Combination therapy of antibiotics and nanoparticles can be used against multi drug resistant microorganisms. Nanoparticles (NPs) have been reported to show antimicrobial activity. The antimicrobial activities of doped ZnO nanoparticles (ZnO NPs) were studied against fungi, gram-positive and gram-negative bacteria using the standard microdilution method. The interaction between the nanoparticle and the antibiotic was estimated by calculating the fractional inhibitory concentration (FIC index) of the combination through checkerboard assay. Experimental results demonstrated that 10% doped zinc oxide nanoparticles (ZnO NPs) exhibited the maximum antimicrobial effect in contrast with that of the 1% loading and pure ZnO nanoparticles. The enhancement in antimicrobial effect was seen when combined with antibiotic. Synergistic and additive effects were found. No antagonistic effect was found. More synergistic effect was observed when combined with ciprofloxacin than ampicillin. Fungus showed only additive effect. The results are quite in terms with MIC clearly depicting that high doping agent is most suitable for combined therapy. 100% synergistic interaction was observed in higher doping with both ciprofloxacin and ampicillin. This study provides a preliminary report of the synergistic activity of nanoparticles with antibiotics against different pathogenic strains. This provides groundwork for further studies on the combination therapy of nanoparticles with antibiotics.
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Antifungal susceptibility testing was performed on 197 yeast isolates from the BCCM/IHEM biomedical fungi and yeasts collection (Belgian Co-ordinated Collections of Micro-organisms / IPH-Mycology) to study the in vitro activity of voriconazole against fluconazole, itraconazole and amphotericin B. MICs of the four antifungal agents were determined by an adapted NCCLS M27-A microdilution reference method. MIC readings were visually and spectrophotometrically determined. Optical density data were used for calculation of the MIC endpoints. For amphotericin B, the MIC endpoint was defined as the minimal antifungal concentration that exerts 90% inhibition, compared to the control growth. The azoles endpoints were determined at 50% inhibition of growth. The MIC distribution of voriconazole susceptibilities showed that 193 isolates had a MIC < or = 2 microg/ml and 185 a MIC < or = 1 microg/ml. Cross-tabulation of voriconazole, fluconazole, and itraconazole MICs indicated that voriconazole MICs raised with fluconazole and itraconazole MICs. The in vitro data obtained in this study suggest that voriconazole may also be effective treating yeast infection in patients infected with fluconazole or itraconazole resistant isolates.
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Species distribution and antifungal susceptibility of Candida isolates at one institution were evaluated. Detection rates of fungi were examined for 5 years between 2007 and 2011. Sensitivities of fungi to amphotericin B, flucytosine, fluconazole, micafungin, itraconazole, and voriconazole were evaluated in blood culture-positive patients. A total of 3,832 fungal isolates were detected, including Candida albicans 66.5%, Candida glabrata 20.3%, Candida parapsilosis 6.2%, Candida tropicalis 5.5%, and others 1.5%. Candidemia was diagnosed in 131 patients, and C. albicans, C. parapsilosis, C. glabrata, C. tropicalis, and others were present in 42.0%, 27.5%, 16.0%, 8.4%, and 6.1% of these patients, respectively. Voriconazole had the lowest MIC90s against C. albicans and C. parapsilosis (0.015 and 0.25). Micafungin had a low MIC90 against C. glabrata and C. tropicalis. C. albicans was the most common fungus in patients with candidemia. Voriconazole and micafungin were effective against C. albicans. Amphotericin B was effective for C. parapsilosis, and micafungin showed good efficacy against C. glabrata and C. tropicalis.
Cryptococcus neoformans is a ubiquitous encapsulated yeast that causes significant infections which range from asymptomatic pulmonary colonization to the life threatening meningoencephalitis, especially in immunocompromised individuals. Cryptococcal meningitis is one of the AIDS-defining illnesses. Recent data have indicated that, the incidence of the cryptococcal infection is high in developing countries like India. We conducted this study to find out the incidence of cryptococcosis in this area.
