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Biomedical subjects

J Huguet

Publications and source records attributed to J Huguet.

At least 19 recordsLinked to original sources

Cystectomy in patients with high risk superficial bladder tumors who fail intravesical BCG therapy: pre-cystectomy prostate involvement as a prognostic factor.

PURPOSE: To review understaging and outcome of patients who underwent radical cystectomy (RC) for high risk superficial bladder cancer after bacillus Calmette-Guérin (BCG) failure. PATIENTS AND METHODS: We carried out a retrospective study of 62 cases in which RC was indicated for clinical stage Tis, Ta, T1 transitional cell bladder tumors that failed transurethral resection (TUR) and BCG treatment. We used BCG (81 mg/Connaught BCG) in patients with superficial grade 3 tumors and CIS. We considered BCG failure a high-grade recurrence at 3 months of the first BCG course or after 2 courses. RC indications, correlation between their clinical and pathological stage and the ensuing progress were analyzed. We assessed the existence of any pre-cystectomy clinical or pathological factor related to understaging and survival. RESULTS: RC was performed in 22 patients with carcinoma in situ (CIS) (35%), 7 with Ta (11,2%), 31 with T1 (50%), and 2 with Tx tumors (3%). All 62 but one were high-grade tumors (grade 3 and/or CIS). Tumor was clinically understaged with stages pT2 or greater on the RC specimen in 17 patients (27%). The presence of tumor in the prostatic urethra at the moment of endoscopic staging before RC was the only factor associated with clinical understaging (p=0.003) and shorter survival (p<0.0002). Five-year disease-specific survival rate was significantly lower in understaged (38%) as compared with not-understaged patients (90%) after a median follow-up of 40-months (range 1-142) (p=0.006). Overall five-year disease-specific survival was 79%. CONCLUSIONS: RC should be performed prior to progression in high risk superficial tumors that fail after TUR and BCG. In patients with clinical and pathological nonmuscle invasive disease, RC provides an excellent disease-free survival. One third of patients with HRSBT who underwent RC after BCG failure were understaged and had a shorter survival. Tumor in the prostatic urethra at endoscopic staging was the only factor associated to understaging and shorter survival.

Adjuvants, Immunologic↗

Genetic syndromes mimic congenital infections.

Genetic syndromes that mimic congenital infection are important to recognize because of the associated risk of recurrence. We describe two brothers born to consanguineous parents with clinical features suggestive of intrauterine infection but with negative serologic investigations. Our observations suggest that Aicardi-Goutieres syndrome (AGS) and pseudo-TORCH syndrome likely represent the same disorder.

Adult↗

[Living donor nephrectomy for kidney transplantation. Experience in the first two years].

OBJECTIVES: Laparoscopic surgery offers potential advantages in terms of diminishment of postoperative pain, shorter hospital stay, faster convalescence, and better cosmetic results. These advantages may increase kidney donation, making donation be accepted by more candidates. We report our first 2 years' experience with laparoscopic donor nephrectomy METHODS: Between March 2002 and February 2004 we performed 38 laparoscopic living donor nephrectomies for kidney transplantation. The technique of choice was the transperitoneal laparoscopic approach with four trocars, usually three of them from the start of the procedure--two 10-12 mm and one 5 mm--, and a 6.5 cm perumbilical midline incision for kidney retrieval at the end of the procedure. RESULTS: Receptor and donor survivals were 100%. Graft survival was 97.6%. There was not any case of delayed graft function. Donor: Mean operative time was 161 minutes (115-260). Mean estimated blood loss was 270 ml (100-1200). Three patients required blood transfusions, 2 units of packed red blood cells each. Mean hospital stay was 5.1 days (3-11). Mean warm ischemia time was 3.2 min. (2-10). Conversion to open surgery was necessary in four cases. Receptor: there have been three significant complications requiring surgical repair: one case of low arterial flow, one vesico ureteral leak, and one midurethra stenosis. Initial renal function: mean serum creatinine at one month was 147mmol/l, with a trend to improve to 126 mmol/l at one year, which is considered optimum. First postoperative day mean serum creatinine was 192mmol/l and the nadir was on second postoperative day with a value of 152mmol/l. CONCLUSIONS: We believe laparoscopic living donor nephrectomy is a real alternative to open surgery because it offers better recovery to the donor with the same capacity to preserve renal function in the receptor.

