The intracellular localization of uteroglobin, a progesterone-induced protein, was studied in uterus and oviduct by means of immunoelectron microscopy with the protein A-gold technique. In the uterus, uteroglobin was synthesized in the columnar epithelium of the endometrium where most of the cells were immunoreactive. The protein was localized mainly in small secretory granules which were seen in the process of release into the uterine lumen. The luminal microvilli were also heavily stained. In the oviduct, the secretory cells contained large immunoreactive granules at the apical zone, some of which were observed while discharging into the lumen. Within these secretory granules, uteroglobin accumulated particularly in lens-shaped patches at the periphery of the granules. In vitro kinetic studies on the secretion of newly synthesized uteroglobin indicated that the ability to store uteroglobin is greater in the oviduct than in the uterus; however, the rate of uteroglobin secretion is greater in the uterus than in the oviduct. Thus, there appears to be a good correlation between the microscopic and the functional observations.

Animals↗

PubMedDate not supplied

[Circumscribed and confluent papular rash in the trunk and hands in a male patient infected with the human immunodeficiency virus].

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AIDS-Related Opportunistic Infections↗

PubMedDate not supplied

[Peculiarities of latex allergy and preventive measures in the pediatric age group].

Latex allergy can be considered a problem of public health in at-risk populations. Although the figures on prevalence vary, approximately 1 % of the general population is sensitized to latex. To date, two determining factors for latex sensitization have been identified: an atopic predisposition and the number of operations. Among atopic patients or those at-risk for allergy, the prevalence of latex sensitization is between 3 and 20 times higher than that among the general population. Nevertheless, the effect of this factor seems to be modulatory and the determining factor seems to be the number of operations that children have undergone; the critical number is 5-6 interventions, after which the probability of sensitization increases considerably.All children who have undergone multiple operations should be considered as being at-risk. Children with spina bifida are especially at-risk. The reasons for this are still a matter of debate: the disease itself has been postulated as a specific risk factor, although other, possible associated factors could be determinant in triggering latex sensitization (the presence of a ventricular-peritoneal shunt, age at which surgery was performed, type of operation, etc.). Children with spina bifida and latex allergy show considerable differences in type of allergic sensitization; these differences seem to result from the different route of sensitization and could explain the variations observed in the clinical manifestations between children and adults: children become sensitized mainly by direct contact between latex particles and blood vessels and open mucosae while in adults the process takes place transcutaneously or by inhalation of aerosol particles. Consequently, the most frequent manifestation in children is urticaria while contact dermatitis and respiratory symptoms predominate in adults. Studies performed with immunoblotting have confirmed that children have IgE that almost constantly recognize low molecular weight latex proteins (14, 15 and 27 Kd) while this finding is less frequent in adults.The preventive measures in the pediatric population focus on avoidance of latex, especially in hospitals. Today, latex is ubiquitous, making complete avoidance difficult. When necessary, and whenever a latex-free environment cannot be guaranteed, presurgical medication can be used, although its utility is debatable. Although further studies are required, specific immunotherapy is one therapeutic possibility that may in future be used in children with latex allergy.

Child↗

PubMedDate not supplied

Adsorption-desorption of antigen to polystyrene plates used in ELISA.

Since ELISA reliability depends to a great extent upon solid-phase reagent concentration and stability, we sought to analyse the influence of experimental conditions during ELISA performance on the adsorption/desorption of proteins to microplates. The effect upon desorption of several experimental parameters (antigen concentration, antibody concentration and affinity, washings, conjugate and inhibitor incubations) and quantitative treatment of protein-polystyrene adsorption were analysed. The adsorption to polystyrene microplates was studied with a hapten-conjugated protein (BSA-Ar36) in order to facilitate the analysis of the influence of antibody affinity on desorption during ELISA. Our results show that polystyrene plates adsorb BSA-Ar36 according to the Langmuir isotherm. The adsorption constant was 2.1 X 10(8) L/mol and maximal surface concentration of protein on solid phase was 1.8 X 10(-7) g/cm2. Although desorption was present, we observed that it did not affect the reliability of results of either direct or inhibition ELISA, because it was not dependent on the composition of the sample.

Adsorption↗

PubMedDate not supplied

Stable high-copy-number integration of Aspergillus oryzae alpha-AMYLASE cDNA in an industrial baker's yeast strain.

