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AIM: The study aimed to evaluate the compressive strength and surface hardness of a type V dental stone after hypochlorite disinfection. MATERIALS AND METHODS: Two types of specimens were made according to the American Dental Association (ADA) specification no. 25 for each wet compressive strength, dry compressive strength, and surface hardness. The specimens were split into three groups with 30 samples each according to the type of disinfection. All specimens were immersed in their respective disinfecting solutions for 30 minutes at room temperature and after removal, they were left to dry for 24 hours at room temperature. Total five cycles of immersion and drying were followed. A compressive strength test was done using a universal testing machine. Wet compressive strength was tested one hour after the last cycle and dry compressive strength was tested 7 days after the last cycle. Surface hardness was measured after 48 hours using Vickers hardness test. The results were statistically analyzed. RESULTS: There was a statistical difference between the calcium hypochlorite and sodium hypochlorite groups for both dry and wet compressive strength. The mean wet compressive strength of calcium hypochlorite was higher when compared to the sodium hypochlorite group and it was statistically significant (p = 0.042). The results were similar and statistically significant (p = 0.003) for dry compressive strength. When the mean surface hardness of the sodium hypochlorite (As) group was compared to calcium hypochlorite the results were highly significant (p = 0.0001) with the mean surface hardness of the calcium hypochlorite group more than the sodium hypochlorite group. CONCLUSION: Calcium hypochlorite used as a disinfectant showed better compressive strength and surface hardness when compared to sodium hypochlorite as a disinfectant. CLINICAL SIGNIFICANCE: Dental casts poured in the contaminated impressions which might not be disinfected at all or properly. They also come in contact with the prosthesis that might be tried inside the patient's mouth and sent to a lab for corrections without disinfecting the cast causing cross-contamination between patients, dentists, and laboratory personnel. However, immersion disinfection with sodium or calcium hypochlorite might affect important properties of the cast. Any negative effect on the mechanical or physical properties of the cast will affect the final outcome of the prosthesis.
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Desinfectantes , Ácido Hipocloroso , Sulfato de Calcio , Fuerza Compresiva , Dureza , Humanos , Ensayo de Materiales , Modelos Dentales , Hipoclorito de Sodio/farmacología , Propiedades de SuperficieRESUMEN
INTRODUCTION: Human immunodeficiency virus (HIV) infection is a major worldwide health problem characterized by progressive immunosuppression. The morbidity of HIV patients is due to its association with opportunistic infections among which oral candidiasis is common. Regardless of HIV status, candidiasis can prevail when their immune system is depressed. Oral candidiasis can thus serve as a useful marker for both restoration of immune functions and HIV disease progression. Routine identification of Candida species is laborious and time-consuming. HICHROM agar stains different species into different colors facilitating rapid reliable identification of candida species as they differ in their virulence and sensitivity to antifungal drugs. MATERIALS AND METHODS: This cross-sectional study includes a total of 200 HIV seropositive patients from Anti-Retroviral Therapy Centre, West Godavari District. Unstimulated saliva samples were collected in a screw-capped universal container. Five microliters of each sample using a sterile inoculating loop is streaked on the chromogenic agar culture media. The colonies formed are counted using a magnifying glass and LAPIZ colony counter. Candidal colony-forming units per milliliter were analyzed, compared and correlated among different study groups. Different candida species were also identified in the study. RESULTS: Nonalbicans Candida was the most common species isolated in our study accounting for 53% and Candida albicans accounting for 47%. Considering initial and final CD4 counts, there is improvement in patients on retroviral therapy. CONCLUSION: Identification of the species is important for epidemiological reasons and for treatment purposes to ensure a better prognosis since some species present reduced susceptibility to azoles.
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T cells reacting to self-components can promote tissue damage when escaping tolerogenic control mechanisms which may result in autoimmune disease. The current treatments for these disorders are not antigen (Ag) specific and can compromise host immunity through chronic suppression. We have previously demonstrated that co-administration of encapsulated or free Ag with tolerogenic nanoparticles (tNPs) comprised of biodegradable polymers that encapsulate rapamycin are capable of inhibiting Ag-specific transgenic T cell proliferation and inducing Ag-specific regulatory T cells (Tregs). Here, we further show that tNPs can trigger the expansion of endogenous Tregs specific to a target Ag. The proportion of Ag-specific Treg to total Ag-specific T cells remains constant even after subsequent Ag challenge in combination with a potent TLR7/8 agonist or complete Freund's adjuvant. tNP-treated mice do not develop experimental autoimmune encephalomyelitis (EAE) after adoptive transfer of encephalitogenic T cells; furthermore, tNP treatment provided therapeutic protection in relapsing EAE that was transferred to naïve animals. These findings describe a potent therapy to expand Ag-specific Tregs in vivo and suppress T cell-mediated autoimmunity.