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Candida biofilm formation is common during infection and environmental growth. We tested the impacts of three biocides (ethanol [EtOH], H(2)O(2), and sodium dodecyl sulfate) on Candida albicans, C. parapsilosis, and C. glabrata biofilms. Higher concentrations of the biocides were required for efficacy against biofilms than for efficacy against planktonic controls. A combination study with two biocides (EtOH and H(2)O(2)) and fluconazole demonstrated that the combination had enhanced efficacy.
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Trichosporon yeasts constitute emerging pathogens, implicated in organ-specific and systemic infections. In this first, comprehensive study of Trichosporon clinical isolates in Greece, 42 isolates were identified by sequencing the hypervariable D1/D2 domain of the Large Subunit (LSU) rDNA gene, while Trichosporon asahii were genotyped by sequencing the Intergenic Spacer 1 region, and antifungal susceptibilities were determined by the EDef 7.2 (EUCAST) method, in parallel with the CLSI standard. Trichosporon asahii was the primary species (37 isolates) followed by Trichosporon coremiiforme, Trichosporon dermatis, Trichosporon loubieri and Trichosporon mycotoxinivorans. One strain remained unidentified. Seven T. asahii genotypes were recorded. The major genotypes were: genotypes 4 (29%) and 3 (26%) followed by 1, 5 and 7 (9.5% each). Two novel genotypes were identified designated as 10 and 11. EUCAST MIC ≥2 mg/L was recorded in 58% of the isolates (amphotericin B), 41% (itraconazole), 41% (posaconazole) and 38% (voriconazole). Fluconazole MICs of ≥32 mg/L were recorded in 23.8% of the isolates. Analysis of variance performed on absolute values showed that the amphotericin B, itraconazole, posaconazole and voriconazole MICs of T. asahii were equivalent. Typically higher MIC values were displayed by fluconazole. Antifungal susceptibilities of the seven different genotypes were homogeneous. Agreements between EUCAST and CLSI ranged from 88.1 to 97.62%. Overall, the high MICs recorded among the Trichosporon isolates for all tested drugs justify routine susceptibility testing of clinical isolates.
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The clinical manifestations of histoplasmosis included fever, productive cough, chest pain, and abdominal pain, accompanied occasionally by neurological symptoms, lymph node enlargement or surface mass. Seven out of the 14 o patients had underlying immunosuppressive conditions, 9 had chest imaging changes, and 2 showed reduced white blood cells, red blood cells and platelets. The cases were initially diagnosis as tuberculosis, malignant tumor, or malignant lymphoma before the definite diagnosis was established pathologically. Ten patients received treatments with itraconazole, amphotericin B, fluconazole or voriconazole, and 9 of them responded favorably to the treatments.
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CYTOMEGALOVIRUS INFECTION: Primary prophylaxis is logical in high-risk subjects (CD4 < 50) who are asymptomatic; per os ganciclovir is under evaluation. Systematic secondary prophylaxis relies on intravenous ganciclovir or foscarnet. HERPES SIMPLEX AND ZOSTER: Secondary prevention with acyclovir is not recommended due to the risk of resistance. Herpes zoster retinitis is an absolute indication for continuous oral treatment with acyclovir. CRYPTOCOCCUS: Oral fluconazol is required for secondary prophylaxis. CANDIDIASIS: Due to the risk of resistant strains, neither primary nor secondary prophylasis is recommended. HISTIOPLASMOSIS: Risk of recurrence justifies secondary prophylaxis with itraconazole or fluconazole. COCCIDIODOMYCOSIS: Secondary prophylaxis with amphotericin B or fluconazole is mandatory. ASPERGILLOSIS: Itraconazole is the best agent when prophylaxis is needed.
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Saccharomyces cerevisiae pell and crd1 mutants deficient in the biosynthesis of mitochondrial phosphatidylglycerol (PG) and cardiolipin (CL) as well as Kluyveromyces lactis mutants impaired in the respiratory chain function (RCF) containing dysfunctional mitochondria show altered sensitivity to metabolic inhibitors. The S. cerevisiae pell mutant displayed increased sensitivity to cycloheximide, chloramphenicol, oligomycin and the cell-wall perturbing agents caffeine, caspofungin and hygromycin. On the other hand, the pel1 mutant was less sensitive to fluconazole, similarly as the K. lactis mutants impaired in the function of mitochondrial cytochromes. Mitochondrial dysfunction resulting either from the absence of PG and CL or impairment of the RCF presumably renders the cells more resistant to fluconazole. The increased tolerance of K. lactis respiratory chain mutants to amphotericin B, caffeine and hygromycin is probably related to a modification of the cell wall.
