Consenso colombiano de expertos sobre recomendaciones basadas en evidencias para el diagnóstico, el tratamiento y el seguimiento de la enfermedad de Fabry con compromiso renal

Contexto: la enfermedad de Fabry se comporta como una enfermedad crónica con compromiso multisistémico y alto costo en salud. Objetivo: generar recomendaciones basadas en la evidencia para el diagnóstico, el tratamiento y el seguimiento de la enfermedad de Fabry con compromiso renal mediante un consenso de expertos. Metodología: a partir de la búsqueda de evidencia en Pubmed, Embase y Google Scholar entre 2010 y agosto 2020, se formulan recomendaciones sobre la definición, el diagnóstico y el tratamiento de la enfermedad de Fabry en población adulta, las cuales se consultan a un panel de expertos a través de la metodología de consenso Delphi modificado. La calidad de los documentos se evaluó por equipo metodológico aplicando herramientas en función del tipo de documento incluido. Resultados: se formularon 53 recomendaciones sobre la definición, el diagnóstico y el tratamiento. Un panel de cinco expertos clínicos nacionales e internacionales externos al grupo desarrollador participaron en la consulta preconsenso y 50 recomendaciones fueron acordadas para su inclusión, para tres de ellas se requirió una sesión formal de consenso que se dio en una ronda, incorporando tres nuevas recomendaciones. Conclusiones: las recomendaciones basadas en evidencia y experticia clínica permitirán orientar de manera estandarizada a nivel nacional y regional, el diagnóstico y el tratamiento de pacientes con sospecha o enfermedad de Fabry con compromiso renal.

Background: Fabry disease behaves like a chronic condition, with multisystem involvement and high health care costs. Objective: To generate evidence-based recommendations for the diagnosis, treatment and follow-up of the Anderson-Fabry disease with renal commitment, through an expert consensus. Methodology: Based on the search of evidence in PubMed, Embase and Google Scholar between 2010 and August, 2020, recommendations on the definition, diagnosis and treatment of Fabry Disease in adult population were formulated after consulting with an expert panel through the modified Delphi consensus methodology. The quality of the documents was assessed by methodological team applying tools according to the type of document included. Results: 53 recommendations for the definition, diagnosis and treatment were formulated. A panel of five national and international clinical experts external to the developer group participated in the pre-consensus consultation and 50 recommendations were agreed upon for their inclusion. For 3 recommendations, a formal consensus session which took place in one round was required, and 3 new recommendations were incorporated. Conclusions: The recommendations based on evidence and clinical expertise will allow us to guide the diagnosis and treatment of patients with Fabry disease with renal involvement or suspicion thereof in a standardized manner at national and regional levels.

Incidencia de la enfermedad de Fabry en pacientes con enfermedad renal crónica atendidos en tres centros de salud del departamento de Atlántico, Colombia. 2017-2018

Introducción: la enfermedad de Fabry es un defecto lisosomal caracterizado por una alteración de la enzima alfa-galactosidasa y que produce el acúmulo de glucoesfingolípidos en diferentes tejidos. Este defecto enzimático está ligado al cromosoma X y por ende es más frecuente en hombres. Sus manifestaciones clínicas varían de acuerdo al grupo etario afectado e incluyen lesiones en piel, anhidrosis, opacidades corneales, crisis de dolor, daño renal, entre otros. Objetivo: calcular la incidencia de enfermedad de Fabry en pacientes con diagnóstico de enfermedad renal crónica (ERC). Materiales y métodos: estudio ambispectivo realizado en los pacientes con diagnóstico de ERC que asistían a controles médicos en tres centros de prevención renal ubicados en el departamento del Atlántico, Colombia, y que además cumplían los criterios de inclusión y exclusión. Se revisaron las historias clínicas y se tomaron muestras para confirmar la presencia de enfermedad de Fray. Resultados: se identificaron 471 pacientes con ERC y se estableció una incidencia global de 21,23 casos por cada 1.000 habitantes para baja actividad de la alfa-galactosidasa. Sin embargo, solo en el 20% se confirmó la presencia de enfermedad de Fray mediante pruebas genéticas. Conclusiones: la incidencia de la enfermedad de Fabry en la población estudiada es mayor a la reportada en otras cohortes y además fue más frecuente en el sexo femenino.

Introduction: Fabry's disease consists of a lysosomal defect linked to the X chromosome that produces the accumulation of glycosphingolipids in different tissues. The clinical manifestations depend on the age of presentation, and includes skin lesions, acroparesthesia, pain crisis, anhidrosis, corneal opacities and hearing loss, among others. Objectives: Calculate the incidence of Fabry disease in patients diagnosed with chronic kidney disease Methodology: An ambispective study was designed, including all patients diagnosed with chronic kidney disease under medical control in three renal prevention centers located in the department of Atlántico, and which also met the inclusion and exclusion criteria. Subsequently, the review of the medical records and the sampling were carried out. Results: A total of 471 patients with chronic kidney disease were identified, with an overall incidence of 21.23 cases per 1000 people. However, only 20% were confirmed by genetic tests. Conclusions: The incidence of Fabry disease in the population studied is greater than that reported in other cohorts. In addition, it is more frequent in the female sex.

