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At least 613 records · Page 34Linked to original sources
PubMed2000

The enhanced green fluorescent protein as a tool for the analysis of protein dynamics and localization: local fluorescence study at the single-molecule level.

The green fluorescent protein (GFP) has emerged, in recent years, as a powerful reporter molecule for monitoring gene expression, protein localization and protein-protein interaction. Several mutant variants are now available differing in absorption, emission spectra and quantum yield. Here we present a detailed study of the fluorescence properties of the Phe-64-->Leu, Ser-65-->Thr mutant down to the single molecule level in order to assess its use in quantitative fluorescence microscopy and single-protein trafficking. This enhanced GFP (EGFP) is being used extensively as it offers higher-intensity emission after blue-light excitation with respect to wild-type GFP. By means of fluorescence spectroscopy we demonstrate the absence of the neutral form of the chromophore and the lack of photobleaching recovery after ultraviolet light irradiation. Furthermore, we show that the EGFP spectral properties from isolated to densely packed molecules are highly conserved. From these experiments EGFP emerges as an ideal molecule for quantitative studies of intra and intercellular tagged-protein dynamics and fluorescence-activated cell sorting, but not for monitoring single-protein trafficking over extended periods of time.

Base Sequence↗
PubMed1980

Immunohistochemical localization of aldose reductase. I. Enzyme purification and antibody preparation--localization in peripheral nerve, artery, and testis.

Aldose reductase (AR) was purified from rat and bovine seminal vesicles using DEAE-cellulose, hydroxylapatite, and Sephadex-gel column chromatography. The purification resulted in the obtention of an AR pool and a contaminating pool. Antibodies were raised in rabbits against both enzymes by subcutaneous injection of the AR pool. The antisera was judged to be specific for AR by immunoprecipitation of AR activity and by Ouchterlony double immunodiffusion and immunoelectrophoretic methods. Antibodies against rat AR were used in the unlabeled antibody-enzyme (PAP) technique to demonstrate the cellular location of the enzyme in a number of tissues known to be sites of diabetic lesions. Antibodies against bovine AR were not cross reactive with the rat enzyme, as determined by Ouchterlony and competitive protein-binding studies. AR was localized in rat tissues to the Schwann cell sheath of peripheral nerve, arterial endothelium, and the sustentacular (Sertoli) cells and mature spermatids of the testis.

Aldehyde Reductase↗
PubMed2006

Making COD statistics useful for public health at local level in the city of Cape Town: utilidad para la salud pública a nivel local en Ciudad del Cabo.

OBJECTIVE: To review the quality of the coding of the cause of death (COD) statistics and assess the mortality information needs of the City of Cape Town. METHODS: Using an action research approach, a study was set up to investigate the quality of COD information, the accuracy of COD coding and consistency of coding practices in the larger health subdistricts. Mortality information needs and the best way of presenting the statistics to assist health managers were explored. FINDINGS: Useful information was contained in 75% of death certificates, but nearly 60% had only a single cause certified; 55% of forms were coded accurately. Disagreement was mainly because routine coders coded the immediate instead of the underlying COD. An abridged classification of COD, based on causes of public health importance, prevalent causes and selected combinations of diseases was implemented with training on underlying cause. Analysis of the 2001 data identified the leading causes of death and premature mortality and illustrated striking differences in the disease burden and profile between health subdistricts. CONCLUSION: Action research is particularly useful for improving information systems and revealed the need to standardize the coding practice to identify underlying cause. The specificity of the full ICD classification is beyond the level of detail on the death certificates currently available. An abridged classification for coding provides a practical tool appropriate for local level public health surveillance. Attention to the presentation of COD statistics is important to enable the data to inform decision-makers.

Cause of Death↗
PubMed1995

Chalcone synthase localization in early stages of plant development. I. Immunohistochemical use of plasmolysis for localizing the enzyme in epidermal cell cytoplasm of illuminated buckwheat hypocotyls.

Immunohistochemical methods combined with progressive plasmolysis were used to localize chalcone synthase (CHS), an important enzyme for plant metabolism of aromatics in hypocotyls of illuminated buckwheat (Fagopyrum esculentum M.) seedlings. Illumination of etiolated seedlings with white light results in anthocyanin synthesis in the epidermal layer of the hypocotyl. Anthocyanin-containing epidermal peels, after fixation for 30 min in 4% paraformaldehyde, 2.5% glutaraldehyde, 0.1% caffeine, were treated with a specific rabbit anti-buckwheat CHS antibody and a 20 nm goat anti-rabbit IgG gold conjugate. CHS is specifically shown in epidermal cells as pink to dark red deposits. Progressive plasmolysis combined with our immunohistochemical method showed that CHS was located exclusively in the cytoplasm of the epidermal cells of buckwheat hypocotyls except for the guard cells, which contained no detectable CHS.

