Biomedical subjects

Brian Wong

Publications and source records attributed to Brian Wong.

14 recordsLinked to original sources

PubMed2006

Measurement of serum D-arabinitol/creatinine ratios for initial diagnosis and for predicting outcome in an unselected, population-based sample of patients with Candida fungemia.

D-Arabinitol (DA) is a useful diagnostic marker for candidiasis in patients with neutropenia and other high-risk groups, but its use in unselected patients with a broad range of underlying diseases and conditions has not been studied. We used an automated enzymatic fluorometric assay to measure serum DA/creatinine ratios (DA/cr's) in 30 healthy adults, 100 hospitalized controls without Candida fungemia, and 83 patients from a study of all Candida fungemias in Connecticut between October 1998 and September 1999. Sixty-three of 83 (76%) fungemic patients and 11 of 100 (11%) nonfungemic controls had serum DA/cr's >or=3.9 microM/mg/dl (mean + 3 standard deviations for 30 healthy adults). High serum DA/cr's were less frequent in patients with cancer or fungemia caused by the DA nonproducer Candida glabrata than in patients with cancer or fungemia caused by a DA producer, C. albicans, C. tropicalis, or C. parapsilosis. The serum DA/cr was first >or=3.9 microM/mg/dl before, on the same day as, or after the first positive blood culture was drawn for 30 (36%), 22 (27%), and 11 (13%) fungemia patients, respectively. Mortality did not differ significantly among the patients with high or normal initial or peak serum DA/cr's, but mortality was higher if any serum DA/cr value was >or=3.9 microM/mg/dl 3 or more days after the onset of fungemia (18/27 versus 4/24 patients, respectively; P < 0.001). We conclude that serum DA/cr's are useful both for the initial diagnosis of Candida fungemia and for prognostic purposes for unselected patients with a broad range of underlying diseases and conditions.

Adolescent↗

PubMed2005

Shape retention in porcine and rabbit nasal septal cartilage using saline bath immersion and Nd:YAG laser irradiation.

BACKGROUND AND OBJECTIVES: The process of altering the shape of cartilage using heat has been referred to as thermoforming, and presents certain clinical benefits in reconstructive surgical procedures within the head and neck. Thermoforming allows cartilage in the upper airway and face to be reshaped without the use of classic surgical maneuvers such as carving, morselizing, or suturing. The goal of this study was to determine the dependence of cartilage shape change on both temperature and laser dosimetry using two thermoforming methods: saline bath immersion and laser irradiation. STUDY DESIGN/MATERIALS AND METHODS: Ex-vivo rabbit and porcine nasal septal cartilages were mechanically deformed and reshaped using the two thermoforming methods. With saline bath immersion using rabbit cartilage, each specimen was deformed by securing it to a small copper tube (outer diameter 8 mm) using dental bands. For porcine cartilage immersed in a saline bath, each sample was mechanically deformed between two pieces of wire mesh attached to a semicircular acrylic block. With both porcine and rabbit cartilage, the specimen and apparatus were then immersed in a hot saline bath for time intervals varying from 20 and 320 seconds and at constant temperatures between 62 and 74 degrees C. In laser reshaping, the cartilage specimens were mechanically deformed on a jig and consecutively irradiated with an Nd:YAG laser (lambda = 1.32 microm) in several spots for 6-16 seconds and irradiances of 10.2-40.7 W/cm2 per spot. After either saline bath heating or irradiation, cartilage specimens were immersed in room temperature saline for 15 minutes, then upon removal from the jig the length between the ends of each specimen was measured in order to calculate the resulting bend angle. RESULTS: The transition zone for cartilage reshaping was defined as where a significant increase in bend angle was observed between consecutive times of immersion/irradiation at the same temperature/irradiance. For the saline bath experiments, the transition zone was observed between 59-68 degrees C and 62-68 degrees C for porcine and rabbit cartilage, respectively. Similar transition zones occurred with laser irradiation below irradiances of 20.4 W/cm2 for both porcine and rabbit cartilage. In addition, the dosimetry pairs in the transition zones produce peak temperatures below the thresholds determined from the saline bath immersion studies. CONCLUSIONS: The critical transition temperature region was determined by the sharp increase in bend angle at consecutive times of immersion at the same temperature. This range was determined to be 59-68 degrees C and 62-68 degrees C for porcine and rabbit cartilage, respectively. Similar transition zones for dosimetry occurred below 20.4 W/cm2 during cartilage irradiation in both species.