The Aspergillus oryzae alpha-amylase cDNA was placed under the control of the Saccharomyces cerevisiae actin promoter (pACT1) and introduced into the ribosomal DNA locus of an industrial baker's yeast strain. To obtain a strain eligible for commercial use, we constructed an integrative cassette lacking bacterial DNA sequences but containing the alpha-amylase cDNA and ribosomal DNA sequences to target the integration to this locus. High-copy-number integrants were obtained including a defective TRP1d promoter in the integrative cassette. We selected one transformant, Rib-AMY (CECT10872), in which the multi-integrated sequences were stable even after 200 generations of growth in nonselective medium. This transformant also expressed and secreted high levels of alpha-amylase. Bread made with this strain had a higher volume, lower density, and softer crumbs than bread made with a control strain. The Rib-AMY transformant also was useful in retarding bread firming. This new strain fulfills all the requirements for commercial utilization and should reduce or eliminate the requirement for addition of exogenous alpha-amylase to the flour, reducing allergenic work-related symptoms due to this enzyme.

Aspergillus oryzae↗

PubMedDate not supplied

Determinants of wound infections after surgery for breast cancer.

BACKGROUND: There are evident risks factors stemming from any the local post-surgical infection, which merits analysis of morbidity derived from conservative and radical surgery after treatment of the operable breast cancer. STUDY DESIGN: 107 patients with breast cancer recruited between November 1995 and October 1997 were evaluated. From that total, 36 patients were subjected to quadrantectomy, axillary lymphadenectomy and postoperative radiation while 71 patients were subjected to modified radical mastectomy. RESULTS: 7 % percent of patients who underwent modified radical surgery succumbed to postoperative local infection vs. 17 % of patients subjected to breast conserving therapies. Local infections were associated with the older patients (63 vs. 54 years old, p=0.03) and greater obesity (p=0.04). Upon logistic regression analysis conservative surgery exhibited adjusted Odds Ratio of 6.42 (1.27-32.28), p =0.02. In postoperative infection group average hospital stay increased by 6 days (p= 0.0001). CONCLUSION: In the present study main risk factors associated with postsurgical wound infection after breast cancer treatment proved to be advanced age, greater obesity and conservative surgery. Nonetheless, more studies are necessary to confirm current findings.

Breast Neoplasms↗

PubMedDate not supplied

Do the conventional clinicopathologic parameters predict for response and survival in head and neck cancer patients undergoing neoadjuvant chemotherapy?

AIMS AND BACKGROUND: Neoadjuvant chemotherapy for head and neck carcinoma is still an important treatment modality. The prognostic value of patient and tumor parameters has been extensively evaluated in several trials, yielding mixed results. We report the prognostic factors emerging from a group of patients undergoing neoadjuvant chemotherapy. PATIENTS AND METHODS: From April 1986 to June 1992, 149 consecutive patients received cisplatin-5-fluorouracil-based neoadjuvant chemotherapy. After four courses of chemotherapy, patients underwent local-regional treatment with surgery, radiation or both. A variety of patient and tumor characteristics were evaluated as predictors for response to chemotherapy and survival. RESULTS: The complete response, partial response and no response rates to NAC were 52%, 33% and 15%, respectively. No parameters predicted response to chemotherapy. At a maximum follow-up of 87 months, overall survival was 39% and disease-free survival was 49%. Variables shown to be predictors of survival in univariate analyses were age, performance status, histology, site, T, N, stage, and response to chemotherapy. Using the Cox regression analysis, only complete response to induction chemotherapy (P = 0.0006), performance status (P = 0.03), stage (P = 0.01), age (P = 0.03) and primary tumor site (P = 0.04) emerged as independent prognostic factors for survival. CONCLUSIONS: Complete response to chemotherapy was confirmed as the strongest prognostic factor influencing survival. However, conventional clinicopathologic factors did not predict response, hence, potential prognostic biologic and molecular factors for response must be sought. At present, much effort must be made for the improvement of the complete response rate, which seems to be a requisite to prolong survival.

Adult↗

PubMedDate not supplied

Evolution of IgG antibody response against Toxoplasma gondii tissue cyst in acute and chronic human infections.

The recognition profile of the tissue cysts antigens by IgG antibodies was studied during acute and chronic human toxoplasmic infection. Thus the IgG response against Toxoplasma gondii was investigated by immunoblotting in two patients accidentally infected with the RH strain as well as in group of naturally infected patients at acute and chronic phase. There was an overall coincidence of molecular mass among antigens of tachyzoites and tissue cysts recognized by these sera, however, they appear not to be the same molecules. The response against tissue cysts starts early during acute infection, and the reactivity of antibodies is strong against a wide range of antigens. Six bands (between 82 and 151 kDa) were exclusively recognized by chronic phase sera but only the 132 kDa band was positive in more than 50% of the sera analysed. A mixture of these antigens could be used to discriminate between the two infection phases. The most important antigens recognized by the acute and the chronic phase sera were 4 clusters in the ranges 20-24 kDa, 34-39 kDa, 58-80 kDa and 105-130 kDa as well as two additional antigens of 18 and 29 kDa. Both accidentally infected patients and some of the naturally infected patients showed a weak specific response against tissue cyst antigens.

