Whole-genome sequence and mass spectrometry study of the snow blight fungus Phacidium infestans (Karsten) DSM 5139 growing at freezing temperatures.

Phacidium infestans (synonym Gremmenia infestans) is a significant pathogen that impacts Pinus species across the northern regions of Europe and Asia. This study introduces the genome sequence of P. infestans Karsten DSM 5139 (Phain), obtained through Pacbio technology. The assembly resulted in 44 contigs, with a total genome size of 36,805,277 bp and a Guanine-Cytosine content of 46.4%. Genome-mining revealed numerous putative biosynthetic gene clusters that code for virulence factors and fungal toxins. The presence of the enzyme pisatin demethylase was indicative of the potential of Phain to detoxify its environment from the terpenoid phytoalexins produced by its host as a defense mechanism. Proteomic analysis revealed the potential survival strategies of Phain under the snow, which included the production of antifreeze proteins, trehalose synthesis enzymes, desaturases, proteins related to elongation of very long-chain fatty acids, and stress protein responses. Study of protein GH11 endoxylanase expressed in Escherichia coli showed an acidic optimum pH (pH 5.0) and a low optimum temperature (45 °C), which is reflective of the living conditions of the fungus. Mass spectrometry analysis of the methanol extract of Phain, incubated at - 3 °C and 22 °C, revealed differences in the produced metabolites. Both genomic and mass spectrometry analyses showed the ability of Phain to adapt its metabolic processes and secretome to freezing temperatures through the production of osmoprotectant and cryoprotectant metabolites. This comprehensive exploration of Phain's genome sequence, proteome, and secretome not only advances our understanding of its unique adaptive mechanisms but also expands the possibilities of biotechnological applications.

Progress and prospects of C4 trait engineering in plants.

Incorporating C4 photosynthetic traits into C3 crops is a rational approach for sustaining future demands for crop productivity. Using classical plant breeding, engineering this complex trait is unlikely to achieve its target. Therefore, it is critical and timely to implement novel biotechnological crop improvement strategies to accomplish this goal. However, a fundamental understanding of C3 , C4 , and C3 -C4 intermediate metabolism is crucial for the targeted use of biotechnological tools. This review assesses recent progress towards engineering C4 photosynthetic traits in C3 crops. We also discuss lessons learned from successes and failures of recent genetic engineering attempts in C3 crops, highlighting the pros and cons of using rice as a model plant for short-, medium- and long-term goals of genetic engineering. This review provides an integrated approach towards engineering improved photosynthetic efficiency in C3 crops for sustaining food, fibre and fuel production around the globe.

Pharmacological potential of seaweed-associated heterotrophic Firmicutes.

Seaweed-associated bacterial symbionts are sources of potential pharmacological properties. The present study resulted in the culture-dependent isolation of bioactive heterotrophs belonging to the bacterial phylum Firmicutes, which were dominated more than 30% of the 127 cultivable isolates, amongst which 23 of them showed potential antimicrobial activities against a wide range of pathogens. The symbionts isolated from the seaweed Sargassum wightii showed significant bioactivity. Those were characterized as Bacillus safensis MTCC13040, B. valismortis MTCC13041, B. velezensis MTCC13044, B. methylotrophicus MTCC13042, Oceanobacillus profundus MTCC13045, B. tequilensis MTCC13043, and B. altitudinis MTCC13046. The organic extracts of the studied isolates showed potential antimicrobial properties against methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococci (minimum inhibitory concentration 6·25-12·5 µg ml-1 ). The organic extract of B. altitudinis MTCC13046 displayed significantly greater radical quenching ability (IC90 133 µg ml-1 , P < 0·05) other than attenuating hydroxymethyl glutaryl coenzyme A reductase (IC90 10·21 µg ml-1 , P < 0·05) and angiotensin-converting enzyme-1 (IC90 498 µg ml-1 , P < 0·05) relative to other studied heterotrophs. The organic extract of B. tequilensis MTCC13043 displayed significantly greater attenuation potential against pro-inflammatory 5-lipooxygenase (IC90 5·94 µg ml-1 , P < 0·05) and dipeptidyl peptidase-4 (IC90 271 µg ml-1 , P < 0·05). The seaweed-associated B. altitudinis MTCC13046 and B. tequilensis MTCC13043 could be used to develop promising pharmacological leads.

