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Biomedical subjects

J Becker

Publications and source records attributed to J Becker.

At least 181 records · Page 10Linked to original sources

Expression of proteins encoded by Epstein-Barr virus trans-activator genes depends on the differentiation of epithelial cells in oral hairy leukoplakia.

The Epstein-Barr virus (EBV) immediate early gene product BZLF1 was localized by indirect immunofluorescence to the cytoplasm of the basal epithelial layer at the lateral border and dorsum of tongue in human immunodeficiency virus-infected and -seronegative patients. Two biopsies of oral hairy leukoplakia revealed a sporadic cytoplasmic staining of the BHRF1 and BRLF1 gene products in the basal epithelial layer. The widespread presence of BZLF1 in the basal epithelial layer indicated that this cell layer contained EBV DNA and was probably directly infected by EBV. Nuclear localization of the immediate early and early gene products BZLF1, BHRF1, BRLF1, and BMLF1 was limited to oral hairy leukoplakia in human immunodeficiency virus-seropositive patients and revealed a codistribution with the virus capsid antigen. Our results indicate that the epithelium of the tongue is a potential reservoir for EBV and that in heavily immunocompromised patients EBV may move from the cytoplasm to the nucleus with increasing differentiation and be coactivated there during the terminal differentiation of epithelial cells at the lateral border and dorsum of tongue.

Cell Differentiation↗

Comparative investigation of c-erbB2/neu expression in head and neck tumors and mammary cancer.

Normal tissues, primary tumors, and metastases of mammary and salivary glands and oral/laryngeal mucosa have been analyzed with Northern-blots employing 32P-labeled RNA probes for the expression of the neu oncogene. Neu oncogene expression of a mRNA species of 4.6 kilobases was found in all normal salivary (five) and mammary glands (four) as well as in two normal or inflamed samples of tongue mucosa. This expression was regarded as baseline activity of the neu gene for the respective tissues and was used as standard for the evaluation of benign and malignant tumors. None of 14 squamous cell carcinomas of the oral and laryngeal mucosa showed enhanced neu transcription level. Five fibroadenomas, one benign variant of phylloid tumor, one carcinosarcoma, and one of two proliferative fibrocystic diseases of the breast showed lacking or normal baseline expression of the neu oncogene, as did one monomorphous cystadenolymphoma of the parotid gland. In contrast, four parotid pleomorphic adenomas and one salivary gland adenocarcinoma showed enhanced neu expression. For mammary adenocarcinomas, increased neu oncogene expression concerned ten of 34 cases--all being variants of ductal carcinomas--and all metastases analyzed (six) deriving from three primaries. One adenoid cystic carcinoma also showed enhanced neu expression. Neu overexpression may reflect accidents of genomic reconstitutional events occurring regularly within the differentiation pathway of epithelial/myoepithelial cells. This assumption was supported by further immunohistochemical analysis which showed stainings of myoepithelial and myoepithelia-like cell populations in tumors, especially pleomorphic adenomas and adjacent normal-looking tissues.

Adolescent↗

Localization of malignant melanoma using monoclonal antibodies.

Finding a screening test to evaluate patients with cancer for occult metastatic disease, as well as imaging all known disease, is a goal of research efforts. Twenty-nine evaluable patients with deeply invasive (stage I), regional nodal (stage II), or systemic (stage III) melanoma underwent imaging by administration of a preparation of the antimelanoma antibody labeled with technetium 99m. Scan results indicated that 28 of 32 confirmed metastatic sites were imaged with this technique (88% sensitivity). Analysis of the individual positive sites revealed that nodal basins and visceral metastases accounted for the highest percentage of metastatic sites imaged, with 14 (88%) of 16 nodal basin metastases and all four visceral metastases being detected through imaging. Occult nodal disease was detected in the iliac nodal chain in two of the 29 patients. The imaging of benign tumors and nodal basins not containing disease accounted for a confirmed false-positive rate of 21%. Three (10%) of the 29 scan results were confirmed to be false-negative. In vivo tumor localization with monoclonal antibodies showed a sensitivity similar to that of other roentgenographic procedures for identifying metastatic disease and was useful in two of three patients in identifying occult iliac nodal disease, a region that is difficult to evaluate with physical examination and other imaging modalities.

