A serologic study of canine herpes virus-1 infection in the Norwegian adult dog population.

Canine herpes virus-1 (CHV1) causes a fatal hemorrhagic disease in neonatal puppies and is associated with reproductive problems in female dogs. This serologic study was conducted to assess the seroprevalence of CHV1 infection in Norway. Blood samples were collected from clinically healthy dogs (n = 436) one yr of age and older of both genders, supplied by four small animal clinics (A, B, C and D) in different parts of the country. The immunoperoxidase monolayer assay was used for testing of CHV1 antibodies. Serum titers were recorded as the reciprocal value of the highest dilution producing specific cell staining. Titers equal to or above 80 were considered positive for exposure to CHV1. In total, 80.0% of the dogs had titers ≥80 and were classified as positive. Mean age for seronegative dogs was 4.7 yrs (95% CI 4.1-5.4) and for seropositive dogs 5.0 yrs (95% CI 4.7-5.4). Of the dogs, 32.8% displayed a weakly positive titer of 80, whereas 41.5 and 5.7% fell into the moderately (titer 160 and 320) and strongly (titer ≥640) positive categories, respectively. No association was demonstrated when comparing CHV1 antibody titers to gender or reproductive parameters like previous matings, pregnancies, births or number of puppies born. Age, visit in foreign countries and clinic explained together 78% of the variation in antibody titer categories. The percentage of positive samples differed significantly between the four clinics (A 98%, B 58.5%, C 74.6%, D 89.5%). A reasonable explanation for this finding has not been established. No information about an ongoing outbreak of CHV1 infection was available. In conclusion, this study strongly indicates that CHV1 infection is endemic in the dog population of Norway. There are significant differences in seroprevalence between geographic regions in the country.

Retracted: 147 reduced syntaxin-5 in skeletal muscle of patients with type 2 diabetes. A link between lipid storage and insulin resistance.

This article has been retracted: please see Elsevier Policy on Article Withdrawal (http://www.elsevier.com/locate/withdrawalpolicy). This abstract has been retracted at the request of Jan Borén, co-author, because of conscious fabrication, corruption or suppression of basic material and conscious preparation and presentation of falsified results in the abstract by one of the authors.

Re-targeted adenovirus vectors with dual specificity; binding specificities conferred by two different Affibody molecules in the fiber.

Vectors based on Adenovirus type 5 (Ad5) are among the most common vectors in cancer gene therapy trials to date. However, for increased efficiency and safety, Ad5 should be de-targeted from its native receptors and re-targeted to a tumor antigen. We have described earlier an Ad5 vector genetically re-targeted to the tumor antigen HER2/neu by a dimeric version of the Affibody molecule ZH inserted in the HI-loop of the fiber knob of a coxsackie and adenovirus receptor-binding ablated fiber. This virus showed almost wild-type growth characteristics and infected cells through HER2/neu. Here we generate vectors with double specificity by incorporating two different Affibody molecules, ZH (HER2/neu-binding) and ZT (Taq polymerase-binding), at different positions relative to one another in the HI-loop. Receptor-binding studies together with viral production and gene transfer assays showed that the recombinant fiber with ZT in the first position and ZH in the second position (ZTZH) bound to both its targets, whereas surprisingly, the fiber with ZHZT was devoid of binding to HER2/neu. Hence, it is possible to construct a recombinant adenovirus with dual specificity after evaluating the best position for each ligand in the fiber knob.

Adenovirus 5 vector genetically re-targeted by an Affibody molecule with specificity for tumor antigen HER2/neu.

In order to use adenovirus (Ad) type 5 (Ad5) for cancer gene therapy, Ad needs to be de-targeted from its native receptors and re-targeted to a tumor antigen. A limiting factor for this has been to find a ligand that (i) binds a relevant target, (ii) is able to fold correctly in the reducing environment of the cytoplasm and (iii) when incorporated at an optimal position on the virion results in a virus with a low physical particle to plaque-forming units ratio to diminish the viral load to be administered to a future patient. Here, we present a solution to these problems by producing a genetically re-targeted Ad with a tandem repeat of the HER2/neu reactive Affibody molecule (ZH) in the HI-loop of a Coxsackie B virus and Ad receptor (CAR) binding ablated fiber genetically modified to contain sequences for flexible linkers between the ZH and the knob sequences. ZH is an Affibody molecule specific for the extracellular domain of human epidermal growth factor receptor 2 (HER2/neu) that is overexpressed in inter alia breast and ovarian carcinomas. The virus presented here exhibits near wild-type growth characteristics, infects cells via HER2/neu instead of CAR and represents an important step toward the development of genetically re-targeted adenoviruses with clinical relevance.

Efficient incorporation of a functional hyper-stable single-chain antibody fragment protein-IX fusion in the adenovirus capsid.