Animals↗

PubMed2005

Intracellular trafficking of fluorescently tagged proteins associated with pathogenesis in Candida albicans.

Proteins that enter the secretory pathway play important roles in virulence and pathogenesis in Candida albicans, but our understanding of the trafficking of these proteins is in its early stages. In Saccharomyces cerevisiae, dominant negative alleles of YPT1 and SEC4 interrupt secretory traffic at pre- and post-Golgi steps, respectively. We therefore used a dominant negative genetic approach to examine the intracellular trafficking of several proteins associated with virulence or azole resistance. When the dominant negative ypt1(N121I) allele of C. albicans was overexpressed, yellow-fluorescent protein (YFP) tagged forms of two plasma membrane transporters (Cdrlp and Ftrlp) and the vacuolar membrane ABC transporter Mltlp accumulated in intracellular structures that appeared related to the ER, but localization of Cdc10p and Int1p was unaffected. When the dominant negative sec4(S28N) allele of C. albicans was overexpressed, Cdrlp and Ftrlp accumulated intracellularly, and localization of Mltlp, Cdc10p and Int1p was unaffected. These results imply that (i) Cdrlp and Ftrlp are transported to the plasma membrane by the general secretory pathway, (ii) Mlt1p enters the secretory pathway but is diverted to the vacuole at an early post-Golgi step, and (iii) like Cdc10p, Int1p does not enter the general secretory pathway.

ATP-Binding Cassette Transporters↗

PubMed2005

Comparison of flexible ureteroscopes: deflection, irrigant flow and optical characteristics.

PURPOSE: We measured and compared the deflection, irrigation flow rates, distortion, resolution and light transmission of new generation flexible ureteroscopes. MATERIALS AND METHODS: Multiple characteristics of 5 flexible ureteroscopes (ACMI DUR-8 Elite, Olympus URF-P3, Storz 11278AU1 [Flex-X], Wolf 7330.072 and Wolf 7325.172) commonly available in the market were measured and compared. Measured data included active deflection, irrigation flow rates and optical characteristics. Each ureteroscope was evaluated with an empty working channel and with various accessories. Optical characteristics, specifically resolution and distortion, were measured using test targets (Edmund Optics, Barrington, New Jersey). Light transmission was also measured from the ureteroscope tip at 50% and 100% intensity. All 5 flexible ureteroscopes were tested in a laboratory setting using a Storz OR 1 system to capture the images. RESULTS: For all 5 ureteroscopes the angle of deflection was most impaired by a 365 microm laser fiber probe and least impaired by a 2.2Fr nitinol basket. Among all 5 ureteroscopes irrigation flow rate was most impaired with a 3.0Fr basket and least impaired with 200 microm laser fiber. The Wolf 7325.172 had the highest observed resolution of 25.39 lines per mm and the Wolf 7330.072 had the lowest distortion at 11.9%. The Karl Storz Flex-X and the ACMI DUR-8 Elite had the highest light output at 374 and 364 mV, respectively. CONCLUSIONS: The various flexible ureteroscopes differ with regard to flow rates as well as degree of deflection with either an empty or an occupied working channel. The Wolf flexible ureteroscope with a slightly larger working channel and a fused quartz bundle provided for superior flow and better optical performance. However, the greatest amount of tip deflection and highest light output were found in the ACMI and Karl Storz flexible ureteroscopes.

Artifacts↗

PubMed2005

The MAP kinase Hog1p differentially regulates stress-induced production and accumulation of glycerol and D-arabitol in Candida albicans.

Candida albicans produces and accumulates large amounts of the polyols d-arabitol and glycerol in culture, and/or in infected mammalian tissues. However, the effects of environmental stresses on production and accumulation of these polyols, and the means by which polyol production and accumulation are regulated have not been studied. C. albicans grown in glucose at 30 degrees C (i) produced maximal amounts of glycerol within 6 h, (ii) produced maximal amounts of d-arabitol and ribitol within 12 h, and (iii) released most of these polyols into the extracellular environment. C. albicans responded to osmotic and citric acid stress by producing and accumulating more glycerol, and to temperature and oxidative stresses by producing more d-arabitol. The increase in intracellular glycerol was proportional to extracellular osmolarity, suggesting that glycerol functions as an osmolyte. The MAP kinase Hog1p is required for wild-type glycerol production in several fungal species subjected to osmotic stress, but it is not known if Hog1p plays a role in regulating d-arabitol production. Therefore, two C. albicans hog1 null mutants were constructed and tested for the ability to produce glycerol and d-arabitol in response to environmental stresses. The ability to grow and produce glycerol when exposed to osmotic or citric acid stresses, and to produce d-arabitol when exposed to oxidative stress, was partially dependent on Hog1p, but the ability to produce d-arabitol when exposed to temperature stress was Hog1p independent. These results imply that multiple pathways regulate glycerol and d-arabitol synthesis in C. albicans.