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Autoantígenos/inmunología , Encefalomielitis Autoinmune Experimental/inmunología , Tolerancia Inmunológica/inmunología , Inmunosupresores/administración & dosificación , Sirolimus/administración & dosificación , Linfocitos T Reguladores/inmunología , Animales , Femenino , Ratones , Nanopartículas , Vacunas Sintéticas/inmunologíaRESUMEN
INTRODUCTION: Oral cancer is a multistep process involving foul play of proto-oncogenes that induce cell proliferation, inactivation of tumor suppressor gene and cessation of programmed cell death. Among various proto-oncogenes, the nature and behavior of Bcl-2 and c-Myc in oral precancerous/cancerous lesions were obscured which require further assessment for better understanding of etiology, treatment and prognosis. AIM: The aim of the study is to assess the expression of Bcl-2 and c-Myc in oral epithelial dysplasia and oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: This retrospective study of 70 (oral dysplasia [30], OSCC [30] and normal gingiva [10]) is immunohistochemically assessed for Bcl-2 and c-Myc for distribution, intensity, percentage of positive cells, localization and immunoreactive scores using ImageJ software. RESULTS: Bcl-2 showed 60% and 37% positivity within dysplasia and OSCC, respectively (P = 0.03); c-Myc showed 87% and 90% positivity within dysplasia and OSCC, respectively. In OSCC, c-Myc showed moderate intensity (P = 0.04). Average percentage of positive cells expressing c-Myc and Bcl-2 increased proportionally within grades of dysplasia (P = 0.000 and P = 0.008, respectively), whereas in OSCC, only c-Myc showed significant expression (P = 0.021). Localization of c-Myc was seen in the nucleus among OSCC (P = 0.01). c-Myc and Bcl-2 showed moderate immunoreactivity in dysplasia (P = 0.005 and P = 0.013, respectively), whereas in OSCC, moderate immunoreactivity of c-Myc (P = 0.05) was observed. CONCLUSION: Variable expression of c-Myc and Bcl-2 reveals that these proteins act in synergism in early phases of carcinogenesis, whereas in later stages, due to the diminished activity of Bcl-2, c-Myc interacts incoordination with other oncogenes contributing to tumor progression.
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Mulberry (Morus species) leaf is the sole food for monophagous silkworms, Bombyx mori L. Abiotic stresses such as drought, salinity, and high temperature, significantly decrease mulberry productivity and post-harvest water loss from leaves influence silkworm growth and cocoon yield. Leaf surface properties regulate direct water loss through the cuticular layer. Leaf surface waxes, contribute for cuticular resistance and protect mesophyll cells from desiccation. In this study we attempted to overexpress AtSHN1, a transcription factor associated with epicuticular wax biosynthesis to increase leaf surface wax load in mulberry. Agrobacterium mediated in vitro transformation was carried out using hypocotyl and cotyledonary explants of Indian mulberry (cv. M5). Mulberry transgenic plants expressing AtSHN1 displayed dark green shiny appearance with increased leaf surface wax content. Scanning electron microscopy (SEM) and gas chromatograph-mass spectrometry (GC-MS) analysis showed change in pattern of surface wax deposition and significant change in wax composition in AtSHN1 overexpressors. Increased wax content altered leaf surface properties as there was significant difference in water droplet contact angle and diameter between transgenic and wild type plants. The transgenic plants showed significant improvement in leaf moisture retention capacity even 5 h after harvest and there was slow degradation of total buffer soluble protein in detached leaves compared to wild type. Silkworm bioassay did not indicate any undesirable effects on larval growth and cocoon yield. This study demonstrated that expression of AtSHN1, can increase surface wax load and reduce the post-harvest water loss in mulberry.
