Potential use of microwave technology in dermatology.

BACKGROUND: Microwaves are used in medicine for diagnostics, and treatment of cancer. Recently, novel microwave devices (Swift®, Emblation Ltd, UK and miraDry®, Miramar Labs Inc., CA) have been cleared by the FDA and Health Canada for various dermatological conditions. OBJECTIVE AND METHODS: To review the dermatological use of microwave-based treatments (plantar warts, corns, actinic keratosis, dermatophytosis, axillary hyperhidrosis, osmidrosis, and hidradenitis suppurativa). Clinical trials, case reports, or in vitro studies for each condition are summarized. RESULTS AND CONCLUSION: Microwaves are a promising alternative therapy for cutaneous warts, actinic keratosis, axillary hyperhidrosis, and osmidrosis, with favorable safety profiles. However, patients with hidradenitis suppurativa have had negative clinical outcomes. Limited treatment of corns showed good pain reduction but did not resolve hyperkeratosis. A preliminary in vitro study indicated that microwave treatment inhibits the growth of T. rubrum. We present the first case of toenail onychomycosis successfully treated with microwaves. Despite the advancements in the use of microwaves, the mechanism of action in non-ablative treatment is not well understood; further research is needed. More high-quality randomized clinical trials with larger groups and long follow-up periods are also required to evaluate the clinical benefits and possible adverse effects of microwaves in treating dermatological conditions.

Characterisation of biomass resources in Nepal and assessment of potential for increased charcoal production.

Characterisation of 27 types of biomass was performed together with an assessment of regional resource availability. Charcoal was produced under two conditions from all samples and their yields were compared. Sugarcane bagasse, sal and pine produced the best charcoal with a low volatile matter and high calorific value. The amount of high-quality charcoal which can be made within Nepal from the biomass types tested is equivalent to 8,073,000 tonnes of firewood a year or 51% of the yearly demand. The areas which would benefit the most from charcoal making facilities are the Mid-hills of the Western, Central and Eastern Development Regions, as well as the Terai in the Central and Eastern Development Regions. The main potential benefit is to convert agricultural residues which are underutilised because, in their original form, produce large quantities of smoke, to cleaner burning charcoal. The conversion of agricultural residues to charcoal is also a viable alternative to anaerobic digestion in the Mid-hills.

Bovine glycomacropeptide promotes the growth of Bifidobacterium longum ssp. infantis and modulates its gene expression.

Bovine milk glycomacropeptide (GMP) is derived from κ-casein, with exclusively o-linked glycosylation. Glycomacropeptide promoted the growth of Bifidobacterium longum ssp. infantis in a concentration-dependent manner, and this activity was lost following periodate treatment of the GMP (GMP-P), which disables biological recognition of the conjugated oligosaccharides. Transcriptional analysis of B. longum ssp. infantis following exposure to GMP revealed a substantial response to GMP relative to bacteria treated with GMP-P, with a greater number of differentially expressed transcripts and larger fold changes versus the control. Therefore, stimulation of B. longum ssp. infantis growth by GMP is intrinsically linked to the peptide's O-linked glycosylation. The pool of differentially expressed transcripts included 2 glycoside hydrolase (family 25) genes, which were substantially upregulated following exposure to GMP, but not GMP-P. These GH25 genes were present in duplicated genomic islands that also contained genes encoding fibronectin type III binding domain proteins and numerous phage-related proteins, all of which were also upregulated. Homologs of this genomic arrangement were present in other Bifidobacterium species, which suggest it may be a conserved domain for the utilization of glycosylated peptides. This study provides insights into the molecular basis for the prebiotic effect of bovine milk GMP on B. longum ssp. infantis.

Menopausal Health Status of Nepalese Women.