Recombinant adenoviruses are frequently used as gene transfer vehicles for therapeutic gene delivery. Strategies to amend their tropism include the incorporation of polypeptides with high affinity for cellular receptors. Single-chain antibodies have a great potential to achieve such cell type specificity. In this study, we evaluated the efficiency of incorporation of a single-chain antibody fused with the adenovirus minor capsid protein IX in the capsid of adenovirus type 5 vectors. To this end, the codons for the single-chain antibody fragments (scFv) 13R4 were fused with those encoding of pIX via a 75-Angstrom spacer sequence. The 13R4 is a hyper-stable single-chain antibody directed against beta-galactosidase, which was selected for its capacity to fold correctly in a reducing environment such as the cytoplasm. A lentiviral vector was used to stably express the pIX.flag.75.13R4.MYC.HIS fusion gene in 911 helper cells. Upon propagation of pIX-gene deleted human adenovirus-5 vectors on these cells, the pIX-fusion protein was efficiently incorporated in the capsid. Here, the 13R4 scFv was functional as was evident from its capacity to bind its ligand beta-galactosidase. These data demonstrate that the minor capsid protein IX can be used as an anchor for incorporation of single-chain antibodies in the capsids of adenovirus vectors.

Decreased immune reactivity towards a knobless, affibody-targeted adenovirus type 5 vector.

In this study, a prototype Adenovirus type 5 (Ad5) vector deleted of the fiber knob domain and carrying an Affibody molecule as the targeting ligand showed decreased susceptibility to human pre-existing antibodies. This vector, Ad5/R7-Z(taq)Z(taq), has short fibers carrying seven shaft repeats, a non-native trimerization signal and an affibody molecule (Z(taq)) reactive to Taq polymerase. Ad5/R7-Z(taq)Z(taq) could be specifically targeted to 293 cells stably expressing membrane-bound anti-Z(taq) idiotypic affibody called Z(ztaq) (293Z(ztaq)). Sera from 50 blood donors were analyzed for neutralization activity (NA) against the parental Ad5/Fiwt vector and knobless Ad5/R7-Z(taq)Z(taq) on 293Z(ztaq) cells. Twenty-three sera had NA titers (> or =1:64) against Ad5/Fiwt (46%) and only two against Ad5/R7-Z(taq)Z(taq) (4%). Characterization of sera with NA titers showed that the knob domain is one of the targets of the antibodies. Neutralization assays using sera pre-adsorbed on knob and hexon proteins showed that the NA of the sera was carried mainly by anti-knob and anti-hexon antibodies, but in certain sera the anti-hexon antibodies represent the major population of the neutralizing antibodies (NAbs). Our results suggested that a combination of knob deletion and hexon switching could be an effective strategy for Ad vectors to better evade the anti-Ad NAbs.

HIV/AIDS communication campaigns: progress and prospects.

The mass media have been the primary method for disseminating human immunodeficiency virus (HIV)/acquired immune deficiency syndrome (AIDS) prevention messages worldwide. In this article, we update previous reviews by systematically examining published articles (n = 41) of empirical evaluations of international HIV/AIDS prevention campaigns. Of the 41 studies identified, 17 countries are represented. In this review, we examine six components related to media campaign design and evaluation: target audience, communication channel(s), message content, campaign theme, exposure, and outcomes. Of the studies included in this sample, each one described the target audience; 93% reported on channel selection; 75% described message content; 63% mentioned a campaign theme; and 62% documented campaign exposure. We investigate also the extent to which HIV/AIDS prevention efforts have moved beyond media campaigns to comprehensive communitywide programs. We conclude that HIV/AIDS prevention efforts would benefit from: (1) better reporting of media campaign components and outcomes, (2) more systematic evaluation, (3) greater integration of theory, and (4) increased attention to communitywide intervention strategies.

Representation of orientation and spatial frequency in perception and memory: a choice reaction-time analysis.

Discrimination and short-term memory for the orientation of sinusoidal gratings that differed in spatial frequency, and for the spatial frequency of gratings that differed in orientation, were measured in a same-different task with 0-10 s interstimulus intervals (ISI) between test and reference stimuli. Introducing a difference between test and reference stimuli on a second dimension, or increasing ISI, did not impair spatial discrimination in terms of accuracy, but choice reaction times for correct decisions were prolonged by both manipulations. Results suggest that perceptual discrimination is based on representations in which orientation and spatial frequency are conjointly coded and that decisions are reached by a serial process scanning multiple-tuned, labeled channels; short-term memory may involve reactivation of these channels.

Cancer knowledge and perceptions among Chinese factory workers: implications for cancer control and prevention.

The global "war on cancer" increasingly depends on public perceptions of primary and secondary cancer prevention and control efforts. Recognition of cancer warning signs, understanding the importance of early diagnosis, and participation in cancer screening programs are partly functions of an individual's attitudes and knowledge. A questionnaire designed to determine present knowledge and attitudes towards a variety of cancer issues was administered to a random sample of Chinese factory workers in two port cities in southeast China. Our findings revealed that, like Americans, Chinese workers are uncertain and ill informed about many cancer issues. this study also demonstrates that cancer risk factors, cancer warning signs, and harmful occupational exposures are not common knowledge among the Chinese lay public. These findings should facilitate future chinese cancer education and control efforts and further the need among health education planners to recognize and appreciate public perceptions and misperceptions of a notably complex disease.

