Biomedical subjects

O Alvarez

Publications and source records attributed to O Alvarez.

At least 19 recordsLinked to original sources

PubMed2003

Lectin typing of five medically important Candida species.

This study reports the patterns of agglutination of 93 clinical Candida isolates by 14 commercial lectins. The isolates were of the species Candida albicans (55), C. tropicalis (12), C. guilliermondii (10). C. glabrata (eight) and C. parapsilosis (eight). Hundred percent of isolates were agglutinated, at least, by a panel of three lectins: Canavalia ensiformis (ConA), Lens culinaris (LCA) and Pisum sativum (PSA), all of them with alpha-D-mannose specificity. In addition, another panel of three lectins could distinguish between C. glabrata, C. tropicalis and C. albicans. Lectin typing may be of potential value for taxonomic and epidemiological studies of yeasts in clinical laboratories.

Agglutination↗

PubMed2001

Periodic perturbations in Shaker K+ channel gating kinetics by deletions in the S3-S4 linker.

Upon depolarization positive charges contained in the transmembrane segment S4 of voltage-dependent channels are displaced from the cytoplasmic to the external milieu. This charge movement leads to channel opening. In Shaker K+ channels four positively charged arginines in the S4 domain are transferred from the internal to the external side of the channel during activation. The distance traveled by the S4 segment during activation is unknown, but large movements should be constrained by the S3-S4 linker. Constructing deletion mutants, we show that the activation time constant and the midpoint of the voltage activation curve of the Shaker K+ channel macroscopic currents becomes a periodic function of the S3-S4 linker length for linkers shorter than 7 aa residues. The periodicity is that typical of alpha-helices. Moreover, a linker containing only 3 aa is enough to recover the wild-type phenotype. The deletion method revealed the importance of the S3-S4 linker in determining the channel gating kinetics and indicated that the alpha-helical nature of S4 extends toward its N terminus. These results support the notion that a small displacement of the S4 segment suffices to displace the four gating charges involved in channel opening.

Animals↗

PubMed2001

Initial experience with Song's covered duodenal stent in the treatment of malignant gastroduodenal obstruction.

PURPOSE: Initial experience with use of Song's covered duodenal stent in the treatment of malignant gastroduodenal obstruction is reported. MATERIALS AND METHODS: Sixteen consecutive patients with malignant gastroduodenal obstruction were treated with peroral placement of Song's covered duodenal stent. The mean age was 58 years (range, 28-90 y). Gastroduodenal obstruction was caused by gastric (n = 8), metastatic (n = 2), gallbladder (n = 3), pancreatic (n = 2), or ampullary (n = 1) cancer. The disease was considered inoperable in all patients. With use of a flexible 20-F introducing system, seven fully covered, three uncovered, and 10 partially covered duodenal stents were placed under fluoroscopic guidance. RESULTS: The technical success rate was 94% (15 of 16) with no major complications. Symptoms of gastroduodenal obstruction improved in 14 patients. Stent migration was observed in three of seven fully covered stents. Patients with migrated stents required endoscopic stent removal and placement of uncovered duodenal stents. Tumor ingrowth was observed in two thirds of uncovered stents. In the 10 procedures with partially covered duodenal stents, no migration or tumor ingrowth was observed. All patients died 1-48 weeks (mean, 12 weeks) after stent placement. CONCLUSION: Peroral placement of Song's covered duodenal stent is a feasible and effective method of palliation in the majority of patients with malignant gastroduodenal obstruction. Migration of fully covered stents and tumor ingrowth of uncovered stents are important limitations that can be overcome with the use of a partially covered duodenal stent.

Adult↗

PubMed2001

Traumatic disruption of the pancreatic duct: diagnosis with MR pancreatography.

OBJECTIVE. We evaluated the ability of MR pancreatography to reveal traumatic disruptions of the pancreatic duct compared with retrograde pancreatography. CONCLUSION. MR pancreatography is an adequate noninvasive test for the detection of complete traumatic disruptions of the main pancreatic duct. MR pancreatography is especially useful for delineating the segments of the duct that cannot be evaluated with retrograde pancreatography.