INTRODUCTION: Menopause is an unavoidable and sometimes problematic condition in which women may suffer from a number of health problems. Knowledge of the major symptoms associated with menopause helps reduce the burden and stress associated with the condition. In this context, a study was conducted to establish baseline data regarding menopausal health problems among Nepalese women. METHODS: A descriptive cross-sectional study was conducted among 2000 women aged over 40 years to identify their knowledge, attitude and practice related to menopausal health problems. Data was collected by interviewing the women and doing physical examination. Analysis was done using descriptive statistics and SPSS software was used for data analysis. RESULTS: A cohort of 2073 women (ages 40-60 years) participated in the research, among them 2000 yielded complete response. The study revealed that 820 (41%) women had reached menopause. The average age of menopause was 48.7 years. Majority of the women 1183 (59.2%) in the study did not know about menopausal health problems. Abnormal bleeding 353 (17.65%), sweating 315 (15.75%), hot flushes 299 (14.95%), joint/muscle pain 285 (14.25%) were the most common menopausal symptoms known by the women. Joint pain 736 (36.8%), hot flushes 584 (29.2%), irregular bleeding 582 (29.1%) were the most common experienced symptoms of menopause and only 586 (29.3%) consulted the health workers to alleviate menopausal symptoms. Moreover, most of the women 926 (46.3%) accepted menopause as a part of life. CONCLUSIONS: The research has tried to establish the baseline of menopausal health problems in the Nepalese women. The average age of menopause was 48.7 years. Majority did not know about menopausal health problems. Further researches with nationally representative sample are recommended to further explore the menopausal health problems of Nepalese women.

The effect of hospital biocide sodium dichloroisocyanurate on the viability and properties of Clostridium difficile spores.

Clostridium difficile is the primary cause of healthcare-associated diarrhoea globally and produces spores which are resistant to commonly used biocides and are able persist on contaminated surfaces for months. This study examined the effect of sublethal concentrations of the biocide sodium dichloroisocyanurate (NaDCC) on the viability of spores produced by 21 clinical isolates of C. difficile representing a range of PCR ribotypes. Spores exposed to 500 ppm NaDCC for 10 min exhibited between a 4-6 log10 reduction in viability which was independent of spore PCR ribotype. The effect of sublethal concentrations of biocide on the surface properties of exosporium positive and negative clinical isolates was determined using a spore adhesion to hydrocarbon (SATH) assay. These isolates differed markedly in their responses suggesting that exposure to biocide can have a profound effect on hydrophobicity and thus the ability of spores to adhere to surfaces. This raises the intriguing possibility that sublethal exposure to NaDCC could inadvertently promote the spread of the pathogen in healthcare facilities. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first to report on changes in Clostridium difficile spore surface property after exposure to sublethal levels of the commonly used biocide sodium dichloroisocyanurate. The implications of these changes to the spore surface include increased adherence of the spores to inorganic surfaces which can directly contribute to persistence and spread of spores within the hospital environment.

Unique glycosignature for intervertebral disc and articular cartilage cells and tissues in immaturity and maturity.

In this study, on/off markers for intervertebral disc (IVD) and articular cartilage (AC) cells (chondrocytes) and distinct glycoprofiles of cell and tissue-types were identified from immaturity to maturity. Three and eleven month-old ovine IVD and AC tissues were histochemically profiled with a panel of lectins and antibodies. Relationships between tissue and cell types were analysed by hierarchical clustering. Chondroitin sulfate (CS) composition of annulus fibrosus (AF), nucleus pulposus (NP) and AC tissues was determined by HPLC analysis. Clear on/off cell type markers were identified, which enabled the discrimination of chondrocytes, AF and NP cells. AF and NP cells were distinguishable using MAA, SNA-I, SBA and WFA lectins, which bound to both NP cells and chondrocytes but not AF cells. Chondrocytes were distinguished from NP and AF cells with a specific binding of LTA and PNA lectins to chondrocytes. Each tissue showed a unique CS composition with a distinct switch in sulfation pattern in AF and NP tissues upon disc maturity while cartilage maintained the same sulfation pattern over time. In conclusion, distinct glycoprofiles for cell and tissue-types across age groups were identified in addition to altered CS composition and sulfation patterns for tissue types upon maturity.

Male Participation in Family Planning: Human Behaviour Perspective.