Relative contribution of different members of OsDREB gene family to osmotic stress tolerance in indica and japonica ecotypes of rice.

Drought/osmotic stress is the single largest production constraint in rain-fed rice cultivation. Different members of the DREB gene family are known to contribute to osmotic stress tolerance. In this study, an attempt was made to understand their relative contribution towards osmotic stress tolerance in indica and japonica ecotypes of rice. Two genotypes (one tolerant and one susceptible) from each ecotype were grown hydroponically, and 21-day-old seedlings were subjected to polyethylene glycol-induced osmotic stress (15% PEG-6000, equivalent to -3.0 bars osmotic potential). The tolerant genotypes CR143 and Moroberekan were found to have superior root traits (total root length, surface area and volume), better plant water status and increased total dry biomass as compared to their susceptible counterparts after 10 days of osmotic stress. Different members of the DREB gene family were differentially induced in response to osmotic shock (1 h after stress) and osmotic stress (24 h after stress), which also differed between the two rice ecotypes. From the gene expression profiles of 10 DREB genes (both DREB1 and DREB2 families), in indica two DREB genes, DREB1B and DREB1G, were significantly correlated with stress tolerance indices, whereas in japonica significant correlations with five DREB genes (DREB1A, DREB1B, DREB1D, DREB1E and DREB2B) were observed. We found that only one member, i.e. DREB1B, showed a significant correlation with drought tolerance indices in both indica and japonica ecotypes. This study provides an overview of the relative contribution of different members of the DREB gene family and their association with drought/osmotic stress tolerance in rice.

Combined application of ascorbic acid and endophytic N-fixing Azotobacter chroococcum Avi2 modulates photosynthetic efficacy, antioxidants and growth-promotion in rice under moisture deficit stress.

This group has previously reported the role of ascorbic acid (AA) as an antioxidant for survivability and ability to enhancing diazotrophic efficacy in Azotobacter chroococcum Avi2 under hydrogen peroxide (H2O2) stress. However, the present study showed the combined application of AA and Avi2 in drought-susceptible (IR64 and Naveen) and drought-tolerant (Ankit and Satyabhama) rice cultivars to determine their photosynthetic efficacy (chlorophyll fluorescence-imaging), antioxidants, and plant growth-promotion (PGP) under moisture deficit stress (MS, -60 kPa). The results indicated that combined application of AA and Avi2 significantly (p < 0.05) increased the total chlorophyll, relative water content, electrolytic leakage, super oxide dismutase, and catalase activities in all rice cultivars as compared to other MS treatments, whereas stress indicators like proline and H2O2 contents were proportionally increased under MS and their concentration were normalized under combined application of AA and Avi2. Photochemical quenching, non-photochemical quenching, photosynthetic electron transport rate, and the effective quantum efficiency were found to be increased significantly (p < 0.05) in Avi2 + AA as compared to other MS treatments. Moreover, rice roots harbored significantly (p < 0.05) higher copy number of nifH gene in Avi2 + AA treatment followed by Avi2 compared to flooded control and other MS treatments. Combined application of AA and Avi2 also increased the grain yield significantly (p < 0.05) by 7.09 % and 3.92 % in drought-tolerant (Ankit and Satyabhama, respectively) and 31.70 % and 34.19 % in drought-susceptible (IR64 and Naveen, respectively) rice cultivars compared to MS treatment. Overall, the present study indicated that AA along with Avi2 could be an effective formulation to alleviate MS vis à vis enhances PGP traits in rice.

Macrocyclic polyketides with siderophore mode of action from marine heterotrophic Shewanella algae: Prospective anti-infective leads attenuate drug-resistant pathogens.