Humans↗

Endoscopic approach of the overinfected urachal cyst.

The overinfected urachus cyst presents itself in quite different forms. In spite of the accuracy of ultrasound and abdominal CT scan, sometimes the diagnosis of this urachal abnormality is imprecise and even misleading. This article describes the transvesical endoscopic drainage of an infected urachal cyst with optimum result upon the treatment of this condition.

Adult↗

Modulation of lipase properties in macro-aqueous systems by controlled enzyme immobilization: enantioselective hydrolysis of a chiral ester by immobilized Pseudomonas lipase.

Lipase from Pseudomonas fluorescens (PFL) has been immobilized by using different immobilization protocols. The catalytic behavior of the different PFL derivatives in the hydrolytic resolution of fully soluble (R,S) 2-hydroxy 4-phenyl butanoic acid ethyl ester (HPBE) in aqueous medium was analyzed. The soluble enzyme showed a significant but low enantioselectivity, hydrolyzing the S isomer more rapidly than the R-isomer (E = 7). The enzyme, immobilized via a limited attachment to a long and flexible spacer arm, showed almost identical activity and specificity to the soluble enzyme. However, other derivatives, e.g. PFL adsorbed on supports covered by hydrophobic moieties (octyl, decaoctyl), exhibited significant hyperactivation on immobilization (approximately 7-fold). Simultaneously, the enantioselectivity of the PFL-immobilized enzyme was significantly improved (from E = 7 to E = 80). By using such derivatives, almost pure R ester isomer (e.e. > 99%) has been obtained after 55% hydrolysis of the racemic mixture of a solution of 10% (w/v) (R,S) HPBE. The derivatives could be used for 10 cycles without any significant decrease in the activity of the biocatalyst.

Journal Article↗

Determination of substituents distribution in carboxymethylpullulans by NMR spectroscopy.

The distribution of carboxymethyl substituents in the alpha-(1 --> 6)-linked maltotriosyl repeating units of a carboxymethylpullulan (CMP) series was investigated by high resolution NMR spectroscopy on very short oligomers (DPn = 1.2-1.5) obtained by acid hydrolysis. A series of 2D NMR experiments on parent pullulan, hydrolysed pullulan and CMP was used to assign the proton and carbon chemical shifts of CMP acid hydrolysates. The degree of substitution (DS) and the relative distribution of -CH2COONa groups at OH-2, OH-3, OH-4 and OH-6 of glucose residues (DSi) were determined from 1H NMR measurements. From a set of CMP samples, widely different in degree of substitution, it was observed that the substitution at C-2 is predominant and decreases according to the order C-2 > C-3 > C-6 > C-4. Taking into account the availability of each OH group in the parent pullulan, an order of relative reactivity of hydroxyl groups is defined according to the relation: Ri = DSi/ni, where ni is the number of free OH groups in a maltotriose unit (MTU) for a given site C-i, the reactivity order was found to be OH-2 > OH-4 > OH-6 > OH-3.

Carbohydrate Sequence↗

A single step purification, immobilization, and hyperactivation of lipases via interfacial adsorption on strongly hydrophobic supports