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The mating type locus (MTL) of Candida albicans contains the mating type genes and has, therefore, been assumed to play an exclusive role in the mating process. In mating-incompetent a/α cells, two of the mating type genes, MTLa1 and MTLα2, encode components of the a1-α2 corepressor that suppresses mating and switching. But the MTL locus of C. albicans also contains three apparently unrelated "nonsex" genes (NSGs), PIK, PAP and OBP, the first two essential for growth. Since it had been previously demonstrated that deleting either the a/α copy of the entire MTL locus, or either MTLa1 or MTLα2, affected virulence, we hypothesized that the NSGs in the MTL locus may also play a role in pathogenesis. Here by mutational analysis, it is demonstrated that both the mating type and nonsex genes in the MTL locus play roles in a/α biofilm formation, and that OBP is essential for impermeability and fluconazole resistance.
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There seems to be no survival benefit of antifungal agents given prophylactically or empirically to patients with cancer complicated by neutropenia. These agents should be restricted to patients with proved infection and those in randomised trials. A large, definitive placebo controlled trial of amphotericin B is needed.
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The four patients were hospitalized in the adult surgical intensive care unit. A total of 8 isolates (two per patient) from blood and catheter tip were analyzed.
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The azole antifungal fluconazole possesses only fungistatic activity in Candida albicans and, therefore, this human pathogen is tolerant to this agent. However, tolerance to fluconazole can be inhibited when C. albicans is exposed to fluconazole combined with the immunosuppressive drug cyclosporin A, which is known to inhibit calcineurin activity in yeast. A mutant lacking both alleles of a gene encoding the calcineurin A subunit (CNA) lost viability in the presence of fluconazole, thus making calcineurin essential for fluconazole tolerance. Consistent with this observation, tolerance to fluconazole was modulated by calcium ions or by the expression of a calcineurin A derivative autoactivated by the removal of its C-terminal inhibitory domain. Interestingly, CNA was also essential for tolerance to other antifungal agents (voriconazole, itraconazole, terbinafine, amorolfine) and to several other metabolic inhibitors (caffeine, brefeldin A, mycophenolic acid, fluphenazine) or cell wall-perturbing agents (SDS, calcofluor white, Congo red), thus indicating that the calcineurin pathway plays an important role in the survival of C. albicans in the presence of external growth inhibitors. Several genes, including PMC1, a vacuolar calcium P-type ATPase, were regulated in a calcineurin- and fluconazole-dependent manner. However, PMC1 did not play a direct role in the survival of C. albicans when exposed to fluconazole. In addition to these different properties, calcineurin was found to affect colony morphology in several media known to modulate the C. albicans dimorphic switch. In particular, calcineurin was found to be essential for C. albicans viability in serum-containing media. Finally, calcineurin was found to be necessary for the virulence of C. albicans in a mice model of infection, thus making calcineurin an important element for adequate adaptation to the conditions of the host environment.
The interaction of Cryptococcus neoformans with phagocytic cells of the innate immune system is a key step in disseminated disease leading to meningoencephalitis in immunocompromised individuals. Transcriptional profiling of cryptococcal cells harvested from cell culture medium or from macrophages found differential expression of metabolic and other functions during fungal adaptation to the intracellular environment. We focused on the ACL1 gene for ATP-citrate lyase, which converts citrate to acetyl-CoA, because this gene showed elevated transcript levels in macrophages and because of the importance of acetyl-CoA as a central metabolite. Mutants lacking ACL1 showed delayed growth on medium containing glucose, reduced cellular levels of acetyl-CoA, defective production of virulence factors, increased susceptibility to the antifungal drug fluconazole and decreased survival within macrophages. Importantly, acl1 mutants were unable to cause disease in a murine inhalation model, a phenotype that was more extreme than other mutants with defects in acetyl-CoA production (e.g. an acetyl-CoA synthetase mutant). Loss of virulence is likely due to perturbation of critical physiological interconnections between virulence factor expression and metabolism in C. neoformans. Phylogenetic analysis and structural modelling of cryptococcal Acl1 identified three indels unique to fungal protein sequences; these differences may provide opportunities for the development of pathogen-specific inhibitors.