AIMS: Biotechnological and chemical characterization of previously undescribed homologous siderophore-type macrocyclic polyketides from heterotrophic Shewanella algae Microbial Type Culture Collection (MTCC) 12715 affiliated with Rhodophycean macroalga Hypnea valentiae of marine origin, with significant anti-infective potential against drug-resistant pathogens. METHODS AND RESULTS: The heterotrophic bacterial strain in symbiotic association with intertidal macroalga H. valentiae was isolated to homogeneity in a culture-dependent method and screened for bioactivities by spot-over-lawn assay. The bacterial organic extract was purified and characterized by extensive chromatographic and spectroscopic methods, respectively, and was assessed for antibacterial activities with disc diffusion and microtube dilution methods. The macrocyclic polyketide compounds exhibited wide-spectrum of anti-infective potential against clinically significant vancomycin-resistant Enterococcus faecalis (VREfs), methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa and Klebsiella pneumonia with minimum inhibitory concentration of about 1-3 µg ml-1 , insomuch as the antibiotics chloramphenicol and ampicillin were active at ≥6·25 µg ml-1 . The studied compounds unveiled Fe3+ chelating activity, which designated that their prospective anti-infective activities against the pathogens could be due to their siderophore mechanism of action. In support of that, the bacterium exhibited siderophore production on bioassay involving the cast upon culture agar plate, and the presence of siderophore biosynthetic gene (≈1000 bp) (MF 981936) further corroborated the inference. In silico molecular modelling with penicillin-binding protein (PBP2a) coded by mecA genes of MRSA (docking score -11·68 to -12·69 kcal mol-1 ) verified their in vitro antibacterial activities. Putative biosynthetic pathway of macrocyclic polyketides through stepwise decarboxylative condensation initiated by malonate-acyl carrier protein further validated their structural and molecular attributes. CONCLUSIONS: The studied siderophore-type macrocyclic polyketides from S. algae MTCC 12715 with significant anti-infective potential could be considered as promising candidates for pharmaceutical and biotechnological applications, especially against emerging multidrug-resistant pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: This study exhibited the heterotrophic bacteria in association with intertidal macroalga as propitious biological resources to biosynthesize novel antibacterial agents.

Antibacterial aryl-crowned polyketide from Bacillus subtilis associated with seaweed Anthophycus longifolius.

AIMS: Microbiological, biotechnological and chemical characterization of a previously undescribed aryl-crowned polyketide from Bacillus subtilis MTCC 10403 isolated from brown seaweed Anthophycus longifolius with activity against opportunistic Gram-negative food-borne pathogenic bacterial strains. METHODS AND RESULTS: A culture-dependent method was used to isolate heterotrophic B. subtilis associated with A. longifolius and assessed for its antimicrobial properties. Minimum inhibitory concentration (MIC) of the title compound against the test pathogens was analysed by microtube dilution coupled with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide-based colorimetric endpoint detection. Bacillus subtilis MTCC 10403 was found to be antagonistic against Gram-negative food-borne pathogenic Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Salmonella enterica serotype Typhi, Aeromonas hydrophila and Vibrio sp. (diameter of zone of growth inhibition 13-22 mm). Bacillus subtilis was assessed for the presence of secondary metabolite coding polyketide synthase (pks) gene (KC589397, 700-bp gene product) and carboxylate siderophore framework in the aryl-crowned polyketide designated as 7-O-6'-(2″-acetylphenyl)-5'-hydroxyhexanoate-macrolactin by exhaustive spectroscopic techniques. The MIC assay showed that the reference antibiotics tetracycline and ampicillin were active at 25 µg ml-1 against the test pathogens, whereas the newly isolated polyketide displayed anti-infective properties against E. coli, A. hydrophilla, P. aeruginosa and Vibrio sp. at a lower concentration (MIC <13 µg ml-1 ). The MIC of the aryl macrolactin against K. pneumoniae was comparable with that of the referral antibiotics (~25 µg ml-1 ). The mode of antimicrobial action of acryl-crowned macrolactin was found to be iron chelating similar to siderophores. Putative biosynthetic pathway of the pks gene product further validated its molecular attributions. CONCLUSIONS: This study recognized new variant of antimicrobial aryl-crowned polyketide bearing methyl 6'-(2″-acetylphenyl)-5'-hydroxyhexanoate moiety at the C-7 position of the macrolactin system from A. longifolius-associated bacterium B. subtilis. SIGNIFICANCE AND IMPACT OF THE STUDY: This study revealed seaweed-associated micro-organisms as promising biological strata to produce new-generation anti-infective agents.

