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[Preliminary data on the behavior of the enzyme of metabolism of phospholipid bases in human tumor preparations].

The A.A. have investigated the phospholipids base-exchange enzyme system in the solid tumours in order to state if a correlation between this activity and the variation of the cellular ciclic nucleotides amount was possible considering that these compounds have been reported to undergo a variation in tumour compared with normal tissues. They report some previous results in a lung tumour and in an endometrial carcinoma, were they have found a big increase in the PhS synthesis. Such increase was possible to be seen in the endometrial carcinoma only after a stimulation by 17-beta-estradiol and it was reversed in this case by doxorubicine. These results suggest that an alteration of the PhS synthesis should be one of numerous peculiarity of the neoplastic cell.

Adenocarcinoma↗

New data on Myxobolus longisporus (Myxozoa: Myxobolidae), a gill infecting parasite of carp, Cyprinus carpio haematopterus, from Chinese lakes.

The original description of Myxobolus longisporus Nie et Li, 1992, the species infecting gills of Cyprinus carpio haematopterus L., is supplemented with new data on the spore morphology and pathogenicity. Spores are elongate pyriform with pointed anterior end, 15.7 (15.5-16.5) microm long, 6.7 (6-8) microm wide and 5.5 microm thick. Sutural ridge is straight and narrow. Mucus envelope is lacking. Two equal-sized elongate pyriform polar capsules are 8.5 microm long and 2.5 microm wide with convergent long axes. Polar filament coiled perpendicularly to the long axis of the capsule makes 9 (8-10) turns. Posterior end of polar capsules exceeds mid-spore by 15-20%. Cyst-like plasmodia are localised in the gill secondary lamellae. The infection is described in adult big host specimens. Gross lesions manifested as dark red colouration of gill tissues were restricted to the ventral part of the first gill arches. Remarkable site specificity (apical part of secondary lamellae) was observed in the course of development of microscopic lesions. M. longisporus is characterised also on the molecular level using sequences of SSU rRNA gene. Phylogenetic analysis based on these sequences has allowed clearer phylogenetic relationships to be established with other species of the genus Myxobolus sequenced to date.

Animals↗

A novel soluble protein factor with non-opioid dynorphin A-binding activity.

A novel soluble non-opioid dynorphin A-binding factor (DABF) was identified and characterized in neuronal cell lines, rat spinal cord, and brain. DABF binds dynorphin A(1-17), dynorphin A(2-17), and the 32 amino acid prodynorphin fragment big dynorphin consisting of dynorphin A and B, but not other opioid and non-opioid peptides, opiates, and benzomorphans. The IC50 for dynorphin A(1-17), dynorphin A(2-17), and big dynorphin is in the 5-10 nM range. Using dynorphin A and big dynorphin fragments a binding epitope was mapped to dynorphin A(6-13). DABF has a molecular mass of about 70 kDa. SH-groups are apparently involved in the binding of dynorphin A since p-hydroxy-mercuribenzoic acid inhibited this process. Upon interaction with DABF dynorphin A was converted into Leu-enkephalin, which remained bound to the protein. These data suggest that DABF functions as an oligopeptidase that forms stable and specific complexes with dynorphin A. The presence of DABF in brain structures and other tissues with low level of prodynorphin expression suggests that DABF as an oligopeptidase may degrade other peptides. Dynorphin A at the sites of its release in the CNS may attenuate this degradation as a competitor when it specifically binds to the enzyme.

Animals↗

Shrinkage estimators for covariance matrices.