Adolescent↗

PubMed2000

Modulation of the Shaker K(+) channel gating kinetics by the S3-S4 linker.

In Shaker K(+) channels depolarization displaces outwardly the positively charged residues of the S4 segment. The amount of this displacement is unknown, but large movements of the S4 segment should be constrained by the length and flexibility of the S3-S4 linker. To investigate the role of the S3-S4 linker in the ShakerH4Delta(6-46) (ShakerDelta) K(+) channel activation, we constructed S3-S4 linker deletion mutants. Using macropatches of Xenopus oocytes, we tested three constructs: a deletion mutant with no linker (0 aa linker), a mutant containing a linker 5 amino acids in length, and a 10 amino acid linker mutant. Each of the three mutants tested yielded robust K(+) currents. The half-activation voltage was shifted to the right along the voltage axis, and the shift was +45 mV in the case of the 0 aa linker channel. In the 0 aa linker, mutant deactivation kinetics were sixfold slower than in ShakerDelta. The apparent number of gating charges was 12.6+/-0.6 e(o) in ShakerDelta, 12.7+/-0.5 in 10 aa linker, and 12.3+/-0.9 in 5 aa linker channels, but it was only 5.6+/-0.3 e(o) in the 0 aa linker mutant channel. The maximum probability of opening (P(o)(max)) as measured using noise analysis was not altered by the linker deletions. Activation kinetics were most affected by linker deletions; at 0 mV, the 5 and 0 aa linker channels' activation time constants were 89x and 45x slower than that of the ShakerDelta K(+) channel, respectively. The initial lag of ionic currents when the prepulse was varied from -130 to -60 mV was 0.5, 14, and 2 ms for the 10, 5, and 0 aa linker mutant channels, respectively. These results suggest that: (a) the S4 segment moves only a short distance during activation since an S3-S4 linker consisting of only 5 amino acid residues allows for the total charge displacement to occur, and (b) the length of the S3-S4 linker plays an important role in setting ShakerDelta channel activation and deactivation kinetics.

Amino Acids↗

PubMed2000

Detection of choledocholithiasis with MR cholangiography: comparison of three-dimensional fast spin-echo and single- and multisection half-Fourier rapid acquisition with relaxation enhancement sequences.

PURPOSE: To compare the performance of three pulse sequences commonly used at magnetic resonance (MR) cholangiography in the diagnosis of choledocholithiasis. MATERIALS AND METHODS: MR cholangiography was performed in 57 patients who were suspected of having choledocholithiasis and referred for endoscopic retrograde cholangiography. Non-breath-hold three-dimensional fast spin-echo, breath-hold single-section half-Fourier rapid acquisition with relaxation enhancement (RARE), and breath-hold multisection half-Fourier RARE sequences were compared. Two radiologists independently interpreted the MR cholangiograms. Evaluated diagnostic performance parameters included sensitivity, specificity, receiver operating characteristic (ROC) curves, and interobserver agreement (kappa statistics). Endoscopic retrograde cholangiography was the standard of reference. RESULTS: Eight patients were excluded because of incomplete MR examinations (n = 4) or failure in the cannulation of the bile duct at retrograde cholangiography (n = 4). In 49 patients, the three MR cholangiographic sequences were completed successfully. In 24 (49%) of these patients, retrograde cholangiography demonstrated stones. Sensitivity and specificity of MR cholangiography exceeded 90%, and the area under the ROC curve was greater than 0.95 for both radiologists and for the three sequences. Interobserver agreement for presence of bile duct stones was at least 0.80 (very good) for the three sequences. CONCLUSION: The three MR cholangiographic sequences had similarly high sensitivities and specificities for the detection of choledocholithiasis.

Adolescent↗

PubMed2000

Diagnosing bile duct stones: comparison of unenhanced helical CT, oral contrast-enhanced CT cholangiography, and MR cholangiography.