FAST implementation in Bangladesh: high frequency of unsuspected tuberculosis justifies challenges of scale-up.

SETTING: National Institute of Diseases of the Chest and Hospital, Dhaka; Bangladesh Institute of Research and Rehabilitation in Diabetes, Endocrine and Metabolic Disorders, Dhaka; and Chittagong Chest Disease Hospital, Chittagong, Bangladesh. OBJECTIVE: To present operational data and discuss the challenges of implementing FAST (Find cases Actively, Separate safely and Treat effectively) as a tuberculosis (TB) transmission control strategy. DESIGN: FAST was implemented sequentially at three hospitals. RESULTS: Using Xpert® MTB/RIF, 733/6028 (12.2%, 95%CI 11.4-13.0) patients were diagnosed with unsuspected TB. Patients with a history of TB who were admitted with other lung diseases had more than twice the odds of being diagnosed with unsuspected TB as those with no history of TB (OR 2.6, 95%CI 2.2-3.0, P < 0.001). Unsuspected multidrug-resistant TB (MDR-TB) was diagnosed in 89/1415 patients (6.3%, 95%CI 5.1-7.7). Patients with unsuspected TB had nearly five times the odds of being diagnosed with MDR-TB than those admitted with a known TB diagnosis (OR 4.9, 95%CI 3.1-7.6, P < 0.001). Implementation challenges include staff shortages, diagnostic failure, supply-chain issues and reliance on external funding. CONCLUSION: FAST implementation revealed a high frequency of unsuspected TB in hospitalized patients in Bangladesh. Patients with a previous history of TB have an increased risk of being diagnosed with unsuspected TB. Ensuring financial resources, stakeholder engagement and laboratory capacity are important for sustainability and scalability.

Antimicrobial properties of cultivable bacteria associated with seaweeds in the Gulf of Mannar on the southeast coast of India.

In this study, 234 bacterial strains were isolated from 7 seaweed species in the Gulf of Mannar on the southeast coast of India. The strains having consistent antimicrobial activity were chosen for further studies, and this constituted about 9.8% of the active strains isolated. Phylogenetic analysis using 16S rDNA sequencing with the help of classical biochemical identification indicated the existence of 2 major phyla, Firmicutes and Proteobacteria. Antimicrobial activity analysis combined with the results of amplifying genes encoding for polyketide synthetase and nonribosomal peptide synthetase showed that seaweed-associated bacteria had broad-spectrum antimicrobial activity. These epibionts might be beneficial to seaweeds by limiting or preventing the development of competing or fouling bacteria. Phylogenetic analysis of ketosynthase (KS) regions with respect to the diverse range of KS domains showed that the KS domains from the candidate isolates were of Type I. The bacterial cultures retained their antimicrobial activities after plasmid curing, which further suggested that the antimicrobial activity of these isolates was not encoded by plasmid, and the genes encoding the antimicrobial product might be present within the genome. Seaweed-associated bacteria with potential antimicrobial activity suggested that the seaweed species are an ideal ecological niche harboring specific bacterial diversity representing a largely underexplored source of antimicrobial secondary metabolites.

Successful treatment of nilotinib-induced pleural effusion with prednisone.