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Candidaemia and other forms of invasive candidiasis (C/IC) are serious and costly events for hospitalized patients, particularly those in the ICU. Both fluconazole and the echinocandins are recommended as first-line therapy for C/IC. Resource use and cost considerations are important in selecting appropriate treatment but little information is available on the economic implications of using echinocandins in this setting.
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Evaluate the in vitro activity of anidulafungin in comparison to amphotericin B and fluconazole against different isolates of C. guilliermondii, and their efficacy in an immunosuppressed murine model of disseminated infection.
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Candidaemia is one of the leading causes of nosocomial bloodstream infections. There is a rise in the incidence of non-albicans candidaemia and emergence of anti-fungal resistance. We performed a retrospective laboratory-based study over a period of 2 years (January 2009 to December 2010) at our quaternary care multi super-specialty hospital in Southern India. There had been 68 Candida isolates detected from the bloodstream of 55 patients during the study period. Overall, 74% of cases were due to non-albicans Candida. C. tropicalis was most commonly isolated (39.7%), followed by C. albicans (26.4%). All Candida isolates remain susceptible to voriconazole, whereas highest degree of resistance was observed for fluconazole.
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The objective of the present investigation was to develop and evaluate microemulsion based gel for the vaginal delivery of fluconazole (FLZ). The solubility of FLZ in oils and surfactants was evaluated to identify components of the microemulsion. The ternary diagram was plotted to identify the area of microemulsion existence. Various gelling agents were evaluated for their potential to gel the FLZ microemulsion without affecting its structure. The bioadhesive potential and anti-fungal activity of the FLZ microemulsion based gel (FLZ-MBG) was determined in comparison to the marketed clotrimazole gel (Candid V gel) by in vitro methods. The vaginal irritation potential of the FLZ-MBG was evaluated in rabbits. The clinical efficacy of the FLZ-MBG and Candid V gel was evaluated in females suffering from vaginal candidiasis. The FLZ microemulsion exhibited globule size of 24 nm and polydispersity index of 0.98. Carbopol ETD 2020 could successfully gel the FLZ microemulsion without disturbing the structure. The FLZ-MBG showed significantly higher (P<0.05) in vitro bioadhesion and anti-fungal activity as compared to that of Candid V gel. The FLZ-MBG did not show any signs of vaginal irritation in the rabbits. The small-scale clinical studies indicated that the FLZ-MBG shows faster onset of action than Candid V gel although no difference was observed in the clinical efficacy.
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Distinguishing life-threatening toxoplasmic encephalitis (TE) from brain lymphoma in patients with acquired immunodeficiency syndrome (AIDS) may be difficult. Empiric anti-toxoplasmosis treatment is often initiated because of the reluctance in performing brain biopsies, which may delay the diagnosis and treatment of brain lymphoma in Japan. In this study, we retrospectively examined the clinical characteristics of 13 AIDS patients with TE in Japan, including magnetic resonance imaging and thallium 201 (201TI) single photon emission computed tomography (SPECT) findings, cerebral spinal fluid analysis, serology, and polymerase chain reaction (PCR) results. All patients improved on anti-toxoplasmosis treatment. Of the 11 patients who underwent serological testing, 6 (55%) had a positive serological result. Of the 7 patients who underwent PCR testing, 3 (42.9%) had a positive PCR result. Nine of 11 patients with TE (81.8%) had multiple lesions. Analysis of the sites of TE lesions did not reveal a difference in site predilection between TE and brain lymphoma. Uptake was negative in all 9 patients who underwent 201Tl SPECT. The study findings suggest that toxoplasma serostatus and PCR may be used to discriminate TE from brain lymphoma. No focal accumulation of 201TI is strongly suggestive of TE in patients with AIDS in Japan.