Adaptation, Physiological↗

PubMed2003

An analysis of the Candida albicans genome database for soluble secreted proteins using computer-based prediction algorithms.

We sought to identify all genes in the Candida albicans genome database whose deduced proteins would likely be soluble secreted proteins (the secretome). While certain C. albicans secretory proteins have been studied in detail, more data on the entire secretome is needed. One approach to rapidly predict the functions of an entire proteome is to utilize genomic database information and prediction algorithms. Thus, we used a set of prediction algorithms to computationally define a potential C. albicans secretome. We first assembled a validation set of 47 C. albicans proteins that are known to be secreted and 47 that are known not to be secreted. The presence or absence of an N-terminal signal peptide was correctly predicted by SignalP version 2.0 in 47 of 47 known secreted proteins and in 47 of 47 known non-secreted proteins. When all 6165 C. albicans ORFs from CandidaDB were analysed with SignalP, 495 ORFs were predicted to encode proteins with N-terminal signal peptides. In the set of 495 deduced proteins with N-terminal signal peptides, 350 were predicted to have no transmembrane domains (or a single transmembrane domain at the extreme N-terminus) and 300 of these were predicted not to be GPI-anchored. TargetP was used to eliminate proteins with mitochondrial targeting signals, and the final computationally-predicted C. albicans secretome was estimated to consist of up to 283 ORFs. The C. albicans secretome database is available at http://info.med.yale.edu/intmed/infdis/candida/

Algorithms↗

PubMed2003

Does 24-hour ST-segment resolution postfibrinolysis add prognostic value to a Q wave? An ASSENT 2 electrocardiographic substudy.

BACKGROUND: Both ST resolution and Q-wave development postfibrinolysis provide important prognostic insights in patients with acute myocardial infarction (MI). However, the relative contributions of these 2 factors to risk assessment have not been examined prospectively. METHODS AND RESULTS: ST resolution and Q development were evaluated 24 to 36 hours (24-36 h) postfibrinolysis in ASSENT-2: 13,100 out of 16,949 patients who had both baseline and 24-36 h electrocardiograms free of confounders (left bundle branch block, ventricular rhythm, reinfarction before 24-36 h electrocardiograms) were included in this analysis. Q-wave MI evolved in 10,466 patients (79.9%) and 2634 patients (20.1%) had non-Q-wave MI at 24-36 h postfibrinolysis. Mortality rates at 1-year were 7.0% for patients with Q-wave MI and 5.8% for non-Q-wave MI patients, respectively (P =.046). Patients with Q-wave MI versus those without were less likely to have complete ST-segment resolution (49.1% vs 59.1%) and more likely to have partial (37.1% vs 27.8%) or no resolution (13.8% vs 13.1%) at 24 to 36 hours postfibrinolysis (P <.001). Mortality rates at 1 year for Q-wave MI with complete, partial, and no resolution were 5.2%, 8.1%, and 10.1%, respectively (P <.001), and for non-Q-wave MI with complete, partial, and no resolution were 4.5%, 7.6%, and 8.0% (P =.003). CONCLUSION: These results demonstrate the additional prognostic significance of ST-segment resolution to Q-wave development at 24 to 36 hours after fibrinolysis.

Aged↗

PubMed2003

Use of green fluorescent protein fusions to analyse the N- and C-terminal signal peptides of GPI-anchored cell wall proteins in Candida albicans.

Glycophosphatidylinositol (GPI)-anchored proteins account for 26-35% of the Candida albicans cell wall. To understand the signals that regulate these proteins' cell surface localization, green fluorescent protein (GFP) was fused to the N- and C-termini of the C. albicans cell wall proteins (CWPs) Hwp1p, Als3p and Rbt5p. C. albicans expressing all three fusion proteins were fluorescent at the cell surface. GFP was released from membrane fractions by PI-PLC and from cell walls by beta-glucanase, which implied that GFP was GPI-anchored to the plasma membrane and then covalently attached to cell wall glucans. Twenty and 25 amino acids, respectively, from the N- and C-termini of Hwp1p were sufficient to target GFP to the cell surface. C-terminal substitutions that are permitted by the omega rules (G613D, G613N, G613S, G613A, G615S) did not interfere with GFP localization, whereas some non-permitted substitutions (G613E, G613Q, G613R, G613T and G615Q) caused GFP to accumulate in intracellular ER-like structures and others (G615C, G613N/G615C and G613D/G615C) did not. These results imply that (i) GFP fusions can be used to analyse the N- and C-terminal signal peptides of GPI-anchored CWPs, (ii) the omega amino acid in Hwp1p is G613, and (iii) C can function at the omega+2 position in C. albicans GPI-anchored proteins.

