IFNτ mediates chemotaxis, motility, metabolism and CK18 downregulation in bovine trophoblast cells in vitro via STAT1 and MAPK42/44 signaling.

INTRODUCTION: IFNτ is the ruminant pregnancy recognition signal. In the study we investigated the autocrine influence of IFNτ on bovine F3 trophoblast cells. In detail chemotaxis, motility, metabolism, cell polarisation (CK18; ezrin) and the underlying classical (STAT1) and non-classical (MAPK42/44) signaling pathways were examined. METHODS: Cellular signaling was analysed by densitometric Western blot (STAT1, MAPK42/44, proteinkinase B) and RT-PCR (IFNAR1, -2). Cellular assays were carried out for chemotaxis (agarose spot assay), cell motility (live cell imaging), metabolism (MTT) and cell polarisation (CK18; ezrin). In vivo-produced conceptuses of gestational days (GD) 20-39 underwent immunohistochemistry (CK18; ezrin) to set the in vitro findings (cell polarisation) in proportion to the in vivo situation. RESULTS: IFNτ (10-1000 ng/ml) mediated dose-dependent effects. 10 ng/ml IFNτ induced chemotaxis and motility, whilst 1000 ng/ml led to reduced chemotaxis, motility and a 92-fold activation of MAPK44. Stimulation of cells with 10-1000 ng/ml IFNτ promoted metabolism (1.4-fold), increased the gene expression of IFNAR1/2 (24 h) and downregulated CK18 but not ezrin. All described in vitro effects were significant. Signaling, motility and metabolism could be blocked by specific inhibitors (PD98059, LY294002). CK18 and ezrin expression patterns in the trophoblast of in vivo conceptuses differed depending on GD. DISCUSSION: IFNτ is a major factor for bovine F3 trophoblast cells and mediates a variety of cellular actions ranging from chemotaxis to polarisation. IFNτ exerts its effects via classical (STAT1) and non-classical (MAPK42/44) signaling pathways in a dose-dependent way. We hypothesize that (dose-dependent) IFNτ regulation of the cellular effects could also be essential for bovine elongation and implantation.

Challenges in clinical teaching investigated: Towards sustainable staff recruitment and retention in dentistry.

PURPOSE: The study was conducted to identify possible factors and potential barriers that affect the retention and recruitment of clinical teaching staff at the Faculty of Dentistry, University of Otago, New Zealand. METHODOLOGY: A short questionnaire survey was distributed to 47 clinical teaching staff to understand the retention strategies and barriers. The collected quantitative and qualitative data were analysed and presented. FINDINGS: The response rate was 96%, with an equal distribution of female and male clinicians. The majority of participants were European New Zealanders. Themes influencing retention strategies were the necessity of formal teaching skills, clinical teaching as a career pathway, pay progression, lack of support and heavy workload. PRACTICAL IMPLICATIONS: Results suggested that generally the respondents to this survey were motivated to give back to their profession through engaging in dental clinical teaching. This study identified that lack of a career development and minimal pay progression are major factors impacting on the retention of clinical teaching staff.

Estimation of jaw-opening forces in adults.

OBJECTIVES: To estimate maximum jaw-opening forces in healthy participants of diverse ancestry and to estimate whether opening forces are associated with sex, age and anthropometric parameters such as height, weight and BMI. SETTING AND SAMPLE POPULATION: One hundred and forty-nine participants aged 20-60 years with overall good oral and general health. Exclusion criteria included myofascial or neck pain, symptomatic temporomandibular joint disorders (TMD), current orthodontic treatment or the absence of a natural dentition. MATERIAL AND METHODS: Jaw-opening forces were measured with an adjustable rigid extra-oral device connected to a 1000 N load cell. Seven attempts were recorded, with 10 seconds interval. Median force values were obtained after discarding the first and last attempt. The height and weight of each participant were measured and recorded, alongside age, sex and ethnicity. RESULTS: Men had greater maximum opening force median values than women (P < .001). Median (IQR) values for women were 41.16 N (30.44) and 79.00 N for men (63.86). Jaw-opening force values were poorly associated with biological and anthropometric parameters. CONCLUSION: In this study, which included a large sample of participants of broad age range and from a demographically diverse background, jaw-opening force values were greater in males than in females; however, force values were poorly associated with biological and anthropometric parameters. Future studies should explore the potential of this method as a screening tool for TMJ disorders and other conditions.

Exchanging the active site between phytases for altering the functional properties of the enzyme.