Developing health education materials for inner-city low literacy parents.

The question of identifying and treating childhood illness confronts all new parents. Misconceptions often lead parents to manage illnesses in their young children inappropriately through overly aggressive treatment or insufficient attention. This responsibility is especially challenging for low-income new parents who lack the literacy levels needed to understand and use much of the existing health education literature and who are without access to health facilities and providers. In response to a perceived need for health information directed at low-income, low-literacy parents, students from the University of California at Los Angeles School of Public Health created an easy-to-use reference booklet called "A Parent's Guide: When Your Child Is Sick." The booklet's aim is to assist parents in treating common childhood illness and identifying more serious diseases requiring medical attention. A comprehensive and manageable amount of information is provided in the booklet. Behaviors and issues covered include (a) recognition of symptoms, (b) actions that could be taken in the home, (c) medicines that could be administered, and (d) recommendations on how persistent problems might be handled.

New multiple congenital anomalies syndrome in a stillborn infant of consanguinous parents and a prediabetic pregnancy.

We report on a term stillborn female infant with multiple congenital anomalies (MCA) which have not previously been reported as occurring together. The malformations include a first and second branchial arch sequence, ectopia cordis with congenital heart defect, caudal "regression" sequence with absent sacrum and hypoplastic right femur, ectrodactyly, left radial abnormality, islet cell hyperplasia, and skin lesions. The pregnancy was complicated by abdominal cramping with exercise, heavy vaginal bleeding, maternal obesity, and a normal screening glucose tolerance test at 6 months gestation. The infant was born to 20-year-old G3P1SAB2 Mexican-American parents who are first cousins. There was strong maternal family history of adult-onset diabetes. The malformations have some findings in common with those seen in infants of diabetic mothers. Structural defects similar to, but not inclusive of, those in our infant have been reported in 2 sibs born to a prediabetic mother with a first cousin marriage as well as in focal dermal hypoplasia. Parental consanguinity is suggestive of an autosomal recessive disorder. Alternatively, it may represent a combined multifactorial effect making the conceptus more sensitive to metabolic teratogens and thus placing it at increased risk for disruption of normal development.

Amniotic fluid embolus. A report of two cases resulting in maternal survival.

Two cases of amniotic fluid embolus resulted in patient survival. Both were manifested by fetal bradycardia, respiratory distress and disseminated intravascular coagulopathy. Rapid diagnosis and treatment and anticipation of subsequent events, especially coagulopathy, appear to offer the best chance of a successful outcome.

Plant protoplasts immobilized in calcium alginate: a simple method of preparing fragile cells for transmission electron microscopy.

A simple method for electron microscopic preparation of plant protoplasts is described. The main problems in preparing these fragile protoplasts for electron microscopy have been cell collapse due to steep gradients between protoplasts and fixatives and unacceptable loss of material during the many steps of the procedure. These problems may be solved by immobilization of the protoplasts in calcium alginate beads. The free diffusion properties of this gel prevent steep gradients. The beads also simplify handling and prevent loss of material. Protoplasts isolated from hypocotyls of rape, Brassica napus (var. Niklas), have been used as a model system. Transmission electron microscopy of the immobilized protoplasts osmotically stabilized with glucose demonstrated adequate structural and ultrastructural preservation.

Ectopia cordis thoracalis with craniofacial defects resulting from early amnion rupture.

A 33-week-gestation fetus was evaluated as having ectopia cordis thoracalis, midline sternal cleft, frontonasal dysgenesis, a midfacial cleft, and amniotic bands. Fibrous bands were attached to the apex of the heart, face, and the brain. There were no associated limb defects or scoliosis. The findings in this fetus and those in the literature suggest that acute amnion rupture in the third week of gestation may be the cause of these defects. Mechanical teratogenesis by tissue bands adherent to the heart may lead to ectopia cordis. The changes seen in the thorax, face, and brain suggest pressure necrosis, incomplete morphogenesis, tearing and tethering by amnion, and compression as a cause for these defects. A spectrum of defects is found following amnion rupture with the abnormalities being dependent upon the timing of the event. Ectopia cordis with amniotic bands appears to have an etiology distinct from isolated ectopia cordis. This suggests several different etiologies for ectopia cordis.

Demonstration of extracellular chymotrypsin-like activity from various Legionella species.

Concentrated extracellular supernatants from Legionella pneumophila, L. bozemanii, L. dumoffi, L. gormanii, but not L. micdadei, exhibited a strong chymotrypsin-like activity upon synthetic chromogenic tri- and tetrapeptides. Bacterial cell sonic extracts showed low protease activities, different from those of the extracellular concentrates. Extracellular concentrates were also tested in the API ZYM system, where weak protease activity only was recorded. Protease inhibitors decreased the activity of the extracellular Legionella proteases upon the chromogenic peptides.