Chronic myeloid leukemia is characterized by a unique reciprocal translocation between chromosomes 9 and 22 resulting in deregulated tyrosine kinase activity. Tyrosine kinase inhibitors, such as imatinib, dasatinib, and nilotinib have revolutionized treatment of Chronic myeloid leukemia. However, tyrosine kinase inhibitors' use has presented new challenges in managing both acute and chronic toxicities, particularly 'off-target' toxicities like pleural effusion. Pleural effusions are seen less often with imatinib and very rarely with nilotinib. A 66-year-old male presented to emergency department with complaints of mild chest pain and dyspnea of 3 days duration with progressive worsening, including dyspnea at rest. Patient was currently taking nilotinib after failing imatinib for chronic myeloid leukemia. Nilotinib was put on hold. After exclusion of cardiac and pulmonary etiologies patient was treated for community acquired pneumonia with minimal improvement. Despite the very low incidence of pleural effusion with nilotinib (<1%), he was started on 20 mg of prednisone PO for 3 days. Patient had a dramatic improvement within 48 h after beginning prednisone. This treatment approach suggests that pleural effusions associated with nilotinib can be successfully treated in the same way as pleural effusions associated with dasatinib.

Morphometry of seminiferous tubules of human testes in different age groups in Bangladeshi cadavers.

This cross sectional descriptive study was done to observe the mean diameter of seminiferous tubules and to determine its differences between different age groups in Bangladeshi male. Thirty human testes of different age groups were collected by purposive sampling technique. Among them 22 specimens were collected from cadavers during routine postmortem examination and 8 specimens were from dead fetuses from Gynaecology & Obstetrics Department. The specimens were grouped into three categories Group A (28 to 42 weeks of gestational age), Group B (Up to 14 years) and Group C (15 to 70 years). The mean diameter of seminiferous tubules was measured and significant differences of the dimensions between different age groups were determined. The mean±SD diameter was 85.37±15.51 µm in Group A, 144.04±63.34 µm in Group B and 227.92±22.47 µm in Group C. Statistically, differences between age groups were calculated by using Unpaired Students 't' test. The present study revealed that the diameter increased with age and mean differences were statistically significant between Groups A&C, B&C and A&B.

Differential expression of salt overly sensitive pathway genes determines salinity stress tolerance in Brassica genotypes.

The objective of the present study was to examine the role of SOS pathway in salinity stress tolerance in Brassica spp. An experiment was conducted in pot culture with 4 Brassica genotypes, i.e., CS 52 and CS 54, Varuna and T 9 subjected to two levels of salinity treatments along with a control, viz., 1.65 (S(0)), 4.50 (S(1)) and 6.76 (S(2)) dS m(-1). Salinity treatment significantly decreased relative water content (RWC), membrane stability index (MSI) and chlorophyll (Chl) content in leaves and potassium (K) content in leaf, stem and root of all the genotypes. The decline in RWC, MSI, Chl and K content was significantly less in CS 52 and CS 54 as compared to Varuna and T 9. In contrast, the sodium (Na) content increased under salinity stress in all the plant parts in all the genotypes, however, the increase was less in CS 52 and CS 54, which also showed higher K/Na ratio, and thus more favourable cellular environment. Gene expression studies revealed the existence of a more efficient salt overly sensitive pathway composed of SOS1, SOS2, SOS3 and vacuolar Na(+)/H(+) antiporter in CS 52 and CS 54 compared to Varuna and T 9. Sequence analyses of partial cDNAs showed the conserved nature of these genes, and their intra and intergenic relatedness. It is thus concluded that existence of an efficient SOS pathway, resulting in higher K/Na ratio, could be one of the major factor determining salinity stress tolerance of Brassica juncea genotypes CS 52 and CS 54.

Mediastinal teratoma mimicking massive pleural effusion.

Immature mediastinal teratoma is very rare, found in only 1% of all mediastinal teratomas. Raised serum alpha feto-protein acts as important surrogate marker for both diagnosis and follow up in such cases. Surgery and adjuvant chemotherapy are keys in the management, especially in patients older than 15 years of age. We present a 14-year-boy presenting clinico-radiologically as left sided massive pleural effusion. Raised serum marker as well as excision biopsy of the mediastinal mass following thoracotomy were indicative of a diagnosis of immature teratoma.

Characterization of chemical modification of tryptophan by matrix-assisted laser desorption/ionization mass spectrometry.