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The development of antidrug antibodies (ADAs) is a common cause for the failure of biotherapeutic treatments and adverse hypersensitivity reactions. Here we demonstrate that poly(lactic-co-glycolic acid) (PLGA) nanoparticles carrying rapamycin, but not free rapamycin, are capable of inducing durable immunological tolerance to co-administered proteins that is characterized by the induction of tolerogenic dendritic cells, an increase in regulatory T cells, a reduction in B cell activation and germinal centre formation, and the inhibition of antigen-specific hypersensitivity reactions. Intravenous co-administration of tolerogenic nanoparticles with pegylated uricase inhibited the formation of ADAs in mice and non-human primates and normalized serum uric acid levels in uricase-deficient mice. Similarly, the subcutaneous co-administration of nanoparticles with adalimumab resulted in the durable inhibition of ADAs, leading to normalized pharmacokinetics of the anti-TNFα antibody and protection against arthritis in TNFα transgenic mice. Adjunct therapy with tolerogenic nanoparticles represents a novel and broadly applicable approach to prevent the formation of ADAs against biologic therapies.
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Tolerancia Inmunológica/efectos de los fármacos , Nanopartículas/administración & dosificación , Sirolimus/administración & dosificación , Vacunas Sintéticas/inmunología , Adalimumab/administración & dosificación , Adalimumab/inmunología , Anafilaxia , Animales , Artritis Experimental/tratamiento farmacológico , Resorción Ósea/tratamiento farmacológico , Sistemas de Liberación de Medicamentos , Femenino , Hiperuricemia/tratamiento farmacológico , Ácido Láctico , Macaca fascicularis , Ratones Transgénicos , Nanopartículas/efectos adversos , Nanopartículas/química , Ácido Poliglicólico , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Ratas Sprague-Dawley , Sirolimus/inmunología , Linfocitos T Reguladores/efectos de los fármacos , Factor de Necrosis Tumoral alfa/genética , Vacunas Sintéticas/administración & dosificaciónRESUMEN
Disease management in vineyards with fungicides sometimes results in undesirable residue accumulations in grapes at harvest. Bioaugmentation of the grape fructosphere can be a useful approach for enhancing the degradation rate and reducing the residues to safe levels. This paper reports the in vitro and in vivo biodegradation of three triazole fungicides commonly used in Indian vineyards, by Bacillus strains, namely, DR-39, CS-126, TL-171, and TS-204, which were earlier found to enhance the dissipation rate of profenophos and carbendazim. The strains utilized the triazoles as carbon source and enhanced their in vitro rate of degradation. Myclobutanil, tetraconazole, and flusilazole were applied in separate vineyard plots at field doses of 0.40 g L(-1), 0.75 mL L(-1), and 0.125 mL L(-1), respectively. Residue analysis of field samples from the treated fields reflected 87.38 and >99% degradations of myclobutanil and tetraconazole, respectively, by the strain DR-39, and 90.82% degradation of flusilazole by the strain CS-126 after 15-20 days of treatment. In the respective controls, the corresponding percent degradations were 72.07, 58.88, and 54.28, respectively. These Bacillus strains could also simultaneously degrade the residues of profenofos, carbendazim, and tetraconazole on the grape berries and can be useful in multiclass pesticide residue biodegradation.
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Bacillus/metabolismo , Frutas/química , Fungicidas Industriales/metabolismo , Residuos de Plaguicidas/metabolismo , Triazoles/metabolismo , Vitis/química , Biodegradación Ambiental , Contaminación de Alimentos/prevención & control , Fungicidas Industriales/administración & dosificación , Especificidad de la Especie , Vitis/microbiologíaRESUMEN
Current treatments to control pathological or unwanted immune responses often use broadly immunosuppressive drugs. New approaches to induce antigen-specific immunological tolerance that control both cellular and humoral immune responses are desirable. Here we describe the use of synthetic, biodegradable nanoparticles carrying either protein or peptide antigens and a tolerogenic immunomodulator, rapamycin, to induce durable and antigen-specific immune tolerance, even in the presence of potent Toll-like receptor agonists. Treatment with tolerogenic nanoparticles results in the inhibition of CD4+ and CD8+ T-cell activation, an increase in regulatory cells, durable B-cell tolerance resistant to multiple immunogenic challenges, and the inhibition of antigen-specific hypersensitivity reactions, relapsing experimental autoimmune encephalomyelitis, and antibody responses against coagulation factor VIII in hemophilia A mice, even in animals previously sensitized to antigen. Only encapsulated rapamycin, not the free form, could induce immunological tolerance. Tolerogenic nanoparticle therapy represents a potential novel approach for the treatment of allergies, autoimmune diseases, and prevention of antidrug antibodies against biologic therapies.