By using a novel consensus approach, we have previously managed to generate a fully synthetic phytase, consensus phytase-1, that was 15-26 degrees C more thermostable than the parent fungal phytases used in its design (Lehmann et al., 2000). We now sought to use the backbone of consensus phytase-1 and to modify its catalytic properties. This was done by replacing a considerable part of the active site (i.e., all the divergent residues) with the corresponding residues of Aspergillus niger NRRL 3135 phytase, which displays pronounced differences in specific activity, substrate specificity, and pH-activity profile. For the new protein termed consensus phytase-7, a major - although not complete - shift in catalytic properties was observed, demonstrating that rational transfer of favorable catalytic properties from one phytase to another is possible by using this approach. Although the exchange of the active site was associated with a 7.6 degrees C decrease in unfolding temperature (Tm) as measured by differential scanning calorimetry, consensus phytase-7 still was >7 degrees C more thermostable than all wild-type ascomycete phytases known to date. Thus, combination of the consensus approach with the selection of a "preferred" active site allows the design of a thermostabilized variant of an enzyme family of interest that (most closely) matches the most favorable catalytic properties found among its family members.

Structure, properties, and tissue localization of apoplastic alpha-glucosidase in crucifers.

Apoplastic alpha-glucosidases occur widely in plants but their function is unknown because appropriate substrates in the apoplast have not been identified. Arabidopsis contains at least three alpha-glucosidase genes; Aglu-1 and Aglu-3 are sequenced and Aglu-2 is known from six expressed sequence tags. Antibodies raised to a portion of Aglu-1 expressed in Escherichia coli recognize two proteins of 96 and 81 kD, respectively, in vegetative tissues of Arabidopsis, broccoli (Brassica oleracea L.), and mustard (Brassica napus L.). The acidic alpha-glucosidase activity from broccoli flower buds was purified using concanavalin A and ion-exchange chromatography. Two active fractions were resolved and both contained a 96-kD immunoreactive polypeptide. The N-terminal sequence from the 96-kD broccoli alpha-glucosidase indicated that it corresponds to the Arabidopsis Aglu-2 gene and that approximately 15 kD of the predicted N terminus was cleaved. The 81-kD protein was more abundant than the 96-kD protein, but it was not active with 4-methylumbelliferyl-alpha-D-glucopyranoside as the substrate and it did not bind to concanavalin A. In situ activity staining using 5-bromo-4-chloro-3-indolyl-alpha-D-glucopyranoside revealed that the acidic alpha-glucosidase activity is predominantly located in the outer cortex of broccoli stems and in vascular tissue, especially in leaf traces.

Distribution of nitric oxide synthase implies a regulation of circulation, smooth muscle tone, and secretory function in the human prostate by nitric oxide.

BACKGROUND: Nitric oxide (NO) is suggested as a mediator involved in the regulation of smooth muscle tone, blood flow, and secretory function of the genitourinary tract and originates from different NO synthase (NOS) isoforms located in endothelial, neuronal, and epithelial structures. The aim of the present study was to determine the location of endothelial and neuronal NOS in the human prostate. METHODS: Histochemical NADPH-diaphorase (NADPH-d) staining, ultrastructural NADPH examination, and NOS immunohistochemistry were performed on histologically verified nonmalignant prostate tissue from normal nonobstructive and hyperplastic obstructive human prostates. RESULTS: In the prostatic tissue, NADPH-d staining and immunohistochemistry with bNOS antibody revealed the existence of a dense nitrinergic innervation of glandular epithelium, fibromuscular stroma, and blood vessels. NADPH-d reaction in glandular epithelium was not confirmed by ecNOS or bNOS immunohistochemistry. In benign prostatic hyperplasia (BPH), the nitrinergic innervation is reduced. The vascular distribution of ecNOS provides evidence for a segmental differentiation of the NO-mediated vascular regulation. CONCLUSIONS: NO plays an important role in the autonomic innervation of all compartments of prostatic tissue. In obstructive BPH, the nitrinergic innervation is reduced compared to that in normal prostate tissue. Further studies are necessary to elucidate the complex role of NO in the prostate.

[Pattern of distribution of constitutive isoforms of NO synthase in the normal prostate and obstructive prostatic hyperplasia]. / Verteilungsmuster der konstitutiven Isoformen der NO-Synthase in der normalen Prostata und der obstruktiven Prostatahyperplasie.

Nitric oxide (NO) is suggested as an important mediator for the regulation of biological processes. In the present study we tried to determine histochemically and immunohistochemically the localization and distribution of the constitutive NO-synthase Isoforms (bNOS and eNOS) of 14 normal non-obstructive and 12 hyperplastic obstructive human prostates. Differentiated nitrinergic innervation was shown for the prostate glands, fibromuscular stroma and blood vessels by NADPH-diaphorase staining and immunohistochemically with specific NOS antibodies. In the specimens with benign prostatic hyperplasia nitrinergic innervation seems to be distinctly reduced. The vascular distribution of NOS provides evidence for segmental differentiation of NO-mediated vascular regulation. The NADPH-diaphorase reaction was not confirmed immunohistochemically by the specific NOS antibody in the glandular epithelium. The distribution of NO synthase shows the importance of nitric oxide in the regulation of smooth muscle tone, blood flow and secretory function in the normal and hyperplastic human prostate.

