Characteristics of virus-specific CD8(+) T cells in the liver during the control and resolution phases of influenza pneumonia

Dissection of the primary and secondary response to an influenza A virus established that the liver contains a substantial population of CD8(+) T cells specific for the immunodominant epitope formed by H-2Db and the influenza virus nucleoprotein peptide fragment NP366-374 (DbNP366). The numbers of CD8(+) DbNP366(+) cells in the liver reflected the magnitude of the inflammatory process in the pneumonic lung, though replication of this influenza virus is limited to the respiratory tract. Analysis of surface phenotypes indicated that the liver CD8(+) DbNP366(+) cells tended to be more "activated" than the set recovered from lymphoid tissue but generally less so than those from the lung. The distinguishing characteristic of the lymphocytes from the liver was that the prevalence of the CD8(+) DbNP366(+) set was always much higher than the percentage of CD8(+) T cells that could be induced to synthesize interferon gamma after short-term, in vitro stimulation with the NP366-374 peptide, whereas these values were generally comparable for virus-specific CD8(+) T cells recovered from other tissue sites. Also, the numbers of apoptotic CD8(+) T cells were higher in the liver. The results overall are consistent with the idea that antigen-specific CD8(+) T cells are destroyed in the liver during the control and resolution phases of this viral infection, though this destruction is not necessarily an immediate process.     

Different patterns of truncated prion protein fragments correlate with distinct phenotypes in P102L Gerstmann-Str?ussler-Scheinker disease

The clinicopathological phenotype of the Gerstmann-Str?ussler-Scheinker disease (GSS) variant linked to the codon 102 mutation in the prion protein (PrP) gene (GSS P102L) shows a high heterogeneity. This variability also is observed in subjects with the same prion protein gene PRNP haplotype and is independent from the duration of the disease. Immunoblot analysis of brain homogenates from GSS P102L patients showed two major protease-resistant PrP fragments (PrP-res) with molecular masses of approximately 21 and 8 kDa, respectively. The 21-kDa fragment, similar to the PrP-res type 1 described in Creutzfeldt-Jakob disease, was found in five of the seven subjects and correlated with the presence of spongiform degeneration and "synaptic" pattern of PrP deposition whereas the 8-kDa fragment, similar to those described in other variants of GSS, was found in all subjects in brain regions showing PrP-positive multicentric amyloid deposits. These data further indicate that the neuropathology of prion diseases largely depends on the type of PrP-res fragment that forms in vivo. Because the formation of PrP-res fragments of 7-8 kDa with ragged N and C termini is not a feature of Creutzfeldt-Jakob disease or fatal familial insomnia but appears to be shared by most GSS subtypes, it may represent a molecular marker for this disorder.     

Entropy-based template analysis in face biometric identification systems

The accuracy of a biometric matching algorithm relies on its ability to better separate score distributions for genuine and impostor subjects. However, capture conditions (e.g. illumination or acquisition devices) as well as factors related to the subject at hand (e.g. pose or occlusions) may even take a generally accurate algorithm to provide incorrect answers. Techniques for face classification are still too sensitive to image distortion, and this limit hinders their use in large-scale commercial applications, which are typically run in uncontrolled settings. This paper will join the notion of quality with the further interesting concept of representativeness of a biometric sample, taking into account the case of more samples per subject. Though being of excellent quality, the gallery samples belonging to a certain subject might be very (too much) similar among them, so that even a moderately different sample of the same subject in input will cause an error. This seems to indicate that quality measures alone are not able to guarantee good performances. In practice, a subject gallery should include a sufficient amount of possible variations, in order to allow correct recognition in different situations. We call this gallery feature representativeness. A significant feature to consider together with quality is the sufficient representativeness of (each) subject’s gallery. A strategy to address this problem is to investigate the role of the entropy, which is computed over a set of samples of a same subject. The paper will present a number of applications of such a measure in handling the galleries of the different users who are registered in a system. The resulting criteria might also guide template updating, to assure gallery representativeness over time.     