Estimation of covariance matrices in small samples has been studied by many authors. Standard estimators, like the unstructured maximum likelihood estimator (ML) or restricted maximum likelihood (REML) estimator, can be very unstable with the smallest estimated eigenvalues being too small and the largest too big. A standard approach to more stably estimating the matrix in small samples is to compute the ML or REML estimator under some simple structure that involves estimation of fewer parameters, such as compound symmetry or independence. However, these estimators will not be consistent unless the hypothesized structure is correct. If interest focuses on estimation of regression coefficients with correlated (or longitudinal) data, a sandwich estimator of the covariance matrix may be used to provide standard errors for the estimated coefficients that are robust in the sense that they remain consistent under misspecification of the covariance structure. With large matrices, however, the inefficiency of the sandwich estimator becomes worrisome. We consider here two general shrinkage approaches to estimating the covariance matrix and regression coefficients. The first involves shrinking the eigenvalues of the unstructured ML or REML estimator. The second involves shrinking an unstructured estimator toward a structured estimator. For both cases, the data determine the amount of shrinkage. These estimators are consistent and give consistent and asymptotically efficient estimates for regression coefficients. Simulations show the improved operating characteristics of the shrinkage estimators of the covariance matrix and the regression coefficients in finite samples. The final estimator chosen includes a combination of both shrinkage approaches, i.e., shrinking the eigenvalues and then shrinking toward structure. We illustrate our approach on a sleep EEG study that requires estimation of a 24 x 24 covariance matrix and for which inferences on mean parameters critically depend on the covariance estimator chosen. We recommend making inference using a particular shrinkage estimator that provides a reasonable compromise between structured and unstructured estimators.

Analysis of Variance↗

The ripples of "The Big (agricultural) Bang": the spread of early wheat cultivation.

Demographic expansion and (or) migrations leave their mark in the pattern of DNA polymorphisms of the respective populations. Likewise, the spread of cultural phenomena can be traced by dating archaeological finds and reconstructing their direction and pace. A similar course of events is likely to have taken place following the "Big Bang" of the agricultural spread in the Neolithic Near East from its core area in southeastern Turkey. Thus far, no attempts have been made to track the movement of the founder genetic stocks of the first crop plants from their core area based on the genetic structure of living plants. In this minireview, we re-interpret recent wheat DNA polymorphism data to detect the genetic ripples left by the early wave of advance of Neolithic wheat farming from its core area. This methodology may help to suggest a model charting the spread of the first farming phase prior to the emergence of truly domesticated wheat types (and other such crops), thereby increasing our resolution power in studying this revolutionary period of human cultural, demographic, and social evolution.

Agriculture↗

[Atrial natriuretic factor: one of the mechanisms of action of the phlebology bath at Barbotan].

UNLABELLED: The effect of thermal baths on oedema of the lower limbs might be explained by physical mechanisms of hydrostatic pressure resulting from the use of a deep bath and a centripetal underwater jet, by which the veins and lymph ducts are drained every day. The purpose of this experiment is to demonstrate the existence of hormonal mechanisms which would account for the diuretic effect of thermal baths. One of the effects observed with hydrotherapy is the physiological diuresis that follows each bath, this diuresis would appear to depend at least in part on the atrial natriuretic factor (ANF). The criteria by which assessment can be made essentially biological: ANF level and its biological effects on blood and urine; aldosterone level; plasma renin activity (PRA); creatinine clearance; hematocrit; proteinemia; and blood and urine electrolyte balance. The inclusion criteria are: subjects selected at random and willing cooperate. The criteria for exclusion are disease states which modify ANF kinesis: congestive heart failure, cardiac rhythm disorders, decompensated cirrhosis of liver, obesity, treatment antihypertensive drugs. METHODS: Thirty patients were put through the same experimental sequence, as follows: emptying of the bladder and ingestion of 200 cc of water; seated rest fort 30 mn, after which (to): blood sample; urine sample; ingestion of 200 cc of water; deep bath for 20 mn, i.e. the basic hydro treatment in phlebology at Barbotan. The deep bath is specific to Barbotan and the patient is subjected to maximum immersion in water at a mesothermal temperature of 34.5 degrees C, followed by (t1): blood and urine sample; ingestion of 200 cc of water; supine rest for 90 mn, followed by (t2): blood and urine sample. RESULTS: Data from twenty-eight patients were usable. In this protocol, we use variance analysis with repeated measurements and a 95% confidence limit. The mean value of the principal parameters studies are set out in the following table; these value are accompanied by the degree of significance of the modification at (t1) and (t2). Our experimentation with thirty patients showed that the big thermal bath at Barbotan produces a highly significant increase in ANF secretion, resulting in the diuresis observed after the use of the bath. The antagonist effect of AFN on the renin--angiotensin-aldosterone system was corroborated: we found decreased aldosterone, PRA and creatinine clearance, and increased diuresis and natriuresis. The renal and cardiovascular effects observed after extended immersion in the Barbotan bath (increased diuresis, tachycardia and hypotension, transitory venous vasoplegia and ephemeral vasodilatation of the surface capillaries) are the result of increased ANF secretion. [formula: see text] Supine rest immediately after the bath is essential. This sustains the enhanced ANF and thus reinforces its renal effects, while reducing adverse cardiovascular effects such as the orthostatic hypotension and venous vasoplegia that are normally observed after use of the bath. Moreover, by reducing venular and lymphatic pressure, clinostatism facilitate interstitial to intravascular tissue fluid exchanges and thus helps to drain oedema from the legs. It is striking to note that the hydrotherapy prescribed at Barbotan les Thermes has always included the three most potent factors for ANF release: deep immersion in the big bath, immediate supine rest, and walking. Physiological diuresis has thus been induced empirically as an essential part of the treatment of lower limb phlebopathies.