Amino Acid Sequence↗

PubMed2002

Current status of nonculture methods for diagnosis of invasive fungal infections.

The incidence of invasive fungal infections has increased dramatically in recent decades, especially among immunocompromised patients. However, the diagnosis of these infections in a timely fashion is often very difficult. Conventional microbiologic and histopathologic approaches generally are neither sensitive nor specific, and they often do not detect invasive fungal infection until late in the course of disease. Since early diagnosis may guide appropriate treatment and prevent mortality, there has been considerable interest in developing nonculture approaches to diagnosing fungal infections. These approaches include detection of specific host immune responses to fungal antigens, detection of specific macromolecular antigens using immunologic reagents, amplification and detection of specific fungal nucleic acid sequences, and detection and quantitation of specific fungal metabolite products. This work reviews the current status and recent developments as well as problems in the design of nonculture diagnostic methods for invasive fungal infections.

Antibodies, Fungal↗

PubMed2001

Overexpression of a dominant-negative allele of YPT1 inhibits growth and aspartyl protease secretion in Candida albicans.

To investigate the pre-Golgi secretion pathway in the pathogenic yeast Candida albicans, we cloned the C. albicans homologue of the Saccharomyces cerevisiae protein secretion gene YPT1. The C. albicans YPT1 ORF contained a 624 bp intronless ORF encoding a deduced protein of 207 aa and 2.3 kDa. This deduced protein was 77% identical to S. cerevisiae Ypt1 protein (Ypt1p) and it contained GTP-binding domains that are conserved among all known ras-like GTPases. Multicopy plasmids containing C. albicans YPT1 complemented the temperature-sensitive S. cerevisiae ypt1 (A136D) mutation. One chromosomal YPT1 allele in C. albicans CAI4 was readily disrupted by homologous gene targeting, but attempts to disrupt the second allele yielded no viable null mutants. Since this suggested that C. albicans YPT1 may be essential, a mutant ypt1 allele was constructed encoding the amino acid substitution analogous to the N121I substitution in a known trans-dominant inhibitor of S. cerevisiae Ypt1p. Next, a GAL1-regulated plasmid was used to express the mutant ypt1(N121I) allele in C. albicans CAI4. Ten of 11 transformants tested grew normally in glucose and poorly in galactose, and plasmid curing restored growth to wild-type levels. When these transformants were incubated in galactose, secretion of aspartyl proteinase (Sap) was inhibited and membrane-bound secretory vesicles accumulated intracellularly. These results imply that C. albicans YPT1 is required for growth and protein secretion, and they confirm the feasibility of using inducible dominant-negative alleles to define the functions of essential genes in C. albicans.

Alleles↗

PubMed2000

Mannitol-1-phosphate dehydrogenase from Cryptococcus neoformans is a zinc-containing long-chain alcohol/polyol dehydrogenase.

Cryptococcus neoformans, the causative agent of cryptococcosis, produces large amounts of mannitol in culture and in infected mammalian hosts. Although there is considerable indirect evidence that mannitol synthesis may be required for wild-type stress tolerance and virulence in C. neoformans, this hypothesis has not been tested directly. It has been proposed that mannitol-1-phosphate dehydrogenase (MPD) is required for fungal mannitol synthesis, but no MPD-deficient fungal mutants or cDNAs or genes encoding fungal MPDs have been described. Therefore, C. neoformans was purified from a 148 kDa homotetramer of 36 kDa subunits that catalysed the reaction mannitol1-phosphate+NAD--><--fructose 6-phosphate+NADH. Partial peptide sequences were used to isolate the corresponding cDNA and gene, and the deduced MPD protein was found to be homologous to the zinc-containing long-chain alcohol/polyol dehydrogenases. Lysates of Saccharomyces cerevisiae transformed with the cDNA of interest (but not vector-transformed controls) contained MPD catalytic activity. Lastly, Northern analyses demonstrated MPD mRNA in glucose- and mannitol-grown C. neoformans cells. Thus, MPD has been purified and characterized from C. neoformans, and the corresponding cDNA and gene (MPD1) cloned and sequenced. Availability of C. neoformans MPD1 should permit direct testing of the hypotheses that (i) MPD is required for mannitol biosynthesis and (ii) the ability to synthesize mannitol is essential for wild-type stress tolerance and virulence.