BACKGROUND: Family planning (FP) is one of the major components of reproductive health and its goal is to prevent unwanted pregnancies and regulate wanted pregnancies, thereby ensuring the health of mothers and children. It also aims at regulating the population in order to maintain the vital balance between development and the environment. Ideally, FP depends on the efforts of a couple where the man and woman are equally responsible and accountable. In reality, however, this is not the case. It is in this background that the present study aims at examining the nature and level of male participation in preventing unwanted pregnancies and the factors that influence male participation in FP. METHODS: The data for the study was derived using mixed methods, drawing from both quantitative and qualitative approaches. The research design was cross-sectional, descriptive and observational. RESULTS: Despite the high level of knowledge and awareness about FP among the respondents, it was found that male participation in FP continues to remain very low. The multidimensional factors influencing their participation include education of the couple (the unadjusted odds ratio of higher education level of respondents is 2.182 and the adjusted OR is 1.972; and the unadjusted OR of higher education level of the spouse is 2.030, and the adjusted OR is 1.570), and economic condition (the unadjusted OR of higher income is 2.272 and the adjusted OR is 2.436) of family. Male participation in FP was found to be higher in rural areas (69.8 %) and among Dalits (72.5%). Other factors include social stigma and religious practice, patriarchal notions, gender roles and individual characteristics. CONCLUSIONS: Male participation plays a crucial role in population management, but patriarchal notions, socioculturally defined gender roles, combined with the inefficiency of the current FP programme, and biological factors contribute towards keeping male participation very low.

Association of cytokine gene polymorphisms in patients with tuberculosis and their household contacts.

Cytokine gene polymorphisms are known to be associated with functional differences in cytokine regulation and may affect host susceptibility to tuberculosis (TB). Contacts are important group in developing tuberculosis infection and are 10-60 times more likely to develop TB than general population. The present study was carried out in patients with TB (N = 176), their household contacts (HHC; N = 155) from Free Chest TB Clinic PPM DOTS (1TU) covering 500,000 population at Mahavir Hospital and Research Centre, Hyderabad, and healthy controls (HC; N = 170) also included. The association of single-nucleotide polymorphisms (SNPs) in the promoter region of TNF-α (-308G/A), IL-2 (-330T/G), IL-4 (-589C/T) and in exon region of TGF-ß1 (+869T/C) genes was assessed by ARMS & PCR-RFLP using specific primers in the above-mentioned subjects. The differences in allelic or genotypic frequencies of TNF-α (-308G/A) between patients, their HHC and HC were not statistically significant (P > 0.05). IL-2 (-330T/G) TG genotype was significantly different between patients, HHC compared to HC (P < 0.002, OR-1.997, 95%CI-1.260-3.168, P < 0.03, OR-1.602, 955CI-1.003-2.561).IL-4 (-589C/T) CC genotype was significantly different between patients and HC (P < 0.03, OR-1.791, 95%CI-1.009-3.189) as well as between HHC and HC at P < 0.0001, OR-2.56, 95%CI-1.448-4.545. In addition, the TGF-ß 1 (+869T/C) TC genotype was significantly associated with susceptibility to tuberculosis in patients when compared against HC(P < 0.0001, OR-3.416, 95%CI-2.063-5.670) and HHC (P < 0.0001, OR-2.357, 95%CI-1.439-3.868), respectively.MDR analysis indicated that TT genotype of TGF-ß1 with TT and CT genotypes of IL-4 showed high risk with GA, TT genotypes of TNF-α, IL-2, respectively. Our results suggest that IL-2 (-330T/G), IL-4 (-589 C/T) and TGF-ß1 (+869T/C) gene polymorphisms may be associated with TB susceptibility.

Serum vitamin D levels and VDR polymorphisms (BsmI and FokI) in patients and their household contacts susceptible to tuberculosis.

Vitamin D, an immunomodulator of macrophage function, can activate human antimycobacterial activity. Vitamin D deficiency (VDD) is associated with an impaired mycobacterial immunity and susceptibility to tuberculosis. It has been found that vitamin D and its receptor may be essential for immune function. In this study, we examined the serum 25(OH) vitamin D levels and its receptor (VDR) polymorphisms with susceptibility to tuberculosis in patients, household contacts and healthy controls. Serum 25(OH) vitamin D levels were measured in 75 cases (25 patients, 25 household contacts and 25 healthy controls), and polymorphisms (BsmI and FokI) were carried out in 335 cases (110 patients, 110 household contacts and 115 healthy controls). The proportion of serum 25(OH) vitamin D deficiency and insufficiency was high in patients (44, 58%) and household contacts (40, 48%) compared to controls (48%). The BB and Bb genotypes of BsmI were significantly associated in patients (P < 0.014; OR: 0.509; CI: 0.265-0.876) (P < 0.001; OR: 2.351; CI: 1.368-4.041) and household contacts (P < 0.04; OR: 0.575; CI: 0.336-0.985); (P < 0.002; OR: -2.267; CI: 1.32-3.895) when compared to healthy controls. The diplotype and MDR analysis showed the high-risk genotypes of BsmI and FokI polymorphisms. Vitamin D deficiency and its association with VDR gene polymorphisms may be useful to identify the high-risk group individuals.