Aldosterone↗

[Prostate hypertrophy. Surgical experience in Belgium. Multi-center study of 1,140 patients of 80 years and older].

An inquiry organized by the Belgian Society of Surgery brought data on the mortality and morbidity of surgery on patients of 80 years and older for benign prostatic hypertrophy in a total number of 1,140 operations. Consensus exist to see this surgery as planned surgery. The mortality was 73 (6.3%) without statistical difference between endoscopic and open surgery. The morbidity of 824 patients was 165. This last figure relates to the big series in the literature. The mortality is in direct relation to the age of the patients. Evaluation of cardio-pulmonary risk (70.8% of the mortality), operation on indication at earlier diagnosis and prospective studies on the risk of operation in this population group are indicated.

Aged↗

Unexpected death of an infant sibling.

When infants die suddenly and unexpectedly, family structures are abruptly altered. This loss and its subsequent changes affect remaining older siblings. New "big brother" and "big sister" roles are suddenly terminated, often in a catastrophic manner. Young surviving children are sometimes unable to understand the meaning of this event, its impact on the family, and their own role in what has occurred. In this study, 26 families that had sustained the sudden and unexpected death of an infant and that had surviving children were interviewed to obtain data about surviving siblings at least 10 months following the loss. Among the 26 families, there were 35 surviving siblings (ages 16 months to 6 years). The interview schedule sought information relevant to changes in patterns of sleep, toilet training, feeding habits, peer relationships, and parent-child interaction. Among these 35 surviving siblings, parents of 28 siblings (80%) perceived changes in their child's interaction with them, 24 siblings (69%) demonstrated changes in sleep patterns following the baby's death, and 13 siblings (37%) showed changes in social interaction. Regression in toilet training and changes in feeding patterns were infrequent and not areas of major concern for parents. These behavioral changes reflected both a continuum of adjustment by the child and a persistence of parental worries.

Birth Order↗

[Patient satisfaction with the work of the hospital medical personnel].

The article presents the data of patient's satisfaction with the work of nurses and physicians in the hospitals, which belong to Lithuanian Health promotion hospitals network. All nine hospitals of the Lithuanian health promotion hospitals network took part in this study. Altogether 1300 questionnaires were handed out. They have been completed and returned by 1271 patients, i.e. 97.8%. Patients were asked to complete uniform anonymous questionnaire and evaluate the work, behavior and educational activity of nurses and physicians. The majority of respondents assessed the work of nurses and physicians positively, respectively 96.1% and 95.5%. The evaluation of nurses' work depended on patients' sex and the size of the hospital in which they were treated. Male patients and the ones who were treated in big hospitals assessed the work of nurses better than females and patients in smaller hospitals. The evaluation of physicians' work was related to the patients' education and extra payment for health services. Patients, who had higher education and indicated that they had paid additionally, assessed the work of physicians more critically. The majority of patients assessed the behavior of nurses and physicians as very good or excellent, respectively 86.0% and 90.5%. The work of nurses was better evaluated by elderly people and patients treated in big hospitals and as well as in the departments of internal medicine. Patients' satisfaction with the behavior of physicians was associated with patients' education. Better-educated people were more critical in assessing the behavior of physicians. The results of this study have shown that patients' satisfaction depends on many factors. To improve the quality of health care services in hospitals it is necessary to study patients' needs, expectations and factors associated with patients' satisfaction.