A novel technique for management of the en bloc bladder cuff and distal ureter during laparoscopic nephroureterectomy

PURPOSE: The optimal technique of excising the juxtavesical ureter and bladder cuff during laparoscopic nephroureterectomy is still evolving. We report on a novel transvesical needlescopic (2 mm. instrumentation) assisted technique of en bloc retrieval of the juxtavesical ureter and bladder cuff during laparoscopic radical nephroureterectomy for upper tract transitional cell carcinoma. MATERIALS AND METHODS: Retroperitoneal laparoscopic nephroureterectomy was performed in 8 patients using this technique. Two needlescopic ports (2 mm.) inserted suprapubically into the bladder were used in combination with a cystoscopically positioned Collins knife. RESULTS: Satisfactory circumferential detachment of the bladder cuff and en bloc mobilization of 3 to 4 cm. of the intact pelvic extravesical ureter were achieved transvesically in each case in a manner comparable to open surgery. CONCLUSIONS: This technique simulates established open surgical principles of treating the distal ureter during laparoscopic nephroureterectomy.     

Serial sectioning of thick tissue with a novel vibrating blade microtome

Vibrating blade microtomes are used extensively in biological research to section non-frozen tissue. There are a wide variety of commercial instruments available for this purpose, however, they are designed to cut thin sections primarily from a tissue block less than one centimeter in height. Herein is described a simple modification of a microscope frame that creates a vibrating blade microtome capable of producing a sequential series of sections through three centimeters of tissue. We illustrate the use of this device to identify and reconstruct a column of rat spinal motor neurons retrogradely labeled from a peripheral muscle.     

β-Tricalcium phosphate 3D scaffold promote alone osteogenic differentiation of human adipose stem cells: in?vitro study

Human adipose tissues surgically resected from the subcutaneous abdominal region were enzymatically processed to obtain Human Adipose Stem cells (fibroblast-like adipose tissue-derived stromal cells—ADSC-FL) that were immunophenotypically characterized using a panel of mesenchymal markers by flow cytometry. The formation of new hydroxyapatite crystals in culture dishes, by differentiating cells, further demonstrate the osteogenic potential of purified cells. The aim of this study was to evaluate the osteogenic differentiation potential of ADSC-FL seeded onto a porous β-tricalcium phosphate (β-TCP) matrix. ADSC-FL was cultured on the β-TCP matrix in medium with or without osteogenic differentiation additives. Time-dependent cell differentiation was monitored using osteogenic markers such as alkaline phosphatase (activity assay), osteocalcin and ostopontin (ELISA method) expression. Our results reveal that β-TCP triggers the differentiation of ADSC-FL toward an osteoblastic phenotype irrespective of whether the cells are grown in a proliferative or a differentiative medium. Hence, a β-TCP matrix is sufficient to promote osteoblastic differentiation of ADSC-FL. However, in proliferative medium, alkaline phosphatase activity was detected at lower level respect to differentiative medium and osteocalcin and osteopontin showed an expression delay in cells cultured in proliferative medium respect to differentiative one. Moreover, we observed an increase in FAK phosphorylation at level of tyrosine residue in position 397 (Western-blot) that indicates a good cell adhesion to β-TCP scaffold. In conclusion, our paper demonstrates that a three-dimensional β-TCP scaffold in vitro triggers on its own the differentiation of ADSC-FL toward an osteoblastic phenotype without the need to use differentiative media.     

Today's minimal requirements for a practical dental office infection control and exposure control program

The current climate in society regarding infectious diseases in general, and herpes, hepatitis, and HIV infections in particular, dictates that today's dental practices must use effective infection control techniques. The Occupational Safety and Health Administration continues to inspect, cite, and fine health care facilities. More states are implementing regulations concerning the operation of health care facilities. Patients are becoming more sophisticated in their scrutinizing of the dental and medical professions' approach to asepsis. Media coverage of exposure incidents is becoming more intense. All these factors leave dentists no choice; they must implement appropriate infection control techniques. The life-time cost of effective infection control is far less than one malpractice settlement. Implementation of an effective infection control program to promote dental asepsis can be cost-effective. In addition, it can be a practice builder.     