Acyltransferases↗
PubMedDate not supplied

Early local steroid therapy of infantile eyelid hemangiomas (local steroid therapy of lid hemangiomas).

Two cases of upper eyelid capillary hemangiomas were successfully treated by local injection of steroids. Both cases required two injections, performed between 11 and 20 weeks of age. No untoward side effects occurred. This treatment would appear to be valuable in preventing the development of ophthalmic sequelae commonly associated with benign observation.

Dexamethasone↗
PubMed1999

Regional localization of a gene for nonspecific XLMR to Xp11.3-p11. 23 (MRX51) and tentative localization of an MRX gene to Xq23-q26.1.

Two families with nonspecific X-linked mental retardation (MRX) are presented. In the first family, MRX51, three male patients showed mild to borderline mental retardation. Multipoint linkage analysis yielded a maximal LOD score of 2.10 between markers DXS8012 and DXS1003, localizing the MRX51 gene at Xp11.3-p11.23. In the second family, XLMR7, three men showed moderate mental retardation (MR), and one possible female carrier had mild MR. Multipoint linkage analysis yielded an LOD score of 1.80 between markers DXS8063 and DXS1047, situating the disease gene at Xq23-q26.1. When the analysis was performed considering the affected female to be an expressing heterozygote carrier of the disease mutation, a maximal LOD score of 2.10 was found in the same region.

Chromosome Mapping↗
PubMed2000

[Electron immunohistochemical analysis of localization of neutral Mn2+-dependent DNAase. II. Analysis of ultrastructural localization in epon sections of different organs of the rat].

The use of a conjugate of colloidal gold with monotypic antibodies against a neutral Mn(2+)-dependent DNAse has revealed the enzyme localization in ultrathin Epon sections of glutaraldehyde fixed tissues. Technical procedures involved in fixation, embedding, and immune reactions are described. DNAse has been established in the nuclei in both normal and regenerating liver. A protein, immunologically close to DNAse, has been identified in the nuclei of a lymph node, thymus, spleen and cerebellar cortex.

Animals↗
PubMed2000

[Treatment of localized prostatic cancer--different alternatives. Patients with localized prostatic cancer must actively participate in the choice of therapy].

The treatment options currently available for patients with clinically localized prostate cancer are radical prostatectomy, irradiation and deferred symptom-guided treatment. Today we do not have any controlled data indicating that any of these treatments is better than the other. Management policies that have evolved are, at best, based on structured comparisons of data from uncontrolled treatment series. In such comparisons the differences between the various treatment options with respect to disease-specific survival up to 10 to 15 years after diagnosis are modest. These comparisons are based on patient series diagnosed 15 to 20 years ago, i.e. before prostate specific antigen (PSA) was available as a marker for prostate cancer. The utilization of PSA has dramatically changed the conditions for diagnosing prostate cancer, in that with PSA we find more men with prostate cancer with smaller tumors at diagnosis. Today we find many tumors because of an elevated PSA followed by systematic biopsies in men without palpable prostatic lesions. Thus it is difficult to translate long-term outcome data from older patient series to the current situation in which tumors are found under changed clinical conditions. Consequently, a patient can only be informed about the different treatment options and their side effects. In the treatment decision process he has to balance a possible benefit in survival against side effects, according to his own preferences. We need randomized studies comparing the different treatment options. Such research is under way in Scandinavia, but due to the long course of the disease we will probably have to wait many years before we can answer the questions which sparked these studies.

Antineoplastic Agents↗
PubMed2003

Local problems, local solutions: improving tuberculosis control at the district level in Malawi.

OBJECTIVE: To examine the causes of a low cure rate at the district level of a tuberculosis (TB) control programme and to formulate, implement, and evaluate an intervention to improve the situation. METHODS: The study setting was Mzuzu (population 60,000), where the annual smear-positive pulmonary TB incidence was 160 per 100,000 and the human immunodeficiency virus (HIV) seroprevalence was 67% among TB patients. There is one TB treatment unit, but several other organizations are involved with TB control. An examination of case-holding activities was carried out, potential areas for improvement were identified, and interventions performed. FINDINGS: In 1990-91, the cure rate was 24% among smear-positive cases (29% among survivors to end of treatment). Problems identified included a fragmented TB control programme; inadequate training and supervision; suboptimal recording of patients' addresses; and nonadherence to national TB control programme protocols. These problems were addressed, and in 1992-93 the cure rate rose to 68% (relative risk (RR) = 2.85 (95% confidence interval (CI) = 1.63, 4.96)) and to 92% among survivors to the end of treatment (RR = 3.12 (95% CI = 1.84, 5.29)). High cure rates are therefore achievable despite high HIV prevalence. CONCLUSIONS: Simple, inexpensive, local programmatic interventions can dramatically improve TB case holding. This study demonstrates the need for evaluation, training, and supervision at all levels of the programme.