A number of bacterial lipases can be immobilized in a rapid and strong fashion on octyl-agarose gels (e.g., lipases from Candida antarctica, Pseudomonas fluorescens, Rhizomucor miehei, Humicola lanuginosa, Mucor javanicus, and Rhizopus niveus). Adsorption rates in absence of ammonium sulfate are higher than in its presence, opposite to the observation for typical hydrophobic adsorption of proteins. At 10 mM phosphate, adsorption of lipases is fairly selective allowing enzyme purification associated with their reversible immobilization. Interestingly, these immobilized lipase molecules show a dramatic hyperactivation. For example, lipases from R. niveus, M. miehei, and H. lanuginosa were 6-, 7-, and 20-fold more active than the corresponding soluble enzymes when catalyzing the hydrolysis of a fully soluble substrate (0.4 mM p-nitrophenyl propionate). Even higher hyperactivations and interesting changes in stereospecificity were also observed for the hydrolysis of larger soluble chiral esters (e.g. (R,S)-2-hydroxy-4-phenylbutanoic ethyl ester). These results suggest that lipases recognize these "well-defined" hydrophobic supports as solid interfaces and they become adsorbed through the external areas of the large hydrophobic active centers of their "open and hyperactivated structure". This selective interfacial adsorption of lipases becomes a very promising immobilization method with general application for most lipases. Through this method, we are able to combine, via a single and easily performed adsorption step, the purification, the strong immobilization, and a dramatic hyperactivation of lipases acting in the absence of additional interfaces, (e.g., in aqueous medium with soluble substrate). Copyright 1998 John Wiley & Sons, Inc.

Journal Article↗

Human immunodeficiency virus type 1--serodiscordant couples can bear healthy children after undergoing intrauterine insemination.

OBJECTIVE: To use semen from men who were seropositive for human immunodeficiency virus type 1 (HIV-1) to inseminate their partners without infecting them. DESIGN: Prospective study. SETTING: Private practice. PATIENT(S): Sixty-three HIV-1-seropositive men and their HIV-1-seronegative female partners. INTERVENTION(S): The men provided 107 semen samples that were prepared with the use of the Percoll and swim-up techniques. The presence of HIV-1 was determined in the fraction of motile spermatozoa obtained after washing. If HIV-1 was not detected. IUI was performed in stimulated cycles. MAIN OUTCOME MEASURE(S): Human immunodeficiency virus type 1 RNA and DNA were detected with the use of the polymerase chain reaction technique modified for spermatozoa. RESULT(S): One hundred seven semen samples were washed. Human immunodeficiency virus type 1 was not detected in 101 samples (94.4%) and was detected in 6 samples (5.6%). In the latter cases, IUI was not performed. One hundred one IUI procedures were performed in 63 women. Thirty-one pregnancies resulted, for a pregnancy rate of 30.7% per cycle and 49.2% per inseminated woman. Thirty-seven healthy children were born. The results of tests for the detection of HIV-1 and antibodies to HIV-1 in the inseminated women were negative. CONCLUSION(S): On the basis of these results, testing for HIV-1 with the use of the polymerase chain reaction technique on the semen fraction obtained after washing appears to prevent infection in the inseminated woman. This method makes it possible to help HIV-1-seropositive men to have children without infecting their female partners.

Adult↗

Pregnancy following intracytoplasmic sperm injection from an HIV-1-seropositive man.

The first pregnancy achieved in a seronegative woman following in-vitro fecundation through intracytoplasmic sperm (ICSI) injection from a man with autoimmune deficiency syndrome (AIDS; HIV-1 carrier) is reported. The semen was prepared by PureSperm and swim-up techniques. Some of the motile spermatozoa obtained were used to detect the presence of HIV-1 using the polymerase chain reaction technique. HIV-1 in DNA or RNA form was not detected using this technique. The remaining spermatozoa were frozen. Ovarian stimulation in the woman was performed with long-protocol analogues and gonadotrophins. Thirteen mature oocytes were recovered, into which the thawed spermatozoa were microinjected. Nine embryos were obtained. Four were frozen, four transferred and one discarded. The woman became pregnant. Analyses for HIV-1 in the woman, performed in the first and third months of pregnancy, gave negative results. This case provides further experience with washed semen of sufficient quality for performing artificial insemination in HIV-1-serodiscordant couples (101 inseminations, 31 pregnancies, 28 deliveries, 37 babies, all healthy). In women with obstructed Fallopian tubes, or when the semen is not of sufficient quality for artificial insemination techniques to be performed, ICSI can be carried out using frozen, HIV-1-free semen.

