Safety evaluation of a novel muramidase for feed application.

Safety evaluation of a muramidase produced by a Trichoderma reesei strain (safe lineage), expressing a muramidase gene isolated from Acremonium alcalophilum is presented. Intended use in feed of this enzyme is as digestive aid in broiler chickens. Muramidase 007, was non-mutagenic and non-clastogenic in vitro, and no adverse effects were observed in 90-day subchronic toxicity studies in rats at doses up to 1132 mg TOS/kg body weight/day. The enzyme did not exhibit, in vitro, skin, nor eye irritation potential. Acute aquatic toxicity evaluated on daphnia and algae showed absence of effect of the enzyme at the standard doses tested. Muramidase 007 was fully tolerated by broiler chickens in a 6-weeks tolerance study showing no adverse effects in any of the dietary treatments (0, 1×, 5× and 10× maximum recommended dose). In conclusion, Muramidase 007 is found to be toxicologically inert, and there are no worker's safety concerns if standard precautions are instituted and a non-dusty formulation is employed. Muramidase 007 is well tolerated by the target species (broiler chickens) and cause no harm to the environment. The beneficial safety evaluation of Muramidase 007 is in line with this type of enzyme that is found ubiquitously in nature.

Toxicological studies on a novel phytase expressed from synthetic genes in Aspergillus oryzae.

Phytases are widely used as feed additives for monogastric animals, which cannot easily utilise the phosphorus bound in phytate (myo-inositol hexakisphosphate). The current study presents a safety evaluation of a 6-phytase produced by an Aspergillus oryzae strain expressing two synthetic genes, both mimicking a phytase gene from a Citrobacter braakii strain. Oral administration of the phytase preparation to rats at a dose level of 0.86 g total organic solids/kg body weight/day for 13 weeks did not cause any adverse effect. The phytase preparation did not exhibit irritative potential when applied locally to the eyes of rabbits or when applied to the skin using the in vitro three-dimensional epidermis model of adult human-derived epidermal keratinocytes. Furthermore, the phytase preparation was found not to represent mutagenic or clastogenic potential in the bacterial reverse mutation assay and in the in vitro micronucleus assays. Based on the toxicological data, the large safety factors calculated under common recommended dose assumptions for broiler chickens and weaned piglets, and the fact that Aspergillus oryzae is considered a safe strain lineage, it is concluded that there are no reasons for safety concerns when using this phytase as a feed additive.

Spontaneous fusion between cancer cells and endothelial cells.

Endothelial cells line the inside of blood and lymphatic vessels, and cancer cells must cross this barrier, first to gain access to the circulation, and, second, to exit and metastasize. How this occurs is incompletely understood. We now demonstrate that human cancer cells are able to fuse with endothelial cells to form hybrid cells displaying proteins and chromosomal markers characteristic of both parent cells. The hybrid cells are viable and capable of undergoing mitosis. Fusions between cancer cells and endothelial cells were shown to occur both in vitro, in co-cultures of human breast cancer cells and endothelial cells, and in vivo, following intravascular dissemination of human breast cancer cells in nude mice. These observations demonstrate a new type of cancer-endothelial cell interaction that may be of fundamental importance to the process of metastasis.

Multi-layer microfluidic glass chips for microanalytical applications.

A new, versatile architecture is presented for microfluidic devices made entirely from glass, for use with reagents which would prove highly corrosive for silicon. Chips consist of three layers of glass wafers bonded together by fusion bonding. On the inside wafer faces a network of microfluidic channels is created by photolithography and wet chemical etching. Low dead-volume fluidic connections between the layers are fabricated by spark-assisted etching (SAE), a computer numerical controlled (CNC)-like machining technique new to microfluidic system fabrication. This method is also used to form a vertical, long path-length, optical cuvette through the middle wafer for optical absorbance detection of low-concentration compounds. Advantages of this technique compared with other, more standard, methods are discussed. When the new glass-based device for flow-injection analysis of ammonia was compared with our first-generation chips based on silicon micromachining, concentration sensitivity was higher, because of the longer path-length of the optical cuvette. The dependence of dispersion on velocity profile and on channel cross-sectional geometry is discussed. The rapid implementation of the devices for an organic synthesis reaction, the Wittig reaction, is also briefly described.