OBJECTIVE: In this investigation we compared the diagnostic performance of unenhanced helical CT, oral contrast-enhanced CT cholangiography, and MR cholangiography for the diagnosis of choledocholithiasis. SUBJECTS AND METHODS: Fifty-one patients referred for endoscopic retrograde cholangiography of suspected biliary stones were studied with unenhanced helical CT, MR cholangiography, and helical CT performed after oral administration of a cholangiographic contrast agent (iopodic acid). The studies were randomized for interpretation. Two radiologists evaluated the images by consensus and determined the presence and location of stones. We used retrograde cholangiography findings as the standard of reference. Sensitivity and specificity (with 95% confidence intervals [CIs]) of the three examinations were calculated and compared using the exact form of the McNemar test. RESULTS: Bile duct stones were revealed with retrograde cholangiography in 26 patients (51%). Sensitivity was 65% (95% CI, 44.4-82%) for unenhanced helical CT, 92% (95% CI, 73-99%) for CT cholangiography, and 96% (95% CI, 78-99%) for MR cholangiography. Specificity was 84% (95% CI, 63-95%) for unenhanced helical CT, 92% (95% CI, 73-99%) for CT cholangiography, and 100% (95% CI, 83-100%) for MR cholangiography. The sensitivity of CT cholangiography and MR cholangiography was significantly higher than that of unenhanced helical CT (p<0.01). Differences in specificity were not significant. CONCLUSION: Our results indicate that oral contrast-enhanced CT cholangiography and MR cholangiography are significantly more sensitive than unenhanced helical CT for the detection of bile duct calculi.

Adolescent↗

PubMed1999

Localization of the K+ lock-In and the Ba2+ binding sites in a voltage-gated calcium-modulated channel. Implications for survival of K+ permeability.

Using Ba2+ as a probe, we performed a detailed characterization of an external K+ binding site located in the pore of a large conductance Ca2+-activated K+ (BKCa) channel from skeletal muscle incorporated into planar lipid bilayers. Internal Ba2+ blocks BKCa channels and decreasing external K+ using a K+ chelator, (+)-18-Crown-6-tetracarboxylic acid, dramatically reduces the duration of the Ba2+-blocked events. Average Ba2+ dwell time changes from 10 s at 10 mM external K+ to 100 ms in the limit of very low [K+]. Using a model where external K+ binds to a site hindering the exit of Ba2+ toward the external side (Neyton, J., and C. Miller. 1988. J. Gen. Physiol. 92:549-568), we calculated a dissociation constant of 2.7 mircoM for K) at this lock-in site. We also found that BK(Ca) channels enter into a long-lasting nonconductive state when the external [K+] is reduced below 4 microM using the crown ether. Channel activity can be recovered by adding K+, Rb+, Cs+, or NH4+ to the external solution. These results suggest that the BK(Ca) channel stability in solutions of very low [K+] is due to K+ binding to a site having a very high affinity. Occupancy of this site by K+ avoids the channel conductance collapse and the exit of Ba2+ toward the external side. External tetraethylammonium also reduced the Ba2+ off rate and impeded the channel from entering into the long-lasting nonconductive state. This effect requires the presence of external K+. It is explained in terms of a model in which the conduction pore contains Ba2+, K+, and tetraethylammonium simultaneously, with the K+ binding site located internal to the tetraethylammonium site. Altogether, these results and the known potassium channel structure (Doyle, D.A., J.M. Cabral, R.A. Pfuetzner, A. Kuo, J.M. Gulbis, S.L. Cohen, B.T. Chait, and R. MacKinnon. 1998. Science. 280:69-77) imply that the lock-in site and the Ba2+ sites are the external and internal ion sites of the selectivity filter, respectively.

Algorithms↗

PubMed1999

Lectin typing of methicillin-resistant Staphylococcus aureus.

This study reports the patterns of agglutination of 77 clinical isolates of methicillin-resistant Staphylococcus aureus by 32 commercially available lectins. Cell suspensions were not pre-treated. Each isolate was cultured on three media: Columbia blood agar, trypticase-soy agar and Chapman Stone agar. The lectins agglutinating each isolate varied widely depending on culture medium; only five isolates were agglutinated by the same set of lectins regardless of the culture medium used. Lectin typing could be a useful epidemiological tool, but it is necessary to standardise assay conditions (notably culture medium) to enable meaningful comparison of the results produced by different research groups or centres.