[Studies on the compression of the external carotid artery in the region of the styloid process of the temporal bone]. / Untersuchungen zur Kompression der A. carotis externa im Bereich des Processus styloideus ossis temporalis.

The styloid process syndrome and the stylo-keratohyoid syndrome are well-known diseases caused by an elongated styloid process of the temporal bone or by ossification of the derivations of the second branchial arch. The cause of the complaint is a compression of neighbouring vessels, nerves and soft tissues. 85 anatomical specimens were investigated in respect of a compression of the external carotid artery by the stylohyoid muscle. In 7 cases we found an impression of the vessel caused solely by the stylohyoid muscle without ossification of the derivations of the second branchial arch or elongation of the styloid process. In some cases the posterior auricular artery was also compressed. In addition, the extent of the stenosis of the external carotid artery was examined quantitatively. At least a histological examination was performed. The symptoms of the diseases are discussed by a review of the literature. Difficulties in X-ray diagnosis or diagnosis by bimanual palpation are pointed out.

Compensation for beam intensity fluctuation in determination of P(ion) the ion-recombination correction factor for ionization chambers, by the two-voltage technique.

We have developed a method of compensation for fluctuations in beam intensity that may occur during measurement of Pion, the ion-recombination correction factor, by the two-voltage technique. The method requires signals proportional to beam intensity during measurement. We used a parallel-plate ionization chamber, whose Pion was known, and a vacuum chamber to obtain signals that were proportional to the beam intensity. Experiments were conducted using pulsed proton beam providing doses that ranged from 0.16 to 0.01 cGy/pulse. The value of Pion of a thimble ionization chamber was measured. With these measurements, the validity of the method which we proposed for pulsed beam was verified experimentally.

Ultrastructural studies of the hemocytes of Panstrongylus megistus (Hemiptera: Reduviidae).

Ultrastructural analyses revealed the presence of six hemocyte types in the hemolymph of Panstrongylus megistus, partially confirming our previous results obtained through light microscopy. Prohemocytes: small, round hemocytes with a thin cytoplasm layer, especially rich in free ribosomes and poor in membranous systems. Plasmatocytes: polymorphic cells, whose cytoplasm contains many lysosomes and a well developed rough endoplasmic reticulum (RER). They are extremely phagocytic. Sometimes, they show a large vacuolation. Granulocytes: granular hemocytes whose granules show different degrees of electrodensity. Most of them, have an internal structuration. Coagulocytes: oval or elongated hemocytes, which show pronounced perinuclear cisternae as normally observed in coagulocytes. The cytoplasm is usually electrodense, poor in membranous systems and contains many labile granules. Oenocytoids: large and very stable hemocytes, whose homogeneous cytoplasm is rich in loose ribosomes and poor in membranous systems. Adipohemocytes: large cells, containing several characteristic lipid droplets. The cytoplasm is also rich in glycogen, RER and large mitochondria. The total and differential hemocyte count (THC and DHC) were also calculated for this reduviid. THC increases from 2,900 hemocytes/mm3 of hemolymph in the 4th instar to 4,350 in the 5th and then, decreases to 1,950 in the adults. Plasmatocytes and coagulocytes are the predominant hemocyte types.

Utilidad del tubo de drenaje en la peritonitis difusa / Use of tube drains in diffuse peritonitis

Se presenta un estudio prospectivo y randomizado sobre 40 pacientes con peritonitis difusa, intervenidos en el SUAO. A 20 de ellos se les dejó tubo de drenaje como parte del tratamiento quirúrgico. Los grupos en estudio son comparativamente similares. El uso del tubo de drenaje significó un retardo en la recuperación del tránsito intestinal, una evolución febril mas prolongada y patología agregada. La morbilidad intraabdominal no fue prevenida con el uso del tubo de drenaje y significó un 20% mayor, derivada de éste. Si la técnica quirúrgica trata el foco y se efectúa un prolijo aseo de la cavidad abdominal, creemos que en las peritonitis difusas es innecesario dejar tubo de drenaje y además produce complicaciones, razones por las que en estas circunstancias no lo usamos

The physical performances of a single slice positron tomographic system and preliminary results in a clinical environment.

Metabolic phenomena can be studied and measured non-invasively using positron emitting radionuclides and a suitably adapted tomographic system. The choice of a single slice ring camera is justified by its physical performance, which is presented here and discussed. A series of measurements with geometrical phantoms and analytical simulations have been performed to determine the critical characteristics of the system. This has permitted optimization of certain parameters enabling very interesting clinical results to be obtained at SHFJ, particularly in the area of cerebral physiopathology. In addition, the potential of obtaining absolute quantitative values of regional activity is presented. The calibration of the regional activity is presented. The calibration of the system, spatial non-stationarities, and attenuation correction, which represent the main sources of error, are considered in detail. A precision of the order of 10% should be obtainable. Such a quantitation method has been successfully applied to the in vivo study of the regional extraction of cerebral oxygen.