Demographic variables and microCog performance: relationships for VA patients under treatment for substance abuse.

The associations of demographic variables with cognitive performance, as measured by the MicroCog ability domain scores, were assessed with 222 substance abusers who were patients in a VA medical center. Analysis indicated that age was negatively related to all five outcomes scores, and education was positively related to measures assessing Attention/Mental Control and Reasoning/Calculation. These findings are consistent with past theoretical and applied research. The demographic variables, as a group, accounted for a total of 17% to 37% of the variance in the five domain scores.

Sample preconcentration by field amplification stacking for microchip-based capillary electrophoresis.

A microchip structure for field amplification stacking (FAS) was developed, which allowed the formation of comparatively long, volumetrically defined sample plugs with a minimal electrophoretic bias. Up to 20-fold signal gains were achieved by injection and separation of 400 microm long plugs in a 7.5 cm long channel. We studied fluidic effects arising when solutions with mismatched ionic strengths are electrokinetically handled on microchips. In particular, the generation of pressure-driven Poiseuille flow effects in the capillary system due to different electroosmotic flow velocities in adjacent solution zones could clearly be observed by video imaging. The formation of a sample plug, stacking of the analyte and subsequent release into the separation column showed that careful control of electric fields in the side channels of the injection element is essential. To further improve the signal gain, a new chip layout was developed for full-column stacking with subsequent sample matrix removal by polarity switching. The design features a coupled-column structure with separate stacking and capillary electrophoresis (CE) channels, showing signal enhancements of up to 65-fold for a 69 mm long stacking channel.

Listening, understanding and interpreting: reflections on complexity.

Beginning with the premise that a contemporary approach to analytic exchanges has become far more complex and multi-faceted than in earlier times, the author addresses listening, understanding and interpreting. The opening section presents the basis and utility of a proposed theory of five motivational systems and identifies the need for a shift from conceptualising 'structures' to systems. The nature of communication during analysis is considered from several standpoints including listening for needs and intentions, the place of theory as a background to listening, an optimal state for analysand and analyst, and the significance of the distinction between inner monologue and spoken discourse. Differing views of free association and narrative, especially questions arising from findings of the 'adult attachment interview', are discussed. The patient's sensitivity to the presence and influence of the analyst and the analyst's recognition of non-verbal as well as verbal communications completes this section. In the final section a brief clinical example is presented to introduce a differentiated depiction of the variety of interventions that analysts employ. Throughout the paper, the author presents his view of listening, understanding and interpreting in dialectic contrast with the many other perspectives held by analysts in this period of theoretical pluralism.

The rat Subcutaneous Air Sac model: a quantitative assay of antiangiogenesis in induced vessels.

A new in vivo experimental model--the Subcutaneous Air Sac (SAS) model-has recently been presented to replace a previous in vivo rabbit cornea assay where neovascularisation was induced by chemical injury of the cornea or by implantation of tumour cells intracorneally, a methodology which is believed to cause severe pain to the animals. In the SAS model, an air sac is induced by injection of air subcutaneously on the back of the animal. After 10-14 days the air sac appears as an almost transparent avascular membrane in which induction of new vessels can be studied. We present recent developments of this technique: In the SAS-tumour technique, vascular endothelial growth factor-producing tumour cells are inoculated subcutaneously directly on the membrane, and the formation of new vessels is measured 8 days later. In the SAS-pellet technique, slow-release pellets containing angiogenic factors, basic fibroblast growth factor or vascular endothelial growth factor are implanted on the subcutaneous membrane by a simple operation. The formation of new vessels is measured 10 days later. The ability of the SAS-tumour- and SAS-pellet techniques to detect an antiangiogenic effect of a systemically administered compound was investigated using the fumagillin analogue TNP-470 (o-chloroacetyl-carbamoyl)-fumagillol) as a positive control given subcutaneously for 7 and 9 days, respectively. At a dose of 10 mg TNP-470/kg/day the angiogenesis was reduced by approximately 70% in the SAS-tumour technique and by 40-60% in the SAS-pellet technique. The animals were unaffected by the SAS methodology. The SAS-tumour and SAS-pellet models are considered complementary and make use of simple and almost similar techniques which facilitate the evaluation.