Agglutination Tests↗

PubMed1998

Role of the S4 segment in a voltage-dependent calcium-sensitive potassium (hSlo) channel.

We investigated the role of individual charged residues of the S4 region of a MaxiK channel (hSlo) in channel gating. We measured macroscopic currents induced by wild type (WT) and point mutants of hSlo in inside-out membrane patches of Xenopus laevis oocytes. Of all the residues tested, only neutralizations of Arg-210 and Arg-213 were associated with a reduction in the number of gating charges as determined using the limiting slope method. Channel activation in WT and mutant channels was interpreted using an allosteric model. Mutations R207Q, R207E, and R210N facilitated channel opening in the absence of Ca2+; however, this facilitation was not observed in the channels Ca2+-bound state. Mutation R213Q behaved similarly to the WT channel in the absence of Ca2+, but Ca2+ was unable to stabilize the open state to the same extent as it does in the WT. Mutations R207Q, R207E, R210N, and R213Q reduced the coupling between Ca2+ binding and channel opening when compared with the WT. Mutations L204R, L204H, Q216R, E219Q, and E219K in the S4 domain showed a similar phenotype to the WT channel. We conclude that the S4 region in the hSlo channel is part of the voltage sensor and that only two charged amino acid residues in this region (Arg-210 and Arg-213) contribute to the gating valence of the channel.

Amino Acid Sequence↗

PubMed1998

Pore accessibility during C-type inactivation in Shaker K+ channels.

Shaker K+ channels inactivate through two distinct molecular mechanisms: N-type, which involves the N-terminal domain and C-type that appears to involve structural modifications at the external mouth of the channel. We have tested pore accessibility of the Shaker K+ channel during C-type inactivation using Ba2+ as a probe. We determined that external Ba2+ binds to C-type inactivated channels forming an extremely stable complex; i.e. there is Ba2+ trapping by C-type inactivated channels. The structural changes Shaker channels undergo during C-type inactivation create high energy barriers that hinder Ba2+ exit to either the extracellular solution or to the intracellular solution.

Barium↗

PubMed1998

Rapid healing of venous ulcers and lack of clinical rejection with an allogeneic cultured human skin equivalent. Human Skin Equivalent Investigators Group.

OBJECTIVE: To test the safety, efficacy, and immunological impact of a cultured allogeneic human skin equivalent (HSE) in the treatment of venous ulcers. DESIGN: Prospective, randomized study. SETTING: Multicenter study in the outpatient setting. INTERVENTION: Each patient with a venous ulcer received either compression therapy alone or compression therapy and treatment with HSE. The patients were evaluated for HSE safety, complete (100%) ulcer healing, time to wound closure, wound recurrence, and immune response to the HSE. OUTCOME: The study was completed as planned in 293 randomized patients. RESULTS: Treatment with HSE was more effective than compression therapy in the percentage of patients healed by 6 months (63% vs 49%; P=.02, Fisher exact test, 2-tailed) and the median time to complete wound closure (61 days vs 181 days; P=.003, log-rank test). Treatment with HSE was superior to compression therapy in healing larger (> 1000 mm2; P=.02) and deeper ulcers (P=.003) and ulcers of more than 6 months' duration (P=.001). Occurrence of adverse events was similar in both groups. No symptoms or signs of rejection occurred in response to treatment with HSE, and no HSE-specific immune responses were detected in vitro to bovine collagen or to alloantigens expressed on keratinocytes or fibroblasts. CONCLUSIONS: Treatment with HSE healed venous ulcers more rapidly and in more patients than compression therapy alone. There was no clinical or laboratory evidence of rejection or sensitization in response to HSE application. These data suggest that HSE represents a significant advance in the treatment of venous ulcers, particularly those that are difficult to heal.