Alcohol Oxidoreductases↗

PubMed1996

Stress tolerance and pathogenic potential of a mannitol mutant of Cryptococcus neoformans.

Cryptococcus neoformans produces large amounts of the acyclic hexitol mannitol in culture and infected animals, but the functional and pathogenic significance of mannitol production by this fungus is not known. We exposed C. neoformans H99 (Cn H99) to UV irradiation (1 x LD50) and screened survivors for mannitol production. A mutant, Cn MLP (Mannitol Low Producer), synthesized less mannitol from glucose (2.7 vs 8.2 nmol per 10(8) cells min-1 at 37 degrees C) and contained less intracellular mannitol (1 vs 11 mumol per 10(6) cells at 37 degrees C) than did Cn H99. Cn MLP and Cn H99 were similar with respect to carbon assimilation patterns, rates of glucose consumption, growth rates at 30 degrees C, urease and phenoloxidase activities, morphology, capsule formation, mating type, electrophoretic karyotype, rapid amplification of polymorphic DNA (RAPD) patterns and antifungal susceptibility. However, Cn MLP was more susceptible than was Cn H99 to growth inhibition and killing by heat and high NaCl concentrations. Also, the LD50 values in mice injected intravenously were 3.7 x 10(6) c.f.u. for Cn MLP compared to 6.9 x 10(2) c.f.u. for Cn H99. Moreover, 500 c.f.u. Cn H99 intravenously killed 12 of 12 mice by 60 d, whereas all mice given the same inoculum of Cn MLP survived. Classical genetic studies were undertaken to determine if these differences were due to a single mutation, but the basidiospores were nonviable. These results suggest that the abilities of C. neoformans to produce and accumulate mannitol may influence its tolerance to heat and osmotic stresses and its pathogenicity in mice.

Animals↗

PubMedDate not supplied

Cardiac plasmacytoma.

A 69-year-old white woman presented with a 3-month history of progressive dyspnea, orthopnea, fatigue and weakness. Clinical, diagnostic imaging and echocardiographic investigations suggested an occult primary cancer with metastasis to the heart. The patient's condition deteriorated gradually, and she died 2 months later. At autopsy, a malignant tumor encasing the heart and a 1-cm solitary tumor nodule in the lower lobe of the left lung were found. Histologic and electron microscopic studies revealed a plasmacytoma predominantly involving the epicardium and a small solitary plasmacytoma located in the left lung. The two tumors were further confirmed by immunohistochemical studies that showed monoclonal IgG expression and kappa light chain restriction.

Aged↗

PubMedDate not supplied

In vivo optical coherence tomography of the nasal mucosa.

BACKGROUND: Optical coherence tomography (OCT) is an emerging imaging modality that uses light to produce in vivo high-resolution cross-sectional images (7 microm) of tissues to depths of up to 3 mm. OCT is analogous to ultrasound, but relies on interferometry and low-coherence optical sources to produce images of tissue structure at the histological level. METHODS: In this study, OCT was used to image the mucosa overlying structures in the nasal cavity to obtain information regarding normative in vivo tissue microstructure. An OCT system using a Michaelson interferometer and a 1.3-microm broadband light source was incorporated into a fiber-optic imaging device that was inserted into the nasal cavity. Cross-sectional tomographic images of the anterior and posterior nasal septum, turbinates, and vestibule were acquired in 44 patients in either the office or the operating room during surgical endoscopy. RESULTS: OCT images of the nasal mucosa identified the distinct boundaries between the epithelium, lamina propria, and underlying bone/cartilaginous tissue. Within the lamina propria, features consistent with glands, ducts, and blood vessels were clearly identified. In patients who underwent decongestant therapy, before and after images showed distinct morphological changes in the mucosa. The thickness of the epithelium was tabulated, as well. CONCLUSION: This study establishes the potential of using OCT to produce high-resolution images of the nasal mucosa. As an in vivo tissue microstructural imaging modality, OCT may be valuable in studying the impact of allergic and infectious disease on the nasal mucosa and monitoring its response to pharmacologic therapy.

Adolescent↗