Antitubercular Agents↗
PubMed2003

Localization of neuropeptide Y and norepinephrine in the porcine ovarian artery and their influence on the local blood pressure.

The aim of the present study was to determine: (i) the presence of dopamine-beta-hydroxylase (DbetaH)- and neuropeptide Y (NPY)-immunoreactive (IR) nerve fibres in the wall of the porcine ovarian artery, (ii) the influence of NPY and norepinephrine (NE) on the contractile activity of the pig ovarian arteries, and (iii) the pharmacological analysis of the interaction between NPY and NE in the isolated porcine ovarian arteries collected from immature pigs and from animals in different days of the estrous cycle. Ovarian arteries for immunohistochemistry and isolated arteries for pharmacological studies were excised from immature pigs and mature animals on days 1-5, 8-13 and 17-20 of the estrous cycle. The study showed that both DbetaH- and NPY-IR nerve fibres were present in the pig ovarian arteries in all periods examined, and, that in some fibres DbetaH and NPY were co-localized. Both NE (10(-6) M) and NPY (10(-7) M) increased blood pressure of examined preparations, however, NE caused stronger changes in the vessel wall tension (P<0.001), than did NPY. NE significantly increased (P<0.001) blood pressure of all isolated arteries, however, this response was stronger in vessels from days 1-5 of the cycle, when compared to days 8-13 (P<0.01), 17-20 and immature pigs (P<0.001). NPY increased significantly blood pressure in isolated arteries from days 8-13 and 17-20 of the cycle (P<0.001), while in preparations taken from immature pigs and animals on days 1-5 of the estrous cycle this response was somewhat weaker (P<0.01). A higher elevation (P<0.001) of blood pressure after NPY administration was observed in isolated arteries from days 17-20 of the cycle, when compared to vessels from days 1-5 and 8-13 and those from immature pigs. Moreover, NE significantly intensified (P<0.001) an increase in the blood pressure in isolated arteries pre-treated with NPY in all periods examined. NPY insignificantly (P>0.05) potentiated increase of blood pressure in NE pre-treated vessels of immature pigs and in isolated arteries from days 17-20 of the cycle and significantly (P<0.05) in vessels from days 1-5 and 8-13 of the estrous cycle. Our results indicate that DbetaH- and NPY-IR nerve fibres are present in the pig ovarian arteries. NE and NPY administered alone increased blood pressure in the pig isolated ovarian artery and simultaneous administration of both substances caused each other potentiation of vasocontractile effect, however, the strength of observed changes was dependent on the stage of the estrous cycle.

Aging↗
PubMed1991

[Circulating prostatic-specific antigen in benign hypertrophy and localized prostate cancer: can PSA be considered a screening examination for localized cancer?].

Prostate-specific antigen (PSA) is increasingly used in the diagnosis of prostatic pathology. Its usefulness in the early diagnosis of prostatic cancer is controversial. The aim of the study is to evaluate the sensitivity and specificity of PSA in a population with prostate diseases. Moreover, we wanted to know if the measure of the prostate volume may increase the sensitivity of the test. In benign prostatic hypertrophy (88 patients), a good correlation exists between circulating PSA and the prostatic volume or the volume of the adenoma. This correlation disappears in the presence of an adenocarcinoma at the profit of the tumor volume (46 patients). Used as a means of screening for cancer, the serum level of PSA with a threshold value of 2.5 ng/ml has a sensitivity of 91% and a specificity of 32%. The sensitivity is 50% and the specificity is 85% at a level of 15 ng/ml. Taken alone, the level of PSA is inadequate for diagnosis: If the lower level is chosen (2.5 ng/ml), the majority of benign prostatic hypertrophies will be the object of a biopsy. If the higher level is chosen (15-23 ng/ml), 50% of localized cancers of the prostate will escape detection. Nevertheless, a level of PSA < 15 ng/ml is an argument for a strong suspicion in favor of an adenocarcinoma of the prostate. The capacity of BPH to "secrete" serum PSA is five times greater than that of the normal peripheral prostate, and the capacity of cancer is 20 times greater than that of an adenoma. The individual variability of serum PSA per cubic centimeter of prostatic tissue is too great to allow a precise interpretation as a function of volume.

