Effect on the Prevalence of Various Diffuse Parenchymal Lung Diseases due to Paradigm Change in the Guidelines

Background: Diffuse parenchymal lung diseases (DPLDs) have gone through various changes in nomenclature and classification since they were first described in 1868. Increasing knowledge about their etiopathogenesis has since led to several reclassifications and changes in the nomenclature. This has had a major impact on the prevalence of each interstitial lung disease (ILD) reported by the different registries worldwide. In this study, we attempted to describe the distribution of the different DPLDs in our population and reported changes in prevalence due to changing diagnostic criteria for the disease. Materials and methods: We analyzed retrospective data of 434 patients. For the initial 75 patients, ATS/ERS guidelines published in 2002 were followed in the diagnosis of the ILD (group I). In the later part of the study (359 patients), the diagnosis was based on the computed tomography (CT) patterns defined by ATS/ERS/JPS/ALAT statement on diagnosis of idiopathic pulmonary fibrosis (IPF) and updated 2013 ATS/ERS guidelines (group II). Results: Of the 75 patients in group I, IPF was the most common diagnosis (52%) made at that time, followed by sarcoidosis and connective tissue-related ILD (CTD-ILD) with 12% each. Group II had 359 patients, with IPF again being the most commonly diagnosed ILD with 21.3%. This was followed by CTD-ILD (18.6%), sarcoid (14.7%), and idiopathic nonspecific interstitial pneumonitis (iNSIP; 13.3%). The changing guidelines have an impact on reporting of different DPLD by our multidisciplinary teamover a period of time. Though IPF was the most commonest DPLD reported among both the groups, the diagnosis of IPF had fallen by more than half in the second group. It was paralleled by an increase in the diagnosis of iNSIP and chronic hypersensitivity pneumonitis. These reported changes in the prevalence of DPLDs may reflect the better-defined criteria in the latest guidelines and a better understanding of the fibrotic ILDs other than IPF by the multidisciplinary team. Conclusions: The frequency of diagnosis of the different DPLDs has changed, following the publication of several guidelines in the last decade. It has recognized newer entities with greater clarity, such as idiopathic NSIP and interstitial pneumonia with autoimmune features.

Pattern of drug-resistance and treatment outcome in multidrug-resistant pulmonary tuberculosis.

Aims and Objectives. To study the pattern of drug-resistance and treatment outcomes among patients with confirmed multidrug-resistant pulmonary tuberculosis (MDR-PTB). Methods. A prospective study was conducted at Rajiv Gandhi Institute of Chest Diseases, Bengaluru, Karnataka, India. Between January 2005 and December 2008, 224 confirmed MDR-PTB cases were studied for various drug-resistance patterns, and their treatment outcomes were analysed until November 2010. Sputum culture and drug sensitivity tests (DST) were carried out at National Tuberculosis Institute, Bengaluru; DST was done for all first-line drugs except pyrazinamide. Results. Of the 224 MDR-PTB patients, 146 (65.2%) were resistant to all first-line drugs, 39 (17.4%) to isoniazid, rifampicin and streptomycin; 19 (8.5%) to isoniazid, rifampicin and ethambutol; and 20 (8.9%) to isoniazid and rifampicin. Among them, 145 (64.7%) patients were cured, 5 (2.2%) had treatment-failure, 10 (4.4%) died, and 64 (28.5%) defaulted. Among 145 cured cases, 100 (69%) were resistant to all first-line drugs, 23 (16%) to isoniazid, rifampicin and streptomycin, 11(8%) to isoniazid, rifampicin and ethambutol, and 11(8%) to isoniazid and rifampicin. Conclusions. The most common pattern observed in this study was resistance to all four first-line drugs followed by resistance to isoniazid, rifampicin and streptomycin. Patients resistant to all first-line drugs had early sputum culture conversion and better cure rate as compared to other resistance patterns.

Comparative Study of Peripheral Blood Smear and Quantitative Buffy Coat in Malaria Diagnosis.

A rapid test for diagnosis of malaria based on acridine orange staining of centrifuged blood samples in a microhaematocrit tube (QBC) was compared with Leishman stained thin peripheral blood smear in 287 samples. Malaria was diagnosed in 44 patients by Leishman staining technique and in 65 patients by QBC method. The QBC method allowed detection of an additional 21 cases. Thus the prevalence rate of malaria during the study was 22.65%. In 222 Patients who were negative by the QBC technique, the Leishman stained smears were also negative for malarial parasite. Although QBC method was superior to the smear for malarial parasite detection, species identification was difficult by this technique. The QBC method provides a reliable, quick, easily mastered, accurate method for diagnosis of malaria. The QBC system can also be used in the diagnosis of other parasitic diseases from blood (Filariasis). However, Leishman stained thin blood film still appear superior for species identification.

Studies on carbohydrate moieties of the glycoprotein, glucoamylase II of Aspergillus niger: Nature of carbohydratepeptide linkage and structure of oligosaccharides.