Carotid Doppler and lipid profile findings in ischemic stroke patients--a hospital based study.

A prospective study was carried out during 2010 - 2011 in the Department of Radiology & Imaging of Nepal Medical College and Teaching Hospital, Jorpati, Kathmandu and Om Hospital and Research Centre, Chabahil, Kathmandu in which a total of 45 patients with clinically and CT scan diagnosed ischaemic cerebrovascular accident (CVA) underwent carotid Doppler study. During the hospital stay these patients also underwent lipid profile estimation to see for presence or absence of hyperlipidemia. Of these 45 patients, CT scan showed left middle cerebral artery (MCA) infarctions in 12, right MCA infarction in 12, lacunar infarctions in 10, watershed territory infarction in 4, posterior circulation infarction in 1 and venous infarction in 1 patient. Carotid Doppler study showed more than 50% stenosis in 31 patients. Among these 31 patients, 28 had hyperlipidemia where as blood cholesterol and triglyceride values in 3 patients were normal. The present study like several other studies done in the past showed association between carotid stenosis and hyperlipidemia, leading to CVA.

Performance in the Duke-Elder ophthalmology undergraduate prize examination and future careers in ophthalmology.

AIMS: Cognitive factors (eg, academic achievement) have had a significant role in selecting postgraduate surgical trainees in the past. This project sought to determine the role of a national undergraduate ophthalmology prize examination (Duke-Elder examination) in the selection of postgraduate ophthalmology trainees. This would also serve as a quality assurance exercise for the assessment, in which the ultimate aim is to encourage trainees into ophthalmology. METHODS: A retrospective analysis of the top 20 ranked candidates in the Duke-Elder examination from 1989 to 2005 (except 1995) was carried out to determine which of them subsequently entered the ophthalmic training and General Medical Council Specialist Registers. RESULTS: Out of the top 20 candidates in the exam, 29.5% went into specialist training in ophthalmology. Some appeared in the top 20 more than once, with 56% of them going into ophthalmic training, but they had a similar median time to enter training as those who appeared in the top 20 once. There was no significant evidence to suggest that the overall median ranking scores between the UK medical schools differed (P=0.23; Kruskal-Wallis test). However, there was a marked difference in frequency of top 20 candidates from each medical school, which could not be explained by the size of the medical school alone. CONCLUSION: It is difficult to conclude from these findings the importance that the Duke-Elder examination has in the selection of trainees into ophthalmology. The role of cognitive factors in selection into postgraduate medical/surgical training is discussed, along with the potential academic criteria, which may influence interview scores.

Sweet-talk: role of host glycosylation in bacterial pathogenesis of the gastrointestinal tract.

Glycosylation is a key modification of proteins and lipids and is involved in most intermolecular and intercellular interactions. The gastrointestinal mucus gel is continuous and can be divided into two layers: a secreted loosely associated layer and a layer firmly attached to the mucosa. In addition, the membrane-bound glycosylated proteins and lipids create a glycocalyx, which remains adherent on each cell and is dynamic and responsive to the physiological state and environment of the cell. The secreted glycans form a mucus gel layer that serves as a physicochemical sensor and barrier network and is primarily composed of mucins and associated peptides. These glycans protect gut epithelial cells from chemical, biological and physical insults and are continuously renewed. Pathogens colonise and invade the host epithelial cells using protein-protein and glycan-lectin interactions. During the process of colonisation and infection, the glycosylation state of both host and pathogen change in response to the presence of the other. This complex modulation of glycan expression critically determines pathogenesis and the host response in terms of structural changes and immune response. In addition, by influencing host immunity and gut glycosylation, the microbiota can further effect protection against pathogens. In this review, the roles of host glycosylation in interactions with two prevalent bacterial pathogens, Campylobater jejuni and Helicobacter pylori, are discussed to illustrate important concepts in pathogenesis.

Multidrug-resistant nosocomial Citrobacter in a hospital in Kathmandu.