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Out of 319 episodes, 45 (14.1%) were breakthrough candidemia. Breakthrough candidemia occurred in patients with more acutely ill conditions, and the majority was caused by non-albicans Candida species (73.3%; 33 episodes). A total of 79.1% of breakthrough candidemia were caused by antifungal-susceptible Candida isolates and emergence of resistance was the mechanism in five cases of patients receiving fluconazole. Episodes of breakthrough candidemia had significantly higher illness severity and higher rates of fungemia-attributable mortality.
Fungal infection was detected in 36% of our patients. The independent risk factors associated with it were hypotension at hospitalization and prolonged antibiotic therapy. Antifungal therapy improved their chances of survival.
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A study of candidaemia in neonatal intensive care unit (NICU) over a 12-year period (1995-2006) taking into consideration demographic variables, risk factors, aetiological Candida species and therapeutic outcomes is presented. The yeast isolates were identified by VITEK2 yeast identification system and antifungal susceptibility was determined by E-test. Of 4815 neonates admitted in NICU, 182 cases of candidaemia were detected with an overall prevalence of 4.0% and crude mortality of 27.7%. The annual rate of candidaemia per 1000 admissions was the highest in 1997 (84 cases) and the lowest in 2004 (10 cases). Of the 112 assessable candidaemia cases, 78 (70%) occurred in very low birth weight neonates (< or =1500 g), 65 (58%) were born with gestational age of < or =30 weeks. The main identifiable risk factors were use of > or =2 antibiotics (87%), total parenteral nutrition for >5 days (82%), placement of central venous catheter (78%) and prior colonisation with Candida species (54%). Candida albicans and non-albicans Candida species accounted for 43% and 57% of candidaemia cases, respectively, and C. parapsilosis emerged as a predominant species. No fluconazole resistance was observed in C. albicans and C. parapsilosis isolates. This is the first comprehensive study on the epidemiology of neonatal candidiasis in Kuwait.
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A microdilution system was established for testing the susceptibility of Candida species to fluconazole and itraconazole. The assay used a sodium phosphate-buffered (0.1 mol l-1) Casitone/glucose medium (pH 7.2) supplemented with potassium, iron, magnesium, trace elements and vitamins. Tests were read photometrically after 24 h, and the minimum inhibitory concentration (MIC) was defined as the IC90. In nearly all strains sharp end points were observed. The MICs against Candida species without any known pre-exposure to azoles were found to range from 0.2 to 1.56 micrograms ml-1 for fluconazole and from 2.3 to 12 ng ml-1 for intraconazole. For strains from candidosis patients refractory to treatment with fluconazole the MICs of fluconazole ranged from 6.25 to 100 micrograms ml-1, while those for itraconazole varied between 12 and > 450 ng ml-1. The strains did not respond congruent to both azoles. A similar disparity of the MICs was observed with Candida tropicalis and Candida krusei. The unusually low MICs of itraconazole were attained because (1) the dilution series was prepared from a preformed (concentrated) dilution series in 75% dimethylsulphoxide and not directly by serial dilution in the broth and (2) the incubation was performed in microtitre plates made of quartz glass rather than in the generally used polystyrene microtitre plates.
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A promising approach for the eradication of biofilm formed by the yeast Candida albicans seems to be photodynamic inactivation (PDI). This work presents a use of methylene blue (MB, 1 mM) irradiated with a red laser (output power 190 mW/cm(2), wavelength 660 nm) for the eradication of a biofilm formed by the fluconazole-resistant (FLC-resistant) strain C. albicans CY 1123 compared to the standard strain C. albicans SC5314. The periods of irradiation corresponded to the fluence of 15, 23 and 57 J/cm(2) Effectiveness of PDI was evident with following percentage of survived biofilm cells: 24.57, 23.46, and 22.29% for SC5314 and 40.28, 17.91, and 5.89% for CY 1123, respectively, compared to the samples without irradiation. Light and confocal laser scanning microscopy confirmed the effectiveness of PDI. However, the morphological form of C. albicans seems to play an important role as well, since prolonged duration of irradiation did not increase efficiency of PDI on C. albicans SC5314. An experiment with the yeast-to-hyphae transition revealed that the FLC-resistant strain expressed a markedly reduced capacity to form hyphae compared to SC5314. We summarized that PDI was effective on biofilm formed by the FLC-resistant strain, but resistance most likely did not play significant role in PDI. Additionally, we observed differences in susceptibility to PDI between biofilms composed of the mycelia and only of the yeasts, and finally, the employment of a laser in PDI enabled a decreasing period of irradiation while maintaining the high effectiveness of PDI.