Aged↗

PubMed1997

Cation permeability of a cloned rat epithelial amiloride-sensitive Na+ channel.

1. Conductance of heterotrimeric rat epithelial Na+ channels (alpha, beta, gamma-rENaCs) for Li+ and Na+ in planar lipid bilayers was a non-linear function of ion concentration, with a maximum of 30.4 +/- 2.9 pS and 18.5 +/- 1.9 pS at 1 M Li+ and Na+, respectively. 2. The alpha, beta, gamma-rENaC conductance measured in symmetrical mixtures of Na(+)-Li+ (1 M) exhibited an anomalous mole fraction dependence, with a minimum at 4:1 Li+ to Na+ molar ratio. 3. Permeability ratios PK/PNa and PLi/PNa of the channel calculated from the bionic reversal potentials were dependent on ion concentration: PK/PNa was 0.11 +/- 0.01, and PLi/PNa was 1.6 +/- 0.3 at 50 mM; PK/PNa was 0.04 +/- 0.01 and PLi/PNa was 2.5 +/- 0.4 at 3 M, but differed from the ratios of single-channel conductances in symmetrical Li+, Na+ or K+ solutions. The permeability sequence determined by either method was Li+ > Na+ > K+ >> Rb+ Cs+. 4. Predictions of a model featuring two binding sites and three energy barriers (2S3B), and allowing double occupancy, developed on the basis of single ion current-voltage relationships, are in agreement with the observed conductance maximum in single ion experiments, conductance minimum in the mole fraction experiments, non-linearity of the current-voltage curves in bionic experiments, and the concentration dependence of permeability ratios. 5. Computer simulations using the 2S3B model recreate the ion concentration dependencies of single-channel conductance observed for the immunopurified bovine renal amiloride-sensitive Na+ channel, and short-circuit current in frog skin, thus supporting the hypothesis that ENaCs form a core conduction unit of epithelial Na+ channels.

Animals↗

PubMed1997

Proton therapy for pediatric cranial tumors: preliminary report on treatment and disease-related morbidities.

PURPOSE: Accelerated protons were used in an attempt to limit treatment-related morbidity in children with tumors in or near the developing brain, by reducing the integral dose to adjacent normal tissues. METHODS AND MATERIALS: Children treated with protons at Loma Linda University Medical Center between August 1991 and December 1994 were analyzed retrospectively. Twenty-eight children, aged 1 to 18 years, were identified as at risk for brain injury from treatment. Medical records, physical examinations, and correspondence with patients, their parents, and referring physicians were analyzed. The investigators tabulated post-treatment changes in pre-treatment signs and symptoms and made judgments as to whether improvement, no change, or worsening related to disease or treatment had supervened. Magnetic resonance images were correlated with clinical findings and radiographic impressions were tabulated. RESULTS: Follow-up ranged from 7 to 49 months (median 25 months). Four instances of treatment-related morbidity were identified. Forty-one instances of site-specific, disease-related morbidity were identified: 15 improved or resolved and 26 remained unchanged after treatment. Four patients had radiographic evidence of local failure. Three of these patients, including two with high-grade glioma, have died. CONCLUSION: Early treatment-related morbidity associated with proton therapy is low. Tumor progression remains a problem when treating certain histologies such as high-grade glioma. Escalating the dose delivered to target volumes may benefit children with tumors associated with poor rates of local control. Long-term follow-up, including neurocognitive testing, is in progress to assess integral-dose effects on cognitive, behavioral and developmental outcomes in children with cranial tumors.

Adolescent↗

PubMed1996

Cultured skin as a 'smart material' for healing wounds: experience in venous ulcers.

The healing of chronic wounds is a difficult and varied problem. The engineering of a cultured skin tissue offers an adaptive therapy for chronic wounds. Our hypothesis has been that living tissue can act as a 'smart material' to heal wounds. We have examined the healing characteristics of a bilayered cultured skin equivalent (Graftskin) in a controlled study and present clinical data from interim analyses for 233 patients over 6 months of treatment. All venous ulcer patients will be followed for up to 1 year. We report on three basic scenarios of healing: (i) promotion of healing by secondary intention, (ii) persistent biological wound closure with stimulation of underlying healing, and (iii) healing by frank graft take of the cultured material with remodelling of the tissue over time. Our results indicate that the cultured skin equivalent is responsive to individual wound conditions and thus acts as a 'smart material' in the chronic wound.