Adult↗

Detection of submicroscopic lymph node metastases in patients with melanoma.

A cell culture technique was developed to investigate submicroscopic lymph node metastases in patients with stage 1 or 2 malignant melanoma. Lymph nodes were isolated from standard dissections and bivalved. Half of the node was evaluated by routine histopathologic examination, while the other half was processed and placed into tissue culture. Three hundred twenty-three lymph nodes were collected from 41 patients. The cell culture technique identified 155 of 323 lymph nodes containing micrometastases, while only 20 of 323 lymph nodes tested positive with routine histochemical processing. Nine patients were upgraded from stage 1 or 2 to stage 3 disease after micrometastases were identified in lymph node cultures. Identification of melanoma was confirmed by cytologic examination, immunohistologic staining, and the presence of GD3 ganglioside and 250-kd glycoprotein melanoma-associated antigens. This study provides evidence that the culture of lymph nodes is a sensitive method for the detection of micrometastases. In addition, this procedure may change prognosis and identify candidates for adjuvant therapies.

Antibodies, Neoplasm↗

Mutational analysis of the putative effector domain of the GTP-binding Ypt1 protein in yeast suggests specific regulation by a novel GAP activity.

Ypt1p of Saccharomyces cerevisiae is a ras-related GTP-binding protein that fulfils an essential function in intracellular protein transport between the endoplasmic reticulum (ER) and the Golgi complex. Ypt proteins from yeasts and mammals that share an identical sequence in the region analogous to the ras effector domain are functionally interchangeable. We analyzed the function of the putative effector domain of yeast Ypt1p (amino acids 37-45) using site-directed mutagenesis and gene replacement. Four out of six point mutations leading to single amino acid substitutions (Y37F, S39A, T40S and V43E) did not cause any particular phenotype. ypt1(I41M) mutants were inviable whereas ypt1(D44N) mutant cells were temperature sensitive at 37 degrees C and accumulated core-glycosylated invertase at the nonpermissive temperature. This mutant also accumulated ER and small vesicles both at 25 degrees C and 37 degrees C. From porcine liver we identified and partially purified a GTPase-activating protein (yptGAP) that is similarly active with mouse ypt1p/rab1p and yeast Ypt1p but is inactive with H-ras protein as a substrate. Although none of the yeast ypt1 mutant proteins were significantly impaired in their ability to bind GTP, purified ypt1(D44N)p responded only partially and ypt1(I41M)p did not respond at all, to yptGAP. Thus we suggest that analogous to rasGAP/H-ras p21 interaction in mammalian cells, yptGAP is an intracellular target of Ypt1p, interacting with the effector domain and regulating its GTPase activity, and that this interaction is required for the functioning of yeast Ypt1p in intracellular protein transport.

Amino Acid Sequence↗

Immunoelectron microscopy shows an atypical pattern and a quantitative shift of collagens type I, III and VI in oral Kaposi's sarcoma of AIDS.

The localization of collagen types I, III and VI in normal human alveolar and palatal mucosa and in oral Kaposi's sarcoma (KS) was studied by light microscopy and cryo-immunoelectron microscopy. Normal oral mucosa revealed two different types of organization. The upper connective tissue stroma contained a loose reticular network mainly composed of collagen types III and VI, while collagen type I immunostaining predominated in the deeper stroma. Ultrastructurally, in the KS tumour stroma, a loose pattern of individual thin collagen fibrils was noted. These often fanned out at their ends showing a filamentous substructure. The fibrils consisted predominantly of collagen type I similar to individual fibrils of normal oral mucosa. However, there was a marked loss of thick fibre bundles of collagen types I and III in KS compared with normal oral mucosa, whereas collagen type VI was markedly increased and found preferentially in clusters and strands around cross-striated fibrils that often spanned the distance between single collagen fibres. The abundance of collagen type VI in a pattern similar to early stages of wound healing suggests that the KS stroma resembles an early organizational stage of the interstitial and vascular extracellular matrix subject to a high rate of collagen turnover. This character of the KS stroma appears to result from a continuous auto-and paracrine stimulation of cell growth and collagen synthesis and provides an excellent model to study the structural arrangement of collagen type VI in relation to the fibrillar collagen types I and III.