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The broth macrodilution method (BMM) for antifungal susceptibility testing, approved by the National Committee for Clinical Laboratory Standards (NCCLS), was found to have deficiencies in testing of the antifungal activity of a new type of antifungal agent, a nonantibacterial chemically modified tetracycline (CMT-3). The high content of phosphate in the medium was found to greatly increase the MICs of CMT-3. To avoid the interference of phosphate in the test, a new method using potato dextrose agar (PDA) as a culture medium was developed. Eight strains of fungi, including five American Type Culture Collection strains and three clinical isolates, were used to determine the MICs of amphotericin B and itraconazole with both the BMM and the PDA methods. The MICs of the two antifungal agents determined with the PDA method showed 99% agreement with those determined with the BMM method within 1 log(2) dilution. Similarly, the overall reproducibility of the MICs with the PDA method was above 97%. Three other antifungal agents, fluconazole, ketoconazole, and CMT-3, were also tested in parallel against yeasts and molds with both the BMM and the PDA methods. The MICs of fluconazole and ketoconazole determined with the PDA method showed 100% agreement within 1 log(2) dilution of those obtained with the BMM method. However, the MICs of CMT-3 determined with the BMM method were as high as 128 times those determined with the PDA method. The effect of phosphate on the antifungal activity of CMT-3 was evaluated by adding Na2HPO4 to PDA in the new method. It was found that the MIC of CMT-3 against a Penicillium sp. increased from 0.5 microg/ml (control) to 2.0 microg/ml when the added phosphate was used at a concentration of 0.8 mg/ml, indicating a strong interference of Na2HPO4 with the antifungal activity of CMT-3. Except for fluconazole, all the other antifungal agents demonstrated clear end points among the yeasts and molds tested. Nevertheless, with its high reproducibility, good agreement with NCCLS proposed MIC ranges, and lack of interference of phosphate, the PDA method shows promise as a useful assay for antifungal susceptibility testing and screening for new antifungal agents, especially for drugs that may be affected by high (supraphysiologic) phosphate concentrations.
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Infectious esophagitis can have significant implications in an impaired host. Described most commonly in immunocompromised patients, infectious esophagitis can also occasionally be discovered in immunocompetent individuals in several unique clinical settings. Evaluation of the typical presenting complaints, such as dysphagia or odynophagia, are especially important in immunocompetent patients, and therapy should be directed at the appropriate predisposing condition and resultant infectious agent. In immunocompromised patients, however, clinical experience supports the use of empiric therapy in patients without concomitant systemic complaints. Especially in AIDS patients or those with lymphoma or leukemia, the initial approach to infectious esophagitis complaints (ie, dysphagia or odynophagia) is to begin an empiric trial of oral systemic fluconazole for presumed candidal esophagitis. If the individual remains symptomatic after 3 to 7 days or has any associated systemic complaints or concerning clinical findings (eg, hematemesis), then upper endoscopy with biopsies is indicated. If an etiologic agent other than Candida is defined by histologic, immunohistochemical, or culture methods, then appropriate therapy can be initiated. There are many important and pathologic agents implicated in infectious esophagitis. Thus, directed therapy needs to be administered appropriately and in a timely fashion to avoid poor short-term problems or long-term sequelae.
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Of the 769 patients, pulmonary TB only was diagnosed in 461 (60%), both pulmonary and extrapulmonary TB in 78 (10%), extrapulmonary TB at one site in 223 (29%), and extrapulmonary TB at more than one site in seven (1%) patients. Death during TB treatment occurred in 59 of 308 patients (19%) with any extrapulmonary involvement. In a proportional hazards model, patients with extrapulmonary TB had an increased risk of death if they had meningitis, and a CD4+ T-lymphocyte count <200 cells/microl. Patients who received co-trimoxazole, fluconazole, and antiretroviral therapy during TB treatment had a lower risk of death.
Retrospective, randomized, controlled clinical studies were reviewed.