Tert- butylation of tryptophan (2', 5', 7'- tri tertiary butyl tryptophan), formed during acidolytic cleavage of synthetic peptides Ac-KLVYWAE-CONH(2) (A-YW) and Ac-KLVWWAE-CONH(2) (A-WW), that are analogs of the fragment of Alzheimer's beta-amyloid peptide Ac-KLVFFAE-CONH(2), during solid-phase peptide synthesis, was characterized by matrix-assisted laser desorption/ionization time of flight/time of flight (MALDI TOF/TOF) mass spectrometry. Crude peptide was fractionated by high performance liquid chromatography. Peptide fractions were sequenced and modified tryptophan was determined with the help of MALDI TOF/TOF mass spectra. Thus, it is possible to pinpoint the particular tryptophan residue that undergoes modification during synthesis of peptides containing multiple tryptophan residues.

Internet addiction: consensus, controversies, and the way ahead.

OBJECTIVES: To review the fast-growing literature on Internet addiction. METHODS: Descriptive review, using electronic databases as well as hand-search of relevant publications or cross-references from 1970 to 2010. RESULTS: There are no universally accepted definitions for the captioned condition, but investigators seem to agree that it involves problematic computer usage that is time-consuming and causes distress or impairs functioning in important life domains. Several aetiological models have been proposed, from the diverse perspectives of learning theory, cognitive behavioural theory, social learning, reward deficiency, culture, genetics and neurobiology. Controversies abound, ranging from conceptual (whether behavioural addictions are true addictions), technical (which component of Internet use is a person 'addicted' to), and practical (how should Internet addiction be diagnosed, if it exists at all). However, using various instruments and populations, Internet addiction has been suggested as having a prevalence of 0.3 to 38%, with a young male preponderance. Several screening, diagnostic, and severity assessment instruments are now available, but few have been subjected to rigorous psychometric testing. Psychiatric co-morbidity is common. Treatment modalities lack a firm evidence base, but antidepressants, mood stabilisers, and cognitive behavioural therapy and other psychotherapies have been used. CONCLUSIONS: Recently, the American Psychiatric Association recommended including Internet addiction in its forthcoming 5th edition of the Diagnostic and Statistical Manual of Mental Disorders, but only as an appendix and not in the main body of the addictive disorders. This appears to be a fairly balanced and cautious approach, which can hopefully give rise to more meaningful research in this important but controversial area.

Restoration by aspirin of impaired plasma maspin level in human breast cancer.

Maspin, an anti breast cancer protein, is produced in the normal mammary cells but not in malignant cells in breast cancer. We investigated the effect of aspirin induced increase of plasma nitric oxide (NO) on plasma maspin production in breast cancer patients. Fifteen breast cancer patients (35-65 years), who had not yet undergone any cancer therapy, and an equal number of age matched normal female volunteers participated in the study. They were asked not to take any medication for two weeks. All participants then ingested 150 mg of aspirin. Plasma NO and maspin levels were determined before and at 60 min after the ingestion of aspirin. It was found that the maspin level in plasma increased to 4.63+/-0.02 nM from the basal 0.95+/-0.012 nM (p<0.001) with increase of plasma NO from 0.60+/-0.03 microM to 2.08+/-0.030 microM (p<0.001) in breast cancer patients. In normal volunteers the basal maspin increased from 4.76+/-0.041 to 9.36+/-0.036 nM (p<0.001) with increase of NO from 2.15+/-0.08 to 3.36+/-0.04 microM (p<0.001) at the same period. These results indicated that the ingestion of aspirin might be beneficial for breast cancer through increased maspin production.

An unusual presentation of medial sphenoid wing meningioma.

A young woman of 26 years attended eye OPD with the complaints of complete and rapid loss of vision of her righ eye and progressive dimness of vision of left eye, frontal headache and nausea for last two months. On eye examination, she was found to have no perception of light to her right eye. Ophthalmoscopy examination revealed primary optic atrophy in right eye and marked papilloedema to her left eye. CT scan of brain showed space occupying lesion in right parasellar region with sellar and suprasellar extension with involvement of right anterior clinoid process suggesting medial sphenoid wing meningioma. The patient underwent surgery and the histopathology proved the diagnosis as meningioma. The case of medial sphenoid wing meningioma in a young female producing rapid Foster Kennedy syndrome is reported for its unusual presentation.