Acquired Immunodeficiency Syndrome↗

Immunological quantitation of nuclear steroid receptors to optimize the biological classification of breast tumors.

We used immunological methods to determine cytosolic and nuclear steroid receptors to evaluate the advantages of nuclear receptor measurement in the selection of breast cancer patients for treatment. Around 75% of tumors showed coincidence between nuclear and cytosolic receptors (+/+ or -/-) for estrogen receptor (ER) and for progesterone receptor (PgR). Only cytosolic receptors were detected in around 20% of tumors. Distributed in the ER/PgR phenotypes according to the nuclear or cytosolic receptors, 64% of tumors remained in the same subgroup, whereas 16% of tumors were classified as hormone dependent according to cytosolic and independent according to nuclear receptors, which could be considered as 'false-positive' results. 6% of tumors would be classified as negative according to cytosolic receptors but positive according to nuclear receptors and would correspond to 'false-negative' results by conventional methods. Cytosolic receptor results may overrate the hormone dependence and cause some 'misclassifications' of patients. This could partially explain the lack of response to therapy in some cases.

Adult↗

Associative behaviour of hydrophobically modified carboxymethylpullulan derivatives.

Hydrophobically modified carboxymethylpullulan (HMCMP) samples were obtained by reaction of small amounts of C16 alkylamine on carboxylic groups of the corresponding polyacid. The molar contents of alkyl chains ranged from 1.3 to 6.8% with respect to the anhydroglucose units (AGU) and the degree of substitution (DS) of carboxylic groups varied from 0.76 to 0.84. Solution properties of the sodium salt of HMCMPs were studied mainly by viscometric and size-exclusion chromatography/light-scattering methods. The low-shear viscosity of modified pullulans in 0.1 M NaCl solutions drastically increases with the hydrophobic content and polymer concentrations and the 6.8% modified sample has a quite pseudoplastic behaviour. These data showed that the polymers aggregated intermolecularly and displayed a compact globular structure in dilute solution. Furthermore, addition of NaCl or ethanol induced a decrease in viscosity although the molecular weights remained approximately constant. These results are consistent with a collapse of the polysaccharide aggregates.

Amines↗

Expression of ecto-adenosine deaminase and CD26 in human T cells triggered by the TCR-CD3 complex. Possible role of adenosine deaminase as costimulatory molecule.

The expression of surface adenosine deaminase (ADA) and CD26 in activated human T cells was studied by flow cytometry. PBLs and CD3+ or CD4+ cells, when subjected to a variety of stimuli (anti-CD3 Abs plus IL-2 or phorbol esters), presented two structurally different cell populations, which differed in size and cellular complexity (populations B1 and B2). In PBLs triggered by an anti-CD3 mAb there was no significant increase of expression of either surface ADA or CD26 in cells of population B1, whose structure is similar to that of nonstimulated cells. In contrast, there was a significant increase in the percentage of expression of ADA and CD26 in the population B2, which corresponds to structurally more complex and larger cells. In the case of activation via TCR-CD3 but in the presence of IL-2 or via phorbol esters, the increase was found in cells from both populations, but B2 cells always showed a higher percentage of expression than B1 cells. The results of increased expression of surface ADA and CD26 were similar in whole T cells or in purer preparations such as CD3+ or CD4+ lymphocytes. Polyclonal Abs against ADA were not able to induce an activation response in T cells even when cross-linked by a secondary Ab. Interestingly, these Abs produced anergy in CD4+ cells subjected to an anti-CD3 stimulus. In contrast, addition of ADA produced an enzyme-independent synergism in the response through the TCR-CD3 complex. In T cells, ADA and CD26 colocalized on the surface of T cells; thus, the effect of exogenous ADA seems to be mediated by CD26 molecules that are not interacting with endogenous ADA (spare CD26 molecules). The presence of spare CD26 molecules on the surface of CD4+ cells was demonstrated by flow cytometry in the presence of exogenous ADA and also by confocal microscopy. The set of results strongly indicates that ADA binding to CD26 produces a costimulatory response in T cell activation events.