Adult↗

Endothelin-converting enzyme activity in human serum lipoprotein fraction.

Endothelin-1 (ET-1)-converting enzyme (ECE) activity in the human serum lipoprotein fraction was studied using a sensitive enzyme immunoassay and reverse-phase high performance liquid chromatography. The ECE activity of cleaving synthetic human big ET-1 into ET-1 by the serum lipoprotein fraction was about 14-times greater than that by whole serum, and the activity was closely associated with lipoprotein itself. The lipoprotein ECE activity, which was optimal at pH 7.0, was inhibited by EDTA, o-phenanthroline, phosphoramidon, thiorphan, phenylmethanesulfonyl fluoride and chymostatin, but not by cysteine or aspartic proteinase inhibitors, suggesting metalloproteinase- and chymotrypsin-like properties. These results suggest that the serum lipoprotein ECE may be involved in the processing of big ET-1 to ET-1 in the circulatory system.

Amino Acid Sequence↗

Substrate selectivity of human cytochrome P450 2C9: importance of residues 476, 365, and 114 in recognition of diclofenac and sulfaphenazole and in mechanism-based inactivation by tienilic acid.

A series of six site-directed mutants of CYP 2C9 were constructed with the aim to better define the amino acid residues that play a critical role in substrate selectivity of CYP 2C9, particularly in three distinctive properties of this enzyme: (i) its selective mechanism-based inactivation by tienilic acid (TA), (ii) its high affinity and hydroxylation regioselectivity toward diclofenac, and (iii) its high affinity for the competitive inhibitor sulfaphenazole (SPA). The S365A mutant exhibited kinetic characteristics for the 5-hydroxylation of TA very similar to those of CYP 2C9; however, this mutant did not undergo any detectable mechanism-based inactivation by TA, which indicates that the OH group of Ser 365 could be the nucleophile forming a covalent bond with an electrophilic metabolite of TA in TA-dependent inactivation of CYP 2C9. The F114I mutant was inactive toward the hydroxylation of diclofenac; moreover, detailed analyses of its interaction with a series of SPA derivatives by difference visible spectroscopy showed that the high affinity of SPA to CYP 2C9 (K(s)=0.4 microM) was completely lost when the phenyl substituent of Phe 114 was replaced with the alkyl group of Ile (K(s)=190+/-20 microM), or when the phenyl substituent of SPA was replaced with a cyclohexyl group (K(s)=120+/-30 microM). However, this cyclohexyl derivative of SPA interacted well with the F114I mutant (K(s)=1.6+/-0.5 microM). At the opposite end, the F94L and F110I mutants showed properties very similar to those of CYP 2C9 toward TA and diclofenac. Finally, the F476I mutant exhibited at least three main differences compared to CYP 2C9: (i) big changes in the k(cat) and K(m) values for TA and diclofenac hydroxylation, (ii) a 37-fold increase of the K(i) value found for the inhibition of CYP 2C9 by SPA, and (iii) a great change in the regioselectivity of diclofenac hydroxylation, the 5-hydroxylation of this substrate by CYP 2C9 F476I exhibiting a k(cat) of 28min(-1). These data indicate that Phe 114 plays an important role in recognition of aromatic substrates of CYP 2C9, presumably via Pi-stacking interactions. They also provide the first experimental evidence showing that Phe 476 plays a crucial role in substrate recognition and hydroxylation by CYP 2C9.

Amino Acids↗

Investigating the construct validity of the dissociative taxon: stability analyses of normal and pathological dissociation.

Research has suggested the existence of a pathological dissociative taxon. However, relatively little is known about this taxon. This study examined the 2-month retest stability of this taxon--together with other measures of dissociation and the Big Five--in a sample of 465 undergraduates. Contrary to expectation, taxon scores were only modestly stable and were substantially less stable than the other measures, including continuous indicators of dissociation. Furthermore, most individuals who were identified as taxon members at one assessment were classified as nonmembers at the other. These results challenge the existence of a pathological dissociative taxon. More generally, these data demonstrate that statistically identified taxa need to be explicated through the normal process of construct validation.