Worm-diseases]

Intestinal parasites are common among labourers from foreign countries and in immigrants. Some worm-infections like ascaridiasis or enterobiasis occur in children quite often even in the cold climates. In cases of ascaridiasis only a heavy infection causes clinical symptoms. The diagnosis is made through the detection of eggs in the stools, as all the other worm-infections are diagnosed from the stool-test. Piperazine-derivatives are highly effective against ascaris; another useful drug is Pyrantel-Pamoate. Toxocara canis, which affects normally only dogs, can occur occasionally in humans. Its larvae migrate in the body and cause granulomas. The main clinical symptom of threadworms (enterobiasis) is itching around the anus. This parasite can be eradicated through Pyrvinium-Pamoate. The first symptom a patient notices in cases of taeniasis are proglottides--parts of the worms--which are passed with the stools. The drug of choice against all kinds of tapeworms is Niclosamide. Hookworms are rampant in tropical countries. Heavy infections can cause severe anemias, especially in children. Bephenium-Naphthoate and Bitoscanate are effective drugs against this parasite. The whipworm (Trichuris trichura) is very common in the warm countries. It can be treated by Tiabendazole and nowadays also with Mebendazole, which is virtually atoxic. Infections with Strongyloides stercoralis are difficult to eradicate, as there are different ways of reinfection. In case an infection with strongyloides is accompanied by other serious diseases cachexia and even fatal outcome has been seen.     

Urographic excretion studies with metrizamide and "Dimer": a high dose comparison in dogs

The effects of a training experience that involves foot shock on the endogenous phosphorylation of membrane-bound proteins from brain were studied. Crude membrane fractions were prepared from the cerebral cortex and neostriatum of animals that had been sacrificed by quick freezing. In vitro incubation of the membranes with gamma-32P-ATP, follwed by SDS-gel electrophoresis of the phosphorylated substrates, revealed that the phosphorylation of two protein components (designated F and H-I) increased in preparations from animals that were subjected to a training experience 24 hr prior to sacrifice. These effects were greater in preparations from the neostriatum than from the cerebral cortex, and were observed in experiments using both rats and mice. Although all trained animals showed a high phosphorylation of bands F and H-I, control animals showed a greater variability in the phosphorylation of these bands. The results indicate that the phosphorylation of specific proteins may play a mediatory role in the processing of experiential information.     

Blunting of the neurotensin mRNA response to haloperidol in the striatum of aging rats: possible relationship to decline in dopamine D2 receptor expression

Neuroleptic drugs such as haloperidol (H) induce a rapid increase in neurotensin/neuromedin N (NT/N) gene expression in the dorsolateral striatum (DLSt) and nucleus accumbens (NA) in young adult rats. This effect may be mediated by post-receptor effectors that are activated by dopamine D2 receptor antagonism. The regional pattern of induction of neurotensin gene expression correlates with the side effect profile of particular neuroleptics. As motor side effects of H differ in aged animals, we hypothesized that the regional expression of the neurotensin gene may differ between young and old animals. We administered H or saline acutely to 3, 14, and 25 month-old Fischer 344 rats, followed by in situ hybridization and quantitative autoradiography for NT/N mRNA. There was a significant age effect on the H-induced NT/N mRNA response in the DLSt, but not the NA, of older animals. In addition to the blunted NT/N mRNA response, significant decreases in D2 receptor mRNA were observed in the lateral striatum of another group of young, middle-aged, and aged rats. Age-related blunting of the NT/N mRNA response to H in the DLSt may be due in part to a decrease in D2 receptors in this structure.