Adult↗

PubMed1995

Simultaneous recording of analog signals, audio and video using a VCR.

An interface to record analog signals from DC to 20 kHz on the stereo sound channels of video cassette recorders is described. This interface is an effective solution for the implementation of experiments requiring simultaneous recording of electrophysiological analog data, audio and video signals, since all these data are recorded on the same tape. The system requires the construction of a simple analog interface and minor modifications internal to the VCR, required to bypass non-linear elements present on the audio signal path. The particular solution used to adapt for Sony VCR model SLV-575UC is given in full detail. The general requirements the other VCR makes and models must fulfil to make the adaptation feasible and guidelines for the design of interfaces are discussed.

Analog-Digital Conversion↗

PubMed1995

Randomized double-blind crossover ondansetron-dexamethasone versus ondansetron-placebo study for the treatment of chemotherapy-induced nausea and vomiting in pediatric patients with malignancies.

PURPOSE: To determine whether the addition of dexamethasone to ondansetron (OND + DEX) is a more effective antiemetic regimen than ondansetron (OND) alone in children receiving chemotherapy. PATIENTS AND METHODS: Children who had solid tumors and who were receiving highly emetogenic chemotherapy, including cisplatin, carboplatin, cyclophosphamide, and ifosfamide, were randomized (1:1) in a double-blind fashion to receive either OND 0.15 mg/kg intravenously (i.v.) 30 min before and 4 and 8 h after chemotherapy and placebo, or OND + DEX (same OND doses plus DEX 8 mg/m2 i.v. 30 min before chemotherapy, followed by 16 mg/m2 in divided doses) as antiemetics. The patients were crossed over to the other antiemetic regimen when receiving an identical course of chemotherapy. Patients were monitored for emetic episodes, nausea, appetite, sense of well-being, and antiemetic adverse events. RESULTS: A total of 33 patients were evaluated. Sixty-one percent of the patients receiving OND + DEX regimens had a complete response for emetic episodes as compared with 23% with OND alone. Combined complete and major responses (two or less emetic episodes) were 86% for OND + DEX and 67% for OND. Failure for emetic episodes (more than five vomitings/day) were seen only in 7-10% of the total population. Minimal or no nausea was experienced in 74% of OND + DEX courses and in 52% of the OND courses. Appetite was better in OND + DEX courses (p = 0.006). Both antiemetic arms had similar adverse events. Mild to moderate sedation occurred in about half of the courses, followed by restlessness (29%), headache (17%), diarrhea (17%), and hiccups (2%). CONCLUSION: The combination of ondansetron and dexamethasone is superior to ondansetron alone to control emetic episodes in children receiving highly emetogenic chemotherapy (p = 0.04).

Adolescent↗

PubMed1994

Possible role of apamin-sensitive K+ channels in myotonic dystrophy.

Myotonic muscular dystrophy is a genetic disease characterized mainly by muscle atrophy and myotonia, a repetitive electrical activity of muscle. In the present study, the possible role of apamin-sensitive K+ channels in the genesis of myotonia was investigated. Apamin is a peptide from bee venom that specifically blocks small conductance Ca(2+)-activated K+ channels. The injection of a small amount of apamin (20-30 microliters, 10 mumol/L) into the thenar muscle of myotonic dystrophy patients decreased the basal electrical activity during the electromyogram in the 6 patients studied. Myotonic discharges after muscle percussion were more difficult to trigger and of smaller intensity and duration. In 2 controls and in 2 patients with generalized myotonia, as well as in 1 patient with myotonia congenita (where the defect is in chloride channels), apamin had no effect. These results suggest that apamin-sensitive K+ channels participate in the mechanism that generates myotonia in myotonic dystrophy.

Adult↗