Adenocarcinoma↗
PubMed1991

[Status of local excision of local deep rectal cancers with curative intention].

103 cases have been analyzed retrospectively to define the indication for local extirpation of small and deep-seated rectum carcinomas with curative purpose. While pedunculated carcinomatous adenomas can be removed by endoscopic loop extraction, the non-sessile polypous carcinomas have to be radically extirpated by excision from intestinal wall resection is a promising procedure. Here, the transanal access by means of a funnel-shaped, lateral fenestrated glass speculum as introduced by Dewey yields better results than the rectotomia posterior.

Adult↗
PubMed1990

[New combined therapy of radiation and local administration of OK-432 for esophageal cancer--local control rate and two-year survival rate].

We devised new combination therapy of radiation and local administration of OK-432 and performed for 26 patients with esophageal cancer between March, 1987 and December, 1988 as a pilot study. The average age was 73 years. Among 26 patients, male were 20 and female were six. Patients were irradiated at the schedule of under 2 Gy/day and the dose of TDF (time, dose and fractionation factor) 100 totally. OK-432, 10KE was endoscopically administered around cancer lesion at the beginning of the radiotherapy, and two weeks later, 5KE was given in the same manner. Complete response was obtained 20 out of 26 cases (77%) and partial response was obtained in the remaining six cases (23%). All six patients with tumor length less than five cm showed complete response. All 16 patients who could not eat food orally before treatment, became to take food enough orally after the treatment and could discharge in good condition. One year and two years survival rate of 26 patients by Kaplan Meier method were 67.4% and 47.2%, respectively. Six patients with tumor length less than five cm, are all alive without a sign of recurrence. This combination therapy will improve not only the response rate and the survival rate, but also the quality of life of patients with esophageal cancer.

Aged↗
PubMed1989

[Studies on the mechanism of glomerular localization of immune complexes (1). Localization of preformed native ferritin immune complexes on the glomerular basement membrane in passive serum sickness nephritis of mice].

Immune complex deposits found on the subepithelial side of the glomerular basement membrane (GBM) are characteristic of membranous glomerulonephritis. In order to clarify the mechanism of subepithelial immune complex deposition, we investigated the manner of localization of preformed immune complexes (IC) composed of native ferritin (NF) and anti NF antibody (aNFab) in a 40-fold antigen great excess using the system of passive serum sickness nephritis of mice. The following results were obtained. (1) The ferritin particles in the GBM of animals given NF alone, as controls, were mainly restricted to the lamina rara interna (LRI) with very few penetrating the entire depth of the GBM. (2) In animals given the IC, greater numbers of ferritin particles were seen, in comparison with the control animals, not only in the lamina densa (LD) but also in the lamina rara externa (LRE). (3) Immunoelectron microscopy (IEM) demonstrated that each NF particle was accompanied by aNFab, which was found to be peroxidase positive material, throughout the GBM. (4) On the grounds that ICs were prepared in Ag great excess, it is suggested that ICs seen in the GBM are composed of a low avidity antibody with a low Ag/Ab ratio, probably Ag1Ab1. From these results, it is suggested that circulating ICs are necessary to give rise to IC deposition on the GBM and that NF and aNFab exist together as ICs throughout the GBM. We concluded that subepithelial IC deposits arise from the circulation, and that after GBM trapping, circulating small-sized IC can pass through the LD to reach the subepithelial side of GBM.

Animals↗
PubMed1989

[Studies on the mechanism of glomerular localization of immune complexes (2). Localization of non-dissociable immune complexes on the subepithelial side of the glomerular basement membrane].

In order to investigate whether preformed (circulating) immune complexes (IC) can localize in the subepithelial side of the glomerular basement membrane (GBM), non-dissociable covalently cross-linked IC (cov. IC) composed of native ferritin (NF) and anti NF antibody (aNFab) were preformed in a 2 step reaction using the bifunctional reagent, toluene-2,4-diisocyanate (TDI). Molecular weight of cov. IC was about 58 x 10(4) dalton, which suggests that the cov. IC was composed of Ag1Ab1 lattices. The GBM of mice given cov. IC intravenously showed that deposition of cov. IC was not only in the lamina rara interna but also, with time, in the lamina densa and lamina rara externa. The size of cov. IC and the distribution in the GBM described above were similar to those we studied previously using preformed dissociable NF and aNFab complexes in a 40-fold antigen great excess. From these results, we concluded that preformed immune complex in vitro can pass through the lamina densa as intact IC to give rise to the subepithelial area without dissociation or reformation. That is, the subepithelial IC deposits arise from the trapping of small-sized IC derived from the circulation.