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Antígenos/administración & dosificación , Antígenos/química , Tolerancia Inmunológica , Terapia de Inmunosupresión/métodos , Nanopartículas/química , Animales , Linfocitos T CD4-Positivos/inmunología , Modelos Animales de Enfermedad , Encefalomielitis Autoinmune Experimental/inmunología , Encefalomielitis Autoinmune Experimental/prevención & control , Encefalomielitis Autoinmune Experimental/terapia , Factor VIII/inmunología , Femenino , Hemocianinas/administración & dosificación , Hemofilia A/inmunología , Hemofilia A/terapia , Humanos , Hipersensibilidad Tardía/inmunología , Hipersensibilidad Tardía/terapia , Inmunidad Humoral , Inmunosupresores/administración & dosificación , Ácido Láctico/química , Ratones , Ratones Endogámicos BALB C , Nanocápsulas/administración & dosificación , Nanocápsulas/química , Nanopartículas/administración & dosificación , Oligodesoxirribonucleótidos/administración & dosificación , Ovalbúmina/administración & dosificación , Ovalbúmina/inmunología , Fragmentos de Péptidos/administración & dosificación , Fragmentos de Péptidos/inmunología , Péptidos/administración & dosificación , Péptidos/química , Péptidos/inmunología , Ácido Poliglicólico/química , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Proteínas/administración & dosificación , Proteínas/química , Proteínas/inmunología , Proteínas Recombinantes/inmunología , Sirolimus/administración & dosificaciónRESUMEN
Food safety is a global concern due to the increased use of pesticides in agriculture. In grapes, carbendazim is one of the frequently detected fungicides. However, it is amenable to biodegradation. In this study, we aimed to assess the degradation of carbendazim by four Bacillus subtilis strains, which had earlier shown potential for biocontrol of grape diseases. In liquid medium, each of the four strains, namely, DR-39, CS-126, TL-171, and TS-204, could utilize carbendazim as the sole carbon source. The half-life was minimized from 8.4 days in the uninoculated spiked control to 4.0-6.2 days by the four strains. In Thompson Seedless sprayed with carbendazim at 1.0 g L(-1), the residue on grape berries in control was 0.44 mg kg(-1) after 25 days of application, whereas in grapes treated with the four B. subtilis strains, the residues had decreased to 0.02 mg kg(-1). The degradation kinetics showed low half-lives of 3.1 to 5.2 days in treated grapes as compared to 8.8 days in control. In inoculated soils, the half-lives were 5.9 to 7.6 days in autoclaved and 6.5 to 7.2 days in nonautoclaved soils as compared to 8.2 and 8.0 days in respective controls. The growth dynamics of these strains in all the three matrices was not affected by presence of carbendazim. Bacillus strains TS-204 and TL-171 showed higher degradation rate than the other two strains in all the three matrices and show promise for in situ biodegradation of carbendazim.