Citrobacter sp. is a commensal flora and an infrequent nosocomial pathogen to cause nuisance in hospital settings. Recently, the isolation of this pathogen in health care setting is rising and the multidrug resistant strains are emerging. This pathogen cause wide array of infections and the mortality rate is unexpectedly high of 30.0-60.0%. Extended spectrum cephalosporins have been used to treat this pathogen and due to the emergence of resistant strains to these drugs newer treatment protocols have to be devised. Epidemiology and antibiotic susceptibility pattern of clinical isolates of Citrobacter sp. isolated in a hospital were investigated. Specimens were collected from patients and implicated pathogens were isolated. Disk diffusion test was performed on these isolates and resistant patterns were. Antibiogram typing was used to resolve the clones of the isolated bacteria. The results showed that Citrobacter sp. was highly prevalent and commonly isolated from the sputum sample of patients diagnosed as Chronic Obstructive Pulmonary Disease (COPD). The antibiogram pattern suggested the circulation of three clones of Citrobacter sp. They were multidrug resistant and were sensitive to only cefoperazone and sulbactam suggesting the production of â-lactamse inhibitors sensitive molecular class A and D extended spectrum â-lactamases (ESBL). In conclusion, although, ESBL producers are always treated with carbapenems, we recommend to use combination therapy of â-lactam and â-lactamse inhibitors to treat this multidrug resistant Citrobacter sp. and carbapenems should be kept as a reserve drug and we should aim to prevent the spread of this resistant pathogen between different patients to decrease the morbidity and mortality associated with this pathogen.

Biocombinatorial selection of carbohydrate binding agents of therapeutic significance.

Components of the cell-surface and extracellular matrix (including cell walls of some microorganisms) present ideal but often challenging targets for disease detection and targeting. In the past few years, selection of peptide and oligonucleotide biomimics against secreted and extracellular targets has resulted in the isolation of small high affinity binding agents against target molecules. However, little attention has been paid to the interaction of these biomimics with glycans (specifically, cell surface glycans) despite their ubiquitous and abundant presence on every cell (glycocalyx). Although glycans are now accepted as critically important class of molecules as biomarkers, imaging, therapeutic and drug targets, little advance has been made in exploiting these molecules for clinical applications. The diversity and complexity of glycan structures along with time-consuming analytical and synthetic methods are among the most significant challenges faced by glycoscientists. Innovative technologies are urgently needed to overcome these challenges and to develop glycan-based clinical applications. This review article will provide an overview of existing biocombinatorial methods that focus on the selection and use of glycan-binding random oligonucleotide ligands (aptamers) as conceptually novel therapeutics for a variety of biomedical applications including anti-microbial and anti-cancer agents. Given the range and importance of protein-carbohydrate interactions in biological processes, rapid discovery of agents that mediate these interactions may provide fruitful venues towards novel therapeutics.

Space flight alters bacterial gene expression and virulence and reveals a role for global regulator Hfq.

A comprehensive analysis of both the molecular genetic and phenotypic responses of any organism to the space flight environment has never been accomplished because of significant technological and logistical hurdles. Moreover, the effects of space flight on microbial pathogenicity and associated infectious disease risks have not been studied. The bacterial pathogen Salmonella typhimurium was grown aboard Space Shuttle mission STS-115 and compared with identical ground control cultures. Global microarray and proteomic analyses revealed that 167 transcripts and 73 proteins changed expression with the conserved RNA-binding protein Hfq identified as a likely global regulator involved in the response to this environment. Hfq involvement was confirmed with a ground-based microgravity culture model. Space flight samples exhibited enhanced virulence in a murine infection model and extracellular matrix accumulation consistent with a biofilm. Strategies to target Hfq and related regulators could potentially decrease infectious disease risks during space flight missions and provide novel therapeutic options on Earth.

Marfan's syndrome with aortic valve endocarditis.

Marfans syndrome is an Autosomal dominant disorder of the connective tissues resulting in abnormalities of the musculoskeletal system, cardiovascular system and eyes. It has a prevalence of 1 in 100,000 population1 and occurs in all ethnic groups. It may be familial or due to new mutation (30%), in the fibrillin gene on arm of chromosome 15. It is estimated that one person in every 3000-5000 has Marfans syndrome may have cardiovascular abnormalities and may be complicated by infective endocartditis. About 90% of Marfan patients will develop cardiac complications2. The patient under discussion has musculoskeletal (Tall stature, reduced upper-lower segment ratio, arm-span to height ratio > 1.05, high arched palate) and Cardiovascular features (Severe aortic regurgitation complicated with infective endocarditis).