Acquired Immunodeficiency Syndrome↗

Cardiohemodynamic and serum catecholamine response to surgical removal of impacted mandibular third molars under local anesthesia: a randomized double-blind parallel group and crossover study.

The aim of this study was to differentiate between the extent to which surgical stress and the epinephrine in local anesthetic solutions influence serum catecholamine, cAMP, and potassium levels, and contribute to changes in cardiohemodynamic parameters. One hundred sixty mg of articaine hydrochloride (4.0 mL of a 4% articaine hydrochloride solution) with two different epinephrine doses was injected into outpatients prior to removal of an impacted mandibular third molar in a randomized, double-blind parallel group and crossover design. The results showed that the amount of epinephrine absorbed from the intraoral injection site predominantly determined the serum epinephrine concentration. The anesthetic-induced increase in the serum epinephrine level did not correlate with changes in the cardiohemodynamic parameters under study at any time during the operative procedure. The serum cAMP changes correlated with those of epinephrine, whereas the serum potassium levels remained unchanged. The procedure of tooth extraction was a stressful event when the 1:200,000 epinephrine-containing anesthetic solution was used, showing that the risk of inducing a cardiovascular incident during oral surgery seems to be higher the greater the extent of operation and the lower the epinephrine dose in the anesthetic solution.

Adult↗

[Thiopental concentrations in the whole blood of pregnant and non-pregnant patients in the first minutes following injection. Forensic aspects].

Thiopentone concentrations in blood following injection of the induction solution (Thiopental, 2.5%) may be critical in the context of lethal complications arising during induction of anaesthesia and thereby the toxicological aspects of this drug can become an issue in forensic considerations. The study was designed to find out whether the dose of injected thiopentone may be calculated from blood levels measured in nine pregnant and ten non-pregnant patients. The doses injected ranged from 4-5 mg per kg body weight. The drug was administered as a single bolus dose to the patients undergoing Caesarean section. In the non-pregnant group the first dose of thiopentone was given over 25 s. After that repeated doses of 50 mg were injected over 2s every 20s until the eyelid reflex was ceased. Blood samples for determination of thiopentone were withdrawn from an indwelling catheter inserted to a large cubital vein in the arm opposite that used for the injection and thiopentone levels were measured from blood after column extraction (Extrelut) using spectrophotometry. Venous peak values were found 40-115s after injection in the Caesarean group (mean: 20.3 micrograms kg-1) and were higher than in the non-pregnant patients (97-190s, mean: 17.8 micrograms kg-1). No correlation was found regarding venous peak levels (mgkg-1 blood) and induction dose (mgkg-1 body weight), neither in the pregnant nor in the non-pregnant group. Our findings would suggest that the injected thiopentone dose cannot be calculated from the concentration of the drug in the blood. Forensic conclusions concerning the onset of lethal complications during the induction of anaesthesia cannot be drawn on the grounds of venous thiopentone concentrations.

Adult↗

The screen for caregiver burden.

The Screen for Caregiver Burden (SCB) is a measure designed to identify distressing caregiver experiences rapidly. The 25-item SCB, created specifically for spouse caregivers of Alzheimer's care recipients, provides scores for objective burden (OB; number of potentially negative experiences) and subjective burden (SB; appraised distress in response to the experiences). Three separate studies (total N = 191) demonstrate the internal consistency, test-retest reliability, and validity (content, divergent, convergent, and criterion) of the scales. Finally, evidence suggests that changes in OB and SB over time are explained by changes in care-recipient functioning and caregiver distress.