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Bacillus subtilis/metabolismo , Bencimidazoles/metabolismo , Carbamatos/metabolismo , Fungicidas Industriales/metabolismo , Microbiología del Suelo , Agricultura , Bacillus subtilis/clasificación , Bencimidazoles/análisis , Biodegradación Ambiental , Carbamatos/análisis , Monitoreo del Ambiente , Fungicidas Industriales/análisis , Semivida , Cinética , Suelo/clasificación , Contaminantes del Suelo/análisis , Contaminantes del Suelo/metabolismoRESUMEN
The biodegradation of profenofos, an organophosphorus insecticide, by four Bacillus subtilis strains, namely, DR-39, CS-126, TL-171, and TS-204, isolated from grapevines or grape rhizosphere was studied in liquid culture, on grape berries, and in vineyard soil. Each of the four B. subtilis strains enhanced the degradation of profenofos in all three matrices. Degradation rate constants were best obtained by first + first-order kinetics module. In nutrient broth spiked with 5 µg/mL profenofos, inoculation with B. subtilis strains DR-39, CS-126, TL-171, and TS-204 reduced the half-life (DT50) of profenofos to 4.03, 3.57, 2.87, and 2.53 days, respectively, from the DT50 = 12.90 days observed in the uninoculated control. In Thompson Seedless grapes sprayed with profenofos at a field dose of 1250 mL ai/ha, the DT50 values were 1.07, 1.00, 2.13, and 2.20 days in grapes inoculated with B. subtilis strains DR-39, CS-126, TL-171, and TS-204, respectively, as compared to 2.20 days in uninoculated grapes. These four B. subtilis strains also enhanced the degradation of profenofos in autoclaved soil (DT50 = 5.93, 7.47, 6.00, and 4.37 days) and in nonautoclaved soil (DT50 = 0.87, 2.00, 2.07, and 2.43 days) amended with 5 µg/g profenofos from the half-lives of 17.37 and 14.37 days in respective uninoculated soils. Growth dynamic studies indicated that all four B. subtilis strains were able to establish and proliferate on berries and soil equally well in the presence or absence of profenofos. Degradation product 4-bromo-2-chlorophenol was identified by GC-MS. Strain DR-39 was most effective in the natural environments of grape and soil.
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Bacillus subtilis/metabolismo , Insecticidas/metabolismo , Organotiofosfatos/metabolismo , Vitis/microbiología , Bacillus subtilis/química , Bacillus subtilis/genética , Bacillus subtilis/aislamiento & purificación , Biodegradación Ambiental , Cromatografía de Gases y Espectrometría de Masas , Semivida , Insecticidas/química , Cinética , Organotiofosfatos/química , Contaminantes del Suelo/química , Contaminantes del Suelo/metabolismoRESUMEN
Recently we have reported anti-TB properties of a new class of conformationally-constrained indeno[2,1-c]quinolines, which are although considerably active (MIC 0.39-0.78 µg/mL) suffered from intense solubility problems. We thought of improving their bioavailability by prodrugs approach. Accordingly esters of the "Lead" indeno[2,1-c]quinolines 1, 15 and 27 derivatives were synthesized and their prodrug nature at the physiological pH were confirmed. Prodrugs were evaluated for their antimycobacterial activity against Mycobacterium tuberculosis H37Rv by MABA assay to show that they have 2- to 4-fold improved anti-TB activities, increased aqueous solubility and superior selectivity index over their respective parent compounds. MIC of these prodrugs was in the range of <0.20-6.0 µg/mL, and in general, no cytotoxicity was observed in VERO cells.
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Antibacterianos/síntesis química , Antibacterianos/farmacología , Profármacos/síntesis química , Profármacos/farmacología , Quinolinas/metabolismo , Aminoácidos/química , Antibacterianos/toxicidad , Línea Celular , Humanos , Hidrólisis , Interacciones Hidrofóbicas e Hidrofílicas , Pruebas de Sensibilidad Microbiana , Mycobacterium tuberculosis/efectos de los fármacos , Profármacos/metabolismo , Profármacos/toxicidad , Solubilidad , Agua/químicaRESUMEN
Studies have shown linkage between microsatellite markers from the chromosome 5q31-q33 region with asthma, atopy and total IgE levels in the Singapore Chinese population. However, subsequent case-control studies failed to show association between the polymorphisms in the candidate genes from this region and asthma or related phenotypes. In this study, we investigated 20 asthma candidate genes from this region for all possible informative polymorphisms within our population, linkage disequilibrium (LD) structure and tagging SNP transferability from HapMap populations. We re-sequenced these genes and identified 267 polymorphisms including 26 insertion-deletions, four microsatellite markers and 237 single nucleotide polymorphisms. The region contained 17 distinct LD blocks with the largest within the serine peptidase inhibitor kazal type 5 (SPINK5) gene spanning 23 kb. Of the 267 polymorphisms identified, 40% are represented in HapMap Han Chinese from Beijing and 29% in Han Chinese from Denver. 72% of the polymorphisms can be represented by tagged SNPs from the HapMap Beijing Han Chinese population and are highly correlated in terms of minor allele frequencies and LD structure. Our data suggest that although the HapMap Han Chinese population from Beijing is very similar to the Singapore Chinese population, this similarity is insufficient to account for up to 28% of the polymorphisms in the local population.