Electrophoretically homogeneous type 1 (GP-C1, and GP-C 2), type 2 (GP-C3a and GP-C3b,) and type 3 (GP-D1, and GP-D2) glycopeptides from Aspergillus niger glucoamylase II (Manjunath and Raghavendra Rao, preceding paper) were separately treated with alkaline borohydride. The (ß-eliminated oligosaccharides were subjected to single and sequential digestion with specific glycosidases and the products analysed by gas liquid chromatography. The studies revealed that carbohydrate moieties were present as mannose, Man-Man-, and trisaccharide structures, namely, (a) GIc-Man-Man-, (b) Gal-Man-Man, (c) Man-Man-Man-, (d) GlcNAc-Man-Man-, and (e) Xyl-Man-Man. None of the glycopeptides contained all the trisaccharide structures (a) to (e). Type 1 glycopeptide contained structures (a), (b) and (c); type 2, (a) and (d)and type 3, (a), (b)and (e). The number of carbohydrate units (mono-, di-and trisaccharides) present in the major glycopeptides was determined and tentative structures for the glycopeptides proposed. Carbohydrate units appeared to occur in clusters of 4 to 7 in each glycopeptide, a structure unique to the carbohydrate moiety in Aspergillus niger glucoamylase. Based on carbohydrate analysis and yields of glycopeptide, the number of units of each type of glycopeptide present in glucoamylase II was tentatively calculated to give two of type Man:Glc:Gal = 12-15:l:l, one of type Man:Glc:GlcN = 10-l 1:1:2 and one of type Man :GIc :Gal :Xyl =4-8:0.1:0.5-0.8:0.3-1 glycopeptides.

Studies on carbohydrate moieties of Aspergillus niger glucoamylase II: lsolation,purification and characterization of glycopeptides.

Six glycopeptide fractions namely GP-C1, GP-C2. GP-C3a .GP-C3b.GP-D, and GPD2 were isolated after exhaustive digestion of glucoamylase II (Glucozyme) from Aspergillus niger with pronase. They were purified using gel-filtration. high-voltage paper electrophoresis and ion-exchange chromatography on Dowex-50 and Dowex-1. They appeared homogeneous on electrophoresis under different conditions of pHs. The molecular weights ranged from 1600 and 4000 for these glycopeptides. Ally of them contained serine at the N-terminal end. Serine and threonine were the major amino acids with glycine, alanine, proline and tryosine present as minor constituents. Carbohydrate analysis revealed the presence of different sugars. Based on this, the glycopeptides were grouped into three types: (1) GP-C1 and GP-C2 containing mannose, glucose and galactose; (2) GP-C3a, and GP-C3b,containing mannose glucose and glucosamine; and (3) GP-D1 and GP-D2, containing mannose. glucose, galactose and xylose. Most sugar constituents in each glycopeptide occured in non-integral ratios implying a microheterogeneity of the carbohydrate moiety in Aspergillus niger glucoamylase II·.

Immunochemical relationship between glucoamylases I and II of Aspergillus niger.

Rabbit antisera were prepared against the purified glucoamylases I and II of Aspergillus niger. Relationships between the two enzyme forms were investigated by using the antisera in immunodiffusion and immunoinhibition experiments. Both the forms of glucoamylase gave a single continuous precipitin band demonstrating very close structural resemblance. They gave almost identical immunoprecipitation patterns and had the same equivalence points indicating that the two forms of A. niger gluoamylases were immunologically identical. The enzyme treated with periodate was immunologically identical with the controls and had slightly less enzyme activity but showed greatly reduced stability on storage at 4° C.

Comparative studies on glucoamylases from three fungal sources.

Five commercial preparations of glucoamylases (three from Aspergillus niger, one each from Aspergillus foetidus and Aspergillus candidus) were purified by ultrafiltration, Sepharose gel filtration and DEAE sephadex chromatography. Two forms of the enzyme, namely glucoamylase I and glucoamylase II were obtained from the fungi except from one strain of A. niger. All the enzymes appeared homogeneous by electrophoresis and ultracentrifugation. The specific activities varied between 85 and 142 units. The pH and temperature optima were between 4 and 5, and 60° C respectively. The molecular weight as determined by the sodium dodecyl sulphate polyacrylamide gel electrophoresis ranged from 75,000 to 79,000 for glucoamylase I and 60,000 to 72,000 for glucoamylase II. Only A. niger gluco amylases contained phenylalanine at the N terminal end. The amino acid compo sition of the enzymes was generally similar. However, A. niger and A. foetidus glucoamylases, in contrast to A. candidus enzymes, contained greater percentage of acidic than of basic amino acids. The enzymes contained 15 to 30% carbohydrate and 49 to 57 residues of monosaccharides per mol. A. niger enzymes contained mannose, glucose, galactose, xylose and glucosamine but the A. candidus enzyme lacked xylose and glucose and only xylose was absent in A, foetidus enzymes.. Majo rity of the carbohydrate moieties were O glycosidically linked through mannose to the hydroxyl groups of seline and threonine of the polypeptide chain.