Adult↗

A new scintigraphic method to characterize ultrafiltration in hollow fiber dialyzers.

Ultrafiltration and pressure profiles in hollow fiber dialyzers with different hydraulic permeabilities have been investigated with a new scintigraphic method. Radiolabelled albumin macroaggregates, used as a nondiffusible marker molecule, were added to the blood in an in vitro circuit and circulated through cuprophan and polysulphon dialyzers. Since the marker molecule was too big to cross the dialysis membrane, its changes in concentration were assumed to occur in response to the variation of the blood water content (filtration or back-filtration). These changes in concentration, recorded by a gamma camera, were evaluated to establish the cumulative values of filtration and back-filtration and their relevant profiles along the length of the dialyzer. The achieved data were compared with the experimental values of ultrafiltration empirically measured and with the theoretical values predicted by a classic linear method. Two conditions were analyzed: A) the minimal filtration rate necessary to avoid back-filtration (critical filtration); and B) the condition of zero net filtration in which filtration equals back-filtration. The nuclear method proved to be extremely precise in predicting the ultrafiltration values and significantly more precise than the linear method, especially for the highly permeable dialyzer. The reason for that probably depends on the non-linear pressure and ultrafiltration profile observed with the scintigraphic pattern of the dialyzer. Viscosity changes and local variations in blood flow may in fact interfere with the pressure drop inside the hollow fibers and result in such a complex behavior. The other interesting aspect of this method is the possibility of accurate measurement of the amount of back-filtration that wouldn't be possible with simple calculations. In conclusion, the complex nature of the phenomena regulating the water fluxes in hollow fiber dialyzers requires more complex calculation than a simple linear model to achieve an accurate range of predictability.

Biophysical Phenomena↗

Synthesis of human big endothelin-1 by sequence-specific proteolysis of a fusion protein in Escherichia coli.

Three DNA constructs, pETB-40, 41, and 42, encoding human big endothelin-1 (ET-1) preceded by the specific recognition sequence (Ile-Glu-Gly-Arg) for the activated blood coagulation factor Xa (FXa), fused in frame to the N-terminal portion of beta Gal, were expressed in Escherichia coli. The fusion proteins, pETB-40P, 41P, or 42P, consisted of the 55-, 51-, or 42-aa N-terminal peptide of beta Gal and the 38-aa of big ET-1, and had 1, 0, or 0 Cys residues and 5, 5, or 1 Arg residues in the N-terminal peptide of beta Gal, respectively. Enzymatic cleavage of the purified fusion proteins by FXa or trypsin allowed the recovery of authentic human big ET-1. The rates of conversion of pETB-40P, 41P, and 42P to big ET-1 by FXa digestion were 5.6, 11.2, and 30.0%, respectively. pETB-40P with a deletion of one Cys residue and four Arg residues in the N-terminal part was a better substrate than the other two for FXa or trypsin in the production of big ET-1.

Amino Acid Sequence↗

Cleavage of human big endothelin-1 by Candida albicans aspartic proteinase.

A Candida albicans aspartic proteinase (CAP), one of the secretory proteinases of Candida albicans, is thought to be a possible virulence factor in Candida albicans infection. Whereas endothelin-1 is found as an endothelium-derived strong vasoconstrictive peptide, it is known to have a role in the maintenance of vascular homeostasis and tissue survival. Endothelin-1 is generated from a precursor form of endothelin-1, the so-called big endothelin-1. It has recently been reported that cathepsin D, E and pepsin, which are aspartic proteinases, convert big endothelin-1 to endothelin-1. In this study, the relationship between CAP and big endothelin-1 was studied. High performance liquid chromatography analysis revealed that big endothelin-1 was cleaved into several amino acid sites by CAP, but endothelin-1 was not converted from big endothelin-1. CAP cleaved big endothelin-1 at different sites when compared with that of other known aspartic proteinases, and it suppressed endothelin-1 production through the degradation of big endothelin-1. CAP may break homeostatic mechanism of endothelin-1 in Candida albicans infectious lesions.

Amino Acid Sequence↗

Identification of amino acid residues in the C-terminal tail of big endothelin-1 involved in processing to endothelin-1.