General principles of self psychology: a position statement.

In the more than twenty-five years since Kohut formulated a psychology of the self, the basic theory has undergone many revisions and additions. In the course of broadening from a focus on narcissism and empathy into a general theory of normal and pathological development, self psychology has taken so many different directions that the question can be asked, Does self psychology remain essentially a single theory with different descriptors--a theory of a self-selfobject matrix, a theory of intersubjectivity, a theory of motivational systems, and so on? A concise statement of general principles is intended to contribute to a dialogue between advocates of the views presented here and those who hold different views of theory and practice, within and withoutself psychology.

The rat subcutaneous air sac model: a new and simple method for in vivo screening of antiangiogenesis.

An experimental rat model, the Subcutaneous Air Sac (SAS) model, was developed to provide an animal model in which neo-vascularization can be easily assessed in situ and quantified using a radiolabelled plasma marker. The SAS model was designed to replace a previous model where neovascularization was induced by chemical injury of rat or rabbit cornea or by implantation of tumour cells intracorneally, a methodology which is believed to cause severe pain to the animals. In the SAS model the air sac replaces the cornea as a transparent avascular substratum in which vascularization can be observed. The air sac is induced by injection of air subcutaneously on the back of the animal. After 8 to 10 days a sufficient air sac has been established. The animal is anaesthesized and by a minor operation the cellulose sponge is implanted upon the air sac under the skin. The vasoproliferative effect of the cellulose sponge causes formation of new vessels which are macroscopically visible 10 days after implantation. The ability of the in vivo SAS model to show an antiangiogenic effect of a systemically applied test compound was investigated using the fumagillin analogue TNP-470 (ochloro-acetylcarbamoyl)-fumagillol) as a positive control at dose levels of 0, 1, 2.5, 5 and 10 mg/kg/day given subcutaneously for 10 days. The neo-angiogenesis was scored both in situ using a subjective point system and by measuring the 125I-activity of the implant and the membrane after an intravenous injection of 125I-labelled antibodies. The neo-angiogenesis was reduced by approximately 45-50% in animals treated with 5 or 10 mg/kg/day of TNP-470 compared to animals treated with the vehicle. The animals treated with 10 mg/kg/day TNP-470 showed signs of toxicity. The SAS model is considered highly relevant for in vivo testing of potential antiangiogenic drugs on humane grounds. The high reproducibility, the low cost and the technical simplicity of the method makes it attractive.

Toxicity of 1,25-dihydroxyvitamin D3, tacalcitol, and calcipotriol after topical treatment in rats.

To investigate and compare the effects on calcium metabolism of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) and the synthetic vitamin D analogs tacalcitol and calcipotriol after topical treatment, we treated groups of rats topically once daily with three dose levels of each test compound in a vehicle of propylene glycol plus ethanol for 28 d. The urinary calcium excretion was measured after 14 d, and serum calcium and parathyroid hormone were measured at termination. The rats were autopsied, and the kidneys were examined microscopically for mineralizations. Based on the urinary calcium excretion and the serum calcium level, calcipotriol was found to be 60 times less calcemic, and tacalcitol was slightly less calcemic, than 1,25(OH)2D3 after repeated topical application to rats. Serum parathyroid hormone was suppressed to a lower degree by calcipotriol and tacalcitol than by 1,25(OH)2D3, and the incidence and severity of renal corticomedullary mineralization were higher in rats treated with 1,25(OH)2D3 and tacalcitol than with calcipotriol. We conclude that calcipotriol is much less calcemic than 1,25(OH)2D3 or tacalcitol when applied topically to rats in a vehicle that enhances penetration into the skin. We attribute the lower calcemic effect of calcipotriol to the pharmacokinetic profile of the compound, particularly its rapid metabolization into inactive compounds.

When educators confront child abuse: an analysis of the decision to report.