Animals↗
PubMed1986

Localization of cationic proteins derived from platelets and polymorphonuclear neutrophils and local loss of anionic sites in glomeruli of rabbits with experimentally-induced acute serum sickness.

The localization of cationic proteins (CP) derived from platelets and from polymorphonuclear neutrophils (PMN) in glomeruli of 42 rabbits injected i.v. with a large amount of bovine serum albumin, was investigated in sequential biopsies by immunofluorescence, using goat-anti-platelet CP and anti-PMN CP sera. Platelet CP deposits became detectable within 7 to 8 days after the i.v. injection of bovine serum albumin, before or coincident with the onset of proteinuria. The intensity and the extent of linear and segmental deposits of platelet CP along the glomerular capillary walls reached a peak at day 9 to 10, when proteinuria was maximal. The anti PMN-CP serum stained the cytoplasm of the few PMN present in glomeruli and only occasionally at day 11 and 12 identified focal deposits of PMN-CP along the glomerular capillary walls. The kinetic study of glomerular immune deposits showed that the first appearance of immune deposits in antigen excess was preceded by, or was concomitant, with the detection of platelet-CP in glomeruli. In the later stages of serum sickness, the immune deposits showed a progressive increase in rabbit IgG and C3. The glomerular polyanions were studied by light microscopy, using the colloidal iron technique, and by electron microscopy using polyethyleneimine as a cationic probe. The glomerular deposits of platelet-CP were associated with a reduction of colloidal iron staining, which was maximal 9 to 11 days after the i.v. injection of bovine serum albumin. At day 15, colloidal iron staining was almost completely restored. At day 9 in rabbits with acute serum sickness the anionic sites of glomerular basement membrane evidenced by polyethyleneimine, were segmentally decreased, mainly in the lamina rara interna. In rabbit studied at day 15 the anionic sites were decreased only at the base of the subepithelial electron dense deposits (humps). These results suggest that in rabbits with experimentally-induced acute serum sickness, during the early stages of glomerular immune deposit formation endogenous CP, released mainly from platelets, bind to glomerular capillary walls and possibly contribute to the neutralization of glomerular polyanions.

Acute Disease↗
PubMed1986

Formation of the local secondary structure of proteins: local sequence or environment.

An analysis of the occurrence of tetrapeptides in 35 globular proteins for alpha-helix, beta-structure and coil was performed. We concluded that: the conformation of a short polypeptide segment cannot be determined on the basis of the knowledge of the amino acid sequence only; local structures of a protein are formed as the result of interactions within the whole structural domain of the protein as well as interactions with the environment.

Amino Acid Sequence↗
PubMed1985

[Frequency, localization and extent of local wall function disorders of the left ventricle in patients with rheumatic mitral valve diseases].

In 59 patients with rheumatic mitral valve disease, local wall motion was assessed by analysing the diastolic and systolic left ventricular silhouette from biplane angiograms with a quantitative radial axes model using 90 radii in each projection. The left ventricle was dissected into 4 segments in the RAO and 2 segments in the LAO projection and a mean radii shortening for each segment was calculated. 27 patients had pure severe mitral stenosis (MS), 10 patients mitral insufficiency (MI) and 22 patients combined mitral valve disease (MSMI). A mean radii shortening less than 25% was diagnosed as wall motion abnormality. In 78% of all patients wall motion abnormalities could be detected in at least one segment. 32% of patients with MS, 27% of patients with MI and 33% of patients with MSMI had wall motion abnormality of the anteroapical wall. In only 8% of patients with MS and in no patients with MI and MSMI was the posterobasal segment disturbed. The anterobasal and diaphragmal wall motion showed wall motion abnormality in 16% and the septal and the posterolateral area in 17% of all patients. 78% of all patients had at least one diseased segment; motion abnormalities of the anteroapical wall (seg. 2) occurred most often whereas the posterobasal area (seg. 4) was diseased in only 4% of all patients. Ejection fraction was severely impaired (less than 55%) in 7 patients only (5 of these patients had MS) with wall motion abnormalities in more than 4 segments. There was no correlation between the extent of valve disease and occurrence and extent of wall motion abnormalities.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