Therapeutic and hemolytic evaluation of in-situ liposomal preparation containing amphotericin - beta complexed with different chemically modified beta - cyclodextrins.

PURPOSE: The objective of this study was to evaluate therapeutic and haemolytic effects of liposomal preparation derived from proliposome entrapping inclusion complex of amphotericin B (AmB) with the chemically modified beta-cyclodextrin (beta-CD). METHODS: a series of liposomal AmB formulations with varying beta-CD i.e. Hydroxy propyl beta-CD (HPBCD) and Sulfo butyl ether beta-CD (SBEBCD) having similar AmB content (0.5 mg/kg) were prepared and their effect compared with conventional liposomal amphotericin B (L-AmB) and free AmB on erythrocyte lysis and antifungals activity in experimental aspergillosis- and Cryptococcosis- mice model in-vivo. RESULTS: the liposomal AmB - HPBCD and AmB - SBEBCD found to be 6 times less toxic than free AmB or conventional liposomal AmB. Experimental findings indicate that infected animals treated with L-AmB entrapped inclusion complexes significantly reduced CFU values (fungal counts), whereas infected animals treated with conventional liposome or free AmB showed insignificant reduction in CFU. A marked increase in the percent survival was observed in the case of animals treated with liposomal AmB formulation (HPBCD/SBEBCD). Furthermore, the in-vitro toxicity (haemolysis) of the proliposome-based liposomal vesicles (PBLV) entrapped AmB-SBEBCD/HPBCD at 37 degrees C was approx. 50% at maximum of the conventional liposomal AmB at a dose of 118 microg/ml as measured after 1 hr. incubation. CONCLUSIONS: the results of these experiments permitted us to conclude that the stabilization of liposome derived from proliposome entrapping inclusion complex of amphotericin B (AmB) with beta-CD could serve an alternative approach to enhance the therapeutic window of AmB in clinical medicine.

Massive pancreatico-pleural effusion--an often unrecognised entity.

Massive haemorrhagic pleural effusion secondary to pancreatitis in a five year old girl is described. The diagnosis was established on the basis of an exudative pleural effusion with an amylase level above 4,000 IU/l. Management of effusion was by intercostal tube drainage and antibiotics.

PHARMACOKINETIC STUDIES OF IN-SITU LIPOSOMAL PREPARATION CONTAINING AMPHOTERICIN B COMPLEXED WITH DIFFERENT CHEMICALLY MODIFIED beta-CYCLODEXTRINS.

The purpose of this study was to evaluate the influence of chemically modified beta-cyclodextrin (beta-CD) which could affect the in-vivo stabilization of liposomal preparation derived from proliposome entrapping inclusion complex of amphotericin B (AmB) with beta-CD. A series of liposomal AmB formulations with varying beta-CD i.e. Hydroxypropyl beta-CD (HPBCD) & Sulfo butyl ether beta-CD (SBEBCD) and lipid dose having similar AmB content (0.5 mg/kg) were compared with conventional liposomal amphotericin B (L-AmB) or free AmB in rats and their pharmacokinetic data were analyzed considering the varying volume of distribution with respect to the varying lipid concentration (65mg to 110mg) in blood. These results indicate that L-AmB entrapped inclusion complexes safely achieved higher Cmax (P < 0.05) & AUC (P < 0.02) and demonstrated saturable, nonlinear elimination from plasma via reticuloendothelial organ uptake at higher lipid level (>80mg) as compared with conventional L-AmB or free AmB. Furthermore in-vivo stabilization potential for liposomal preparation via AmB/ beta-Cyclodextrin inclusion complexes appeared to be in pattern of HPBCD < SBEBCD. It is concluded that the preparation of liposome derived from proliposome entrapping inclusion complex of amphotericinB (AmB) with beta-CD could serve an alternative approach to enhance the therapeutic window of AmB in clinical medicine.