Adenosine Deaminase↗

Surface adenosine deaminase. A novel B-cell marker in chronic lymphocytic leukemia.

Previous studies found that ADA is present on the surface of mononuclear blood cells from healthy patients. Because the expression of this surface antigen depends upon the cell type, the presence of ADA on the plasma membrane of cells from patients with malignant hematologic diseases was studied by flow cytometry. The highest percentage of expression was found in CLL, whereas the lowest was found in T-cell-derived malignancies. The enzyme expression in immortalized cell lines showed a similar pattern, with the highest expression (95% +/- 5%) in the SKW64 B-derived cell line, the lowest (15% +/- 5%) in Jurkat T-lymphoma derived cells, and the intermediate (32% +/- 8%) in K562 cells derived from a chronic myelogenous leukemia. Double labeling ADA/CD5 and ADA/CD19, as well as the correlation of ADA expression with the expression of other surface markers, indicate that surface ADA might be considered a novel marker for CLL.

Adenosine Deaminase↗

[Open surgery of calculus of the kidney pelvis. Results and complications in the era of extracorporeal lithotripsy].

From 1989 to 1993, 11,535 extracorporeal shock wave lithotripsy (ESWL) sessions were performed in 6,066 patients in our centre. In contrast, simple or extended pyelolithotomy was the treatment of choice in a small group of patients. The object of this paper is to review the results and complications of this type of operation. A total of 57 pyelolithotomies was performed, including 33 extended pyelolithotomies, in 55 patients (26 men and 29 women). The mean age of the series was 50 years (range: 5-77). The operated kidney was the left kidney in 34 cases and the right kidney in 23 cases. The mean interval between the diagnosis and the operation was 31 months. 64% of the patients had history of stone disease (33 ipsilateral and 2 bilateral). Thirteen patients had already been treated by ESWL (10 patients), percutaneous nephrolithotomy (PNL) (2 cases) and one case had previously undergone pyelolithotomy. A urinary tract infection was observed in 26 patients (47%). The incision consisted of lumbotomy in 40 patients (71%), or a subcostal incision in 12 cases. The mean operating time was 161 minutes (range: 90-245) and the mean intraoperative bleeding was 500 ml. Thirteen patients were transfused (23%). The renal artery had to be clamped in 9 cases (for an average of 19 minutes). After a mean follow-up of 20 month, 50.8% of patients were stone-free. 49% of patients presented residual caliceal fragments on the immediate postoperative X-ray. One patient had to be treated by percutaneous nephrolithotomy and ESWL. 12 cases (41% of patients with residual fragments) required subsequent treatment by ESWL and the remaining patients with residual fragments were asymptomatic and did not receive any further treatment. The immediate postoperative complication rate was 17% and 10% of patients developed late postoperative complications. Simple or extended pyelolithotomy remains an effective treatment for stones of the renal pelvis, either on its own or associated with ESWL. However, the possibility of serious complications demands meticulous patient selection.

Adolescent↗

Surface expression of adenosine deaminase in mitogen-stimulated lymphocytes.

Adenosine deaminase (ADA) expression on the surface of mitogen-stimulated lymphocytes was studied by flow cytometry. The gate for lymphocytes was located by cell size (forward scatter), cytoplasmic complexity (side scatter) and by expression of the markers CD2, CD4, CD8 and CD19. After mitogenic proliferation two populations appeared, one corresponding to non-stimulated cells, and the other consisting of larger cells which showed relatively high expression of adenosine deaminase on their surface. The increase was similar to that observed for CD71 expression, and paralleled the increase in 3H-thymidine incorporation. There was a correlation between ADA and CD71 expression (r = 0.92 for phytohaemagglutinin (PHA) and 0.97 for pokeweed mitogen (PWM)). These results suggest a role for ecto-adenosine deaminase in lymphocyte proliferation and/or triggering.

5'-Nucleotidase↗