Big endothelin-1 (big ET-1) is converted to 21-amino acid residue endothelin-1 (ET-1) via a specific cleavage at Trp21-Val22 by endothelin converting enzyme (ECE). This conversion is an essential step to produce bioactive ET-1 and represents a regulatory site in the biosynthesis of this potent vasoconstrictor. ECE-1a, a unique membrane-bound enzyme, processes big ET-1 more efficiently than other big ET isoforms, which mainly differ in the C-terminal tail (residues 22-38). In this study, each of the highly conserved residues, Val22, Pro25, Pro30, Gly32, Leu33, and Gly34 were replaced with Ala in the preproendothelin-1 (PPET-1) cDNA using site-directed mutagenesis. The mutant and wild-type cDNAs were transiently transfected into Chinese hamster ovary cells along with ECE-1a cDNAs, and concentrations of the resulting recombinant peptides, ET-1 and big ET-1, in the transfection media were then measured. The concentration of immunoreactive ET-1 in the media from Val22, Pro25, Pro30, Gly32, and Leu33 mutant PPET-1-transfected cells was 4- to 6-fold lower than that of wild type and (Gly34-->Ala)PPET-1. Moreover, with the exception of Gly34 there was a corresponding increase in the concentrations of immunoreactive big ET-1 in the media from mutants. Similar results were obtained when His27, Val28, and Ser35 of big ET-1( )were substituted with the corresponding residues in big ET-2 and big ET-3. These findings suggest that the C-terminal tail has an important role in the intracellular processing of big ET-1 by ECE-1a. Herein we also report that a recombinant big ET-1 analog we previously generated and characterized, (Ala21)big ET-1, inhibits ECE-1a activity in a dose-dependent (K i=1 microM) and competitive manner.

Amino Acid Sequence↗

Childcare immunization programme--to what extent are children covered by vaccinations in Greenland?

OBJECTIVES: New vaccines in the child healthcare programme are regularly considered. When considering the introduction of the Hepatitis-B vaccine into the programme, it was asked to what extent children in Greenland were actually covered by the existing immunization series. Would introducing new vaccines make any sense if the percentage of children immunized was shown to be low? A telephone enquiry conducted in 2000 showed a tendency of very high percentage of full coverage per child (98-99%) in small cities and settlements, as health visitors had a policy of checking up on the immunization status for each child. In big cities and Nuuk, parents alone have the responsibility of having their child immunized. The coverage was a guess of 85-95% for infants, falling as the child grows older. STUDY DESIGN: Retrospective review of registration from all Greenlandic health districts. METHODS: The registration included the number of immunized children in each group of age according to the childcare immunization programme. The collected data were compared to the number of children per age group in each health district. RESULTS AND CONCLUSION: The conclusion must be taken with some reservation on the basis of several aspects of uncertainty. As a whole, the tendencies show reasonably high vaccination coverage countrywide, but variations between districts are great. A number of children probably have their vaccination postponed somewhat in relation to recommended age, but it is fairly evident that the majority of these children are dealt with later. The coverage in a few districts is lower than desired, especially concerning DiTe4 and MMR2.

Adolescent↗

Closure of simple heel ulcers by skin stretching.

A majority of heel ulcers, at least to begin with, extend to dermis or to the fat pad in its superficial part and an appropriate skin closure can heal these ulcers as most of the padding is in tact. Since the skin is adherent to the deeper structures with fibrous bands it has to be stretched or undermined (by cutting the fibrous bands) to close the wound without tension. 17 feet in 11 patients (10 males; one female) in the 12-54 year age-group were operated upon and followed up. Because skin is adherent to deeper tissues by fibrous septae, stretching of skin was planned to mobilize it for a tension-free closure. Of the 17 feet, 13 could be re-examined after 30 months or more. Most of the minor recurrences were seen in the first 6 months after surgery. Major recurrences were seen in 2 feet (one case). The suture line did not show hyperkeratosis and the scar merged well into the surrounding skin after one year. Available data suggest that simple heel ulcers can be made to heal with a good scar by skin-stretching and suture, and, by radiography of the foot, it is worth separating those cases in which ulcer is not extending deep involving calcaneum. The size of the ulcer in heel is important for the success of the operation. The procedure is not intended for big wounds (>15 mm in width).

Adolescent↗