Acute Disease↗

PubMedDate not supplied

IgG recognizing 21-24 kDa and 30-33 kDa tachyzoite antigens show maximum avidity maturation during natural and accidental human toxoplasmosis.

We describe the avidity maturation of IgGs in human toxoplasmosis using sequential serum samples from accidental and natural infections. In accidental cases, avidity increased continuously throughout infection while naturally infected patients showed a different profile. Twenty-five percent of sera from chronic patients having specific IgM positive results could be appropriately classified using exclusively the avidity test data. To take advantage of the potentiality of this technique, antigens recognized by IgG showing steeper avidity maturation were identified using immunoblot with KSCN elution. Two clusters of antigens, in the ranges of 21-24 kDa and 30-33 kDa, were identified as the ones that fulfill the aforementioned avidity characteristics.

Acute Disease↗

PubMedDate not supplied

Characterization and optimization of bovine Echinococcus granulosus cyst fluid to be used in immunodiagnosis of hydatid disease by ELISA.

The aim of this work was to assess the influence in the diagnostic value for human hydatid disease of the composition of bovine hydatid cyst fluid (BHCF) obtained from fertile (FC) and non-fertile cysts (NFC). Eight batches from FC and 5 from NFC were prepared and analysed with respect to chemical composition: total protein, host-derived protein, carbohydrate and lipid contents. No differences were observed in the first two parameters but carbohydrate and lipid contents were shown to be higher in batches from FC than in those from NFC. Bands of 38 and 116 kD in SDS-PAGE profiles were observed to be present in BHCF from FC only. Two pools were prepared from BHCF batches obtained from FC (PFC) and NFC (PNFC), respectively. Antigen recognition patterns were analysed by immunoblot. Physicochemical conditions for adsorption of antigens to the polystyrene surface (ELISA plates) were optimized. The diagnostic value of both types of BHCF as well as the diagnostic relevance of oxidation of their carbohydrate moieties with periodate were assessed by ELISA using 42 serum samples from hydatid patients, 41 from patients with other disorders, and 15 from healthy donors. Reactivity of all sera against native antigen were tested with and without free phosphorylcholine. The best diagnostic efficiency was observed using BHCF from periodate-treated PFC using glycine buffer with strong ionic strength to coat ELISA plates.

Animals↗

PubMedDate not supplied

[What role does eosinophil cationic protein have in the evaluation of asthma severity?].

The revival of the concept of asthma as being a chronic inflammatory disease has led to the development of methods to monitor and control the underlying inflammatory process. The eosinophil is one of the main protagonists in this process, the control of the proteins that are released by the eosinophil when it is activated (more specifically the ECP) could provide valuable information for this purpose. However, the great complexity of the asthmatic inflammation obliges us to take into account a series of considerations, aiming at nor attributing nor expecting information from the ECP that it cannot provide. The ECP has been used in the diagnosis of asthma, in the prognosis of wheezing infants, as a marker for the exposure to the allergen, to evaluate the results of the provocation tests, with the clinical and functional seriousness of asthma, to control the effect of different anti-inflammatory treatments as well as to monitor the performance of these treatment, etc. However, it is necessary to take into account the fact that this marker is a reflection of the activation of the eosinophil, without specifying where the said activation has been produced and therefore there are studies that document increase of the ECP in relation to other conditions besides asthma (rhinitis, atopic dermatitis, the mother's addiction to tobacco, parasitosis, atopia, etc.). On the other hand there is a certain overlapping of the ECP values of patients with the active illness and with normal controls. Even patients with strong signs of asthma on occasions show normal ECP values, which could reflect that, in some cases, the eosinophilic inflammation may not be a relevant factor or, at least, it isn't a predominant one. On the other hand, when the asthma is being controlled, the determination of the ECP in induced sputum, could have a more precise diagnosis, while it could be a more exact reflection of the inflammation that exists on a local level, although this is a fact that needs to be proved subsequently. The principal usefulness of the ECP is for the individual follow up and control of each specific patient. Whatever the case, and given that the underlying asthmatic inflammation is a very complex process, in order to control it, a suitable combination of different markers is needed, which predictably improves the reliability of the diagnosis and prognosis and allows for a more precise control of the asthmatic inflammation.

Asthma↗