A study of the first 350 cases referred for EEG in Kathmandu Medical College Teaching Hospital.

UNLABELLED: Electroencephalography or EEG is a neurological test that uses an electronic monitoring device to measure and record electrical activity in the brain. Epilepsy is defined as a recurrent tendency to unprovoked seizure. About 0.5% to 2% of the population has epilepsy. MATERIAL AND METHODS: This is a retrospective study of the first 350 odd cases referred for EEG at Kathmandu Medical College Teaching Hospital, Kathmandu during period November 2002- September 2002 (Mangsir 2058- Bhadra 2059). RESULTS: Most patients fall in the age group of 11-20 years and more than 50% referred cases fall in the prime of life, i.e., 11-30) years. 16% of patients with the history of seizure did not use any AED and 32% who though had no history of seizures were actually using AED. It is seen that out of 149 reported normal, 57 (38%) had history of seizure; but out of 208 reported abnormal in EEG only 92 (42%) had history of seizure. DISCUSSION: EEG is not a very sensitive or a specific tool; however, in our study it was seen to be more sensitive than specific. It is till very useful in classifying seizure types, in locating epileptic focus prior to surgery or in some confusing situation when movement disorder may simulate seizure disorder or vice versa. CONCLUSION: Although EEG is not a very sensitive or specific investigation, it is more sensitive than specific.

Production of a sialylated N-linked glycoprotein in insect cells.

Under High Aspect Ratio Vessel (HARV) bioreactor culture conditions designed to simulate the microgravity of orbital space flight, insect tissue culture cells infected with a baculovirus expression vector produced a human glycoprotein with tri- and tetra-antennary complex N-linked oligosaccharides containing terminal sialic acid residues. The oligosaccharide structures were similar to those produced in human placental cells. Insect cells cultured in T-flasks only performed incomplete oligosaccharide processing. The mechanism of HARV-mediated changes in the eukaryotic N-linked glycosylation pathway was investigated and could be mimicked under T-flask growth conditions with the addition of N-acetylmannosamine to the culture medium. The significance of these investigations is discussed with respect to the production of recombinant therapeutic glycoproteins, insect physiology, and orbital space flight.

Successful establishment of long-term bone marrow cultures in 103 patients with myelodysplastic syndromes.

We used bone marrow biopsies instead of mononuclear cells to maintain long-term cultures from 103 patients belonging to all five sub-categories of myelodysplastic syndromes (MDS), as well as 12 normal controls. By week 4, 30-50% confluency was reached and could be maintained for up to 12 weeks with 100% confluency. The four prominent cells were fibroblasts, macrophages, endothelial cells and adipocytes. Immunohistochemical and electron microscopic studies provided lineage confirmation. Normal hematopoiesis was well supported by MDS stroma. Neither the FAB nor cytogenetics was co-related with the potency of growth. MDS stroma appears to be both morphologically and functionally normal.

Characterization of glycosylated variants of beta-lactoglobulin expressed in Pichia pastoris.

Glycosylated variants of beta-lactoglobulin (BLG) were produced in the methylotrophic yeast Pichia pastoris to mimic the glycosylation pattern of glycodelin, a homologue of BLG found in humans. Glycodelin has three sites for glycosylation, corresponding to amino acids 63-65 (S1), 85-87 (S2) and 28-30 (S3) of BLG. These three sites were engineered into BLG to produce the variants S2, S12 and S123, which carried one, two and three glycosylation sites, respectively. The oligosaccharides on these BLG variants ranged from (mannose)(9)(N-acetylglucosamine)(2) (Man(9)GN(2)) to Man(15)GN(2) and were of the alpha-linked high mannose type. The variant S123 exhibited highest levels of glycosylation, with the range of glycans being Man(9-14)GN(2). Digestion of S123 with alpha-1,2 linkage specific mannosidase resulted in a single product corresponding to Man(6)GN(2). These results indicated a glycosylation pattern consisting of a Man(5)GN(2) structure extended by 4-9 mannose residues attached mainly by alpha-1,2 linkages. The results also indicated extension of the Man(5)GN(2) structure by a single alpha-1,6-linked mannose. The N-linked glycosylation pathway in P.pastoris is significantly different from that in Saccharomyces cerevisiae, with the addition of shorter outer chains to the core and no alpha-1,3 outer extensions.