Aged↗

Distribution of collagens type V and VI in the normal human alveolar mucosa: an immunoelectronmicroscopic study using ultrathin frozen sections.

The ultrastructural localization of collagens type V and VI in normal human gingival mucosa was investigated by immunoelectron microscopy. Twenty biopsies were fixed in dimethylsuberimidate and shock-frozen in slush nitrogen. Collagen type V was mainly located to meshworks of uniform nonstriated microfibrils of 12 to 20 nm width, which preferentially appeared in larger spaces between cross-striated major collagen fibrils. Occasionally single microfibrils of collagen type V fanned out from the ends of major collagen fibrils, which may indicate a role as a core fibril. Collagen type V was not found in the subepithelial basement membrane and the immediately adjacent stroma. Collagen type VI was detected in a loose reticular network of unbanded microfilaments that were morphologically distinguishable by knoblike protrusions every 100-110 nm. These microfilaments were found in the vicinity, but not as an intrinsic component, of the subepithelial basement membrane. Single filaments of collagen type VI filaments appeared to form bridges between neighboring cross-striated major collagen fibrils, suggesting an interconnecting role for this collagen type. The method presented appears to be excellently suited to study the normal and pathological supramolecular organization of the oral extracellular matrix.

Adolescent↗

Immunoelectron microscopic localization of collagens type I, V, VI and of procollagen type III in human periodontal ligament and cementum.

We examined the ultrastructural localization of collagens Type I, V, VI and of procollagen Type III in decalcified and prefixed specimens of the periodontal ligament and cementum, by immunoelectron microscopy using ultra-thin cryostat sections. Immunostaining for collagen Type I was pronounced on the major cross-striated fibrils entering cementum and in cementum proper, whereas staining for procollagen Type III was almost exclusively observed on the major fibrils in the periodontal ligament situated more remote from cementum. Reactivity for collagen Type V was limited to aggregated, unbanded filamentous material of about 12 nm diameter that was found mainly in larger spaces between bundles of cross-striated collagen fibrils and occasionally on single microfibrils that apparently originated from the ends of the major collagen fibrils, which may support the concept of this collagen as a component of core fibrils. Collagen Type VI was present as microfilaments appearing to interconnect single cross-striated fibrils. In the densely packed fibril bundles of the periodontal ligament, no collagen type VI was detected. Neither Type V or Type VI collagen was observed in cementum.

Adolescent↗

EEG and skeletal development in children with different psychosocial characteristics.

Two groups of children with different socioeconomic level were studied. One minute EEG at rest was recorded in monopolar leads F3, F4, C3, C4, P3, P4, O1, O2, F7, F8, T3, T4, T5 and T6. Absolute and relative power in four EEg bands (delta, theta, alpha and beta) were computed. Radiographies of the left hand and the wrist were also obtained in all children. Age regression equations of the variables derived from EEG spectra were calculated in each group. In the group with low socioeconomic level many children had antecedents of risk factors. In this group absolute and relative power in the four bands presented a great dispersion and no correlation with age. In the group with good socioeconomic level the age regression equations of the EEG variables were significant, absolute values in the four bands decreased with age, as well as delta and theta relative power, while alpha and beta relative power increased with age. The area of the ossification center of each bone of the hand of the lower end of the ulna and radius were obtained from the X-ray film. Linear regression equations for the area of each ossification center were significant in both groups. No intercept or slope differences existed between both groups in any area. It is concluded that psychosocial disadvantage and antecedents of risk factors, although not producing any effect on skeletal development, do affect EEG maturation.

Adolescent↗

On the mechanism of action of H2O2 in the cellular stress.