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Asma/etnología , Asma/genética , Cromosomas Humanos Par 5/genética , Desequilibrio de Ligamiento , Pueblo Asiatico/genética , Humanos , Polimorfismo Genético , Proteínas Inhibidoras de Proteinasas Secretoras/genética , Inhibidor de Serinpeptidasas Tipo Kazal-5 , Singapur/etnologíaRESUMEN
AIMS: Diabetic foot lesions are a major medical, social, and economic problem and are the leading cause of hospitalization for patients with diabetes, worldwide. ESBL-producing bacteria may not be detectable by routine disc diffusion susceptibility test, leading to inappropriate use of antibiotics and treatment failure. There is not much information on ESBL-producing organisms causing diabetic foot infection. An attempt was therefore made to study the ESBL-producing Escherichia coli and Klebsiella pneumoniae in diabetic foot patients with type 2 diabetes mellitus. MATERIALS AND METHODS: A total of 134 isolates of E. coli and K. pneumoniae were obtained from tissue, pus swab, and wound swab samples from diabetic foot ulcers submitted for routine microbiological analysis during the period January to December 2005 from patients with diabetic foot infections who had type 2 diabetes mellitus, attending S. L. Raheja Hospital. The above isolates were tested for antimicrobial susceptibility by disc diffusion technique according to clinical and laboratory standards institute (CLSI) guidelines. The screening for ESBL production was done by phenotypic confirmatory test using ceftazidime disc in the presence and absence of clavulanic acid as recommended by CLSI. RESULTS: Among the 134 isolates, 54 (40.29%) were E. coli and 80 (59.70%) were K. pneumoniae; among which, ESBL production was detected in 31 (23.13%) isolates. Of these 31, 15 (48.38%) were E. coli and 16 (51.61%) were K. pneumoniae. All the ESBL-producing isolates were found to be 100% sensitive to carbapenem (imipenem and meropenem). Mortality was found to be 3.22%, the cause of death being septicemia leading to multiple organ failure. CONCLUSIONS: The prevalence of ESBLs among members of Enterobacteriaceae constitutes a serious threat to the current beta-lactam therapy, leading to treatment failure and consequent escalation of costs. There is an urgent need to emphasize rational use of drugs to minimize the misuse of available antimicrobials.
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Pie Diabético/complicaciones , Pie Diabético/microbiología , Infecciones por Escherichia coli/microbiología , Escherichia coli/enzimología , Infecciones por Klebsiella/microbiología , Klebsiella pneumoniae/enzimología , beta-Lactamasas/biosíntesis , Antibacterianos/farmacología , Diabetes Mellitus Tipo 2/complicaciones , Escherichia coli/aislamiento & purificación , Infecciones por Escherichia coli/mortalidad , Femenino , Humanos , Infecciones por Klebsiella/mortalidad , Klebsiella pneumoniae/aislamiento & purificación , Masculino , Pruebas de Sensibilidad Microbiana , Sepsis/etiología , Sepsis/mortalidad , beta-Lactamas/farmacologíaRESUMEN
In seeking to find diagnostic biomarkers in proteomic spectra, two significant problems arise. First, not only is there noise in the measured intensity at each m/z value, but there is also noise in the measured m/z value itself. Second, the potential for overfitting is severe: it is easy to find features in the spectra that accurately discriminate disease states but have no biological meaning. We address these problems by developing and testing a series of steps for pre-processing proteomic spectra and extracting putatively meaningful features before presentation to feature selection and classification algorithms. These steps include an HMM-based latent spectrum extraction algorithm for fusing the information from multiple replicate spectra obtained from a single tissue sample, a simple algorithm for baseline correction based on a segmented convex hull, a peak identification and quantification algorithm, and a peak registration algorithm to align peaks from multiple tissue samples into common peak registers. We apply these steps to MALDI spectral data collected from normal and tumor lung tissue samples, and then compare the performance of feature selection with FDR followed by classification with an SVM, versus joint feature selection and classification with Bayesian sparse multinomial logistic regression (SMLR). The SMLR approach outperformed FDR+SVM, but both were effective in achieving good diagnostic accuracy with a small number of features. Some of the selected features have previously been investigated as clinical markers for lung cancer diagnosis; some of the remaining features are excellent candidates for further research.