Five scenarios of child abuse were used to study the recognition and reporting of child abuse in a sample of 664 teachers, counselors, school psychologists, principals, and district superintendents. The following results emerged: (a) Reporting tendency varied by type of abuse described, forming a 3-level hierarchy; (b) reporting tendency and reporting rate were unrelated to the gender of the victim or respondent; (c) reporting tendency was unrelated to the profession of the educator (i.e., principal, counselor, etc.), though certain types of abuse were suspected and/or reported significantly less often by classroom teachers; (d) for each scenario a linear composite of decisional items discriminated Reporters from Nonreporters with 75% to 84% accuracy. Most salient in distinguishing Reporters from Nonreporters were issues involving quality of suspicion and the respondents belief that schools should be a first line of defense against abuse and neglect; (e) educators were uniform in their high level of awareness of mandatory reporting laws; (f) educators preparedness to detect child abuse differed by profession, but most desired additional training. The implications of these findings are reviewed and suggestions made for revisions to social service policies and training for educators.

[Survival in patients after surgical closure of post-infarction rupture of the ventricular septum]. / Prezívání nemocných po chirurgickém uzáveru poinfarktové ruptury mezikomorové prepázky.

The authors discuss in the introduction hitherto reported data on the incidence and course of post-infarction ruptures of the interventricular septum and the results of conservative treatment. The high mortality rate after the conservative procedure led to attempts to resolve post-infarction ruptures of the interventricular septum surgically. The authors discuss in detail the indications for early and delayed operations has a decisive influence on the patients' survival. The authors had the opportunity to operate since 1978 till February 1986 five patients with post-infarction perforation of the interventricular septum. Four of the patients were women, one was a man. The mean age of the operated patients was 61 years, the oldest patient was a 75-year-old man and the youngest a 46-year-old woman. During the last check-up in 1994 it was revealed that of five operated patients three had died. One of them three years after operation and two after seven years. One female patient survives for 13 years and one for 11 years. Both are in a satisfactory conditions.

[Aneurysms of the left atrium and left atrial appendage]. / Aneuryzmata levé srdecní síne a levého ouska.

BACKGROUND: The authors deal with the problem of rare congenital and acquired aneurysm of the left atrium and auricular appendage. The authors attempted to compare findings on operation with results of preoperative examinations and evaluate the yield of echocardiographic and angiocardiographic examinations. METHODS AND RESULTS: The authors had the opportunity to encounter this unusual finding of cardiac aneurysms in five patients operated in 1987-1990 on account of an advanced mitral defect associated in two patients with ischaemic heart disease. In all patients acquired aneurysms were involved. In two a large thrombus was found. CONCLUSIONS: In the diagnosis of aneurysms of the left atrium and auricular appendage transoesophageal echocardiography proved to be the most reliable method.

[Advantages of lung surgery via medial sternotomy and use of extracorporeal circulation in very advanced lung tumors]. / Výhody plicních operací provádených z mediální sternotomie a uplatnení mimotelního obehu u znacne pokrocilého nádoru plic.

The authors present their initial experience with surgery of pulmonary affections in 32 patients where they used medial sternotomy as the route of approach. They provide evidence of the advantages of this approach, in particular in high-risk patients, and of the favourable effect of this approach on the postoperative course. Successful radical surgery under conditions of extracorporeal circulation, although a very advanced tumour was involved where posterolateral thoracotomy was unequivocally contraindicated, indicates that there is a possibility to extend indications of surgical therapy of otherwise inoperable pulmonary tumours.

A motivational systems approach to the clinical experience.

A conception of the self and five motivational systems is applied to clinical psychoanalysis. Each motivational system develops in infancy from innate and learned patterns in response to a basic need, and each involves particular affects. Each motivational system contributes patterns from which important transferences evolve. At any given moment, motives derived from one or another system dominate a person's experience, motives from the other system being subsidiary or dormant. We describe the manner in which these concepts contribute to an explanation of foreground-background relations during analysis, and how analysts and analysands construct model scenes to give meaning to information acquired by empathic listening. We conclude with a clinical vignette illustrating the application of these concepts to the patient's transference and the analyst's response in the intersubjective realm of an analytic enactment and verbal exchange.