We propose a hypothesis according to which the reactive and reduced species of oxygen could be the intracellular inducers of the stress (or "heat-shock") response. This hypothesis is based on the following observations on Drosophila cells: a) the return to normoxia after 24 h anaerobiosis is sufficient to induce the synthesis of the "heat shock" proteins without elevation of temperature together with a rapid increase of O2 consumption; b) hydrogen peroxide introduced in the culture medium induces the early transcriptional activation of the "heat shock" genes (maximal after 5 minutes); c) hydrogen peroxide added to cellular extracts in vitro (thus acting as an intracellular metabolite) activates instantaneously the binding capacity of a "heat shock" factor to a DNA "heat shock" regulatory element. Thus, hydrogen peroxide, and possibly other reactive reduced species of oxygen, could trigger the onset of the stress (or "heat shock") response.

Animals↗

An improved technique for the in situ detection of DNA after polymerase chain reaction amplification.

In situ detection of polymerase chain reaction (PCR)-amplified DNA in cell and tissue preparations previously required 5 to 7 primer pairs designed to generate a long (greater than 1,000 base pair) product. The authors describe a nonisotopic PCR in situ technique, employing a single primer pair and target sequences as short as 115 base pairs, that can detect one target molecule per cell. The essential procedural change is to withhold the DNA polymerase or primers until the reaction temperature approaches 80 degrees C. The Hot-Start method greatly increased amplification specificity which, more than product size, appears to determine success of in situ PCR. The marked improvement in specificity may permit target detection by direct incorporation of labeled nucleotides.

DNA, Viral↗

Ecdysterone decreases the transcription level of the retrotransposons 1731 and 412 in a Drosophila cell line.

Cultured Drosophila melanogaster cells are responsive to the steroid hormone 20-hydroxyecdysone (ecdysterone). The transcripts of two copia-like transposable elements, 412 and 1731 were examined in cells treated for different lengths of time by the hormone. Using dot-blot and Northern-blot analysis, we have shown that the transcripts of these two retrotransposons were rapidly decreased by ecdysterone treatment.

Animals↗

Combined Fontana-Masson-mucin staining of Cryptococcus neoformans.

Cryptococci react positively with various histochemical stains, including the Fontana-Masson (FM), which stains the cell wall, and mucin stains, such as alcian blue and mucicarmine, which stain the capsule. Combinations of the FM stain with both the alcian blue and mucicarmine stains were performed on paraffin-embedded tissue specimens that were obtained from 15 patients who had culture-proved cryptococcosis. Combined FM-mucicarmine and FM-alcian blue stains were compared with other individual fungal stains. The FM stain, followed by either the mucicarmine or alcian blue stain, distinctively demonstrated both the cell wall and capsule of most organisms. More organisms were recognized in the combined stains than with either stain done individually. No interference between the stains was noted. Combining the FM stain with either of these two mucin stains appears to be helpful for identifying cryptococci.

Alcian Blue↗

HIV-1 seropositivity and behavioral and sociological risks among homosexual and bisexual men in six Mexican cities.

We report on the epidemiology of sexual behaviors, HIV-1 seroprevalence, and condom use in gay and bisexual men in six Mexican cities in 1988 and test the extent to which variations in sexual behavior and sociological risks are responsible for variations in prevalence. Seroprevalence rates among samples in six cities ranged from 2% to 25%. In multiple logistic regression models controlling for city, insertive/receptive behavior (IRB), and meeting partners in bathhouses, the following were independently related to seropositivity (p less than 0.05): city, IRB, syphilis, sex with a person with AIDS, and meeting partners in bathouses. The independent risks associated with categories of IRB were no or almost no activity (odds ratio 1.0), only insertive (3.0), mostly insertive (4.9), mixed (6.0), mostly receptive (3.3), and only receptive (0.9). The condom use rate on last sexual encounter was 30%. The main sexual risk for HIV-1 infection is not exclusively receptive anal sex, but rather mixed behavior. This association may be explained by the infectious state of the partner pools. The sociologic risk variables (national and local partner pools) are better predictors of seroprevalence than behavioral variables, such as the number of partners or use of condoms. These results imply that more effective individual strategies for risk reduction are needed, including better knowledge concerning the risk status of partners.

Adolescent↗