparative trial to determine therapeutic equivalence. Of most importance here is that the assessment of new modified-release products is sufficiently rigorous to allow patients and physicians to be confident in their use.

The planes of the utricular and saccular maculae of the guinea pig.

To establish a link between otolith anatomy and function it is necessary to know the regions of the utricular and saccular maculae, which are stimulated by any arbitrary linear acceleration stimulus. That requires accurate information about the location and orientation of the spatially extended maculae in head-fixed coordinates and referred to head-fixed landmarks (such as Reid's line). New data showing the location of the otolithic maculae in the guinea pig with respect to head-fixed stereotaxic coordinates are presented. Guinea pigs were perfused with Karnovsky's fixative and the maculae were exposed while the head was held in a guinea pig stereotaxic device. An electrolytically sharpened fine wire held in a calibrated micromanipulator was touched to points all over the surface of each macula under visual observation with the aid of a high-power operating microscope. The x, y, z coordinates of these points were plotted using a three-dimensional plotting program. Both maculae have pronounced curvature so that dorsoventral shear forces will stimulate regions of both the utricular and saccular maculae.

Semicircular canal occlusion causes permanent VOR changes.

We measured the guinea pig horizontal vestibulo-ocular reflex (hVOR) to high acceleration impulsive head rotations following a unilateral lateral semicircular canal (LSCC) occlusion. We found a significant hVOR deficit for rotations toward the side of the occluded LSCC and this deficit did not show systematic changes over 3 months. We considered the LSCC nerve was still functional as shown by the normal appearance of the crista of the LSCC ampulla and also electrical stimulation of the LSCC. We conclude that the VOR during angular acceleration in response to high acceleration shows no adaptive plasticity following a unilateral LSCC occlusion.

Suicidal nonfatal impalement injury of the thorax.

We treated an impalement injury of the thorax resulting from a suicide attempt in the form of a road traffic accident. The patient survived and was discharged 5 weeks after his injury. The surgical management of thoracic impalement injuries and the rationale behind a multidisciplinary approach are discussed.

Social class differences in health and care in the year before death.

STUDY OBJECTIVE: The aim was to identify any social class differences in health and care in the year before death emerging from a wider study of life before death. DESIGN AND SETTING: Data were collected at interviews, mainly of close relatives, with those who knew most about the people's lives in the year before they died. The sample was a random sample of adult (15 and over) deaths in 10 areas of England in 1987. SUBJECTS: Information was obtained about 639 persons, 80% of the initial sample of 800 deaths. MAIN RESULTS: While middle class people die at an older age, the symptoms and physical restrictions reported for middle class and working class people were similar, and middle class people were reported to have a better quality of life before death. More working class people were felt to be in financial need. CONCLUSIONS: Money and class contribute to the quality of life before death as well as postponing death.

Why don't more young men in the UK become fathers?

OBJECTIVE: This paper aims to show that, compared with young women, there is an apparent discrepancy in the reported sexual behaviour of young men and records of their fatherhood. DESIGN: The data come from four studies of the sexual behaviour of young people in various parts of Britain between 1960 and 1990 and from statistics published by the Office of Population Censuses and Surveys. MAIN RESULTS: More of the young men than the young women had been sexually active, and the men had had more sexual partners. Relatively few men under 20, however, are identified as fathers. In 1991 there were 52,396 live births to women under 20 years, but only 12,959 births were attributed to men under 20, and if all those to women under 20 for which the father was not identified are added to this, the total is still only 28,208--54% of the number of births to women of that age. Seven possible reasons for the discrepancy are discussed. Data suggest that the two most probable explanations are that there are different patterns of intercourse among young men and women and that the data obtained in surveys are to some extent inaccurate or biased. CONCLUSIONS: Differing behaviours indicate a double standard for men and women. Imperfect information affects predictions about a potential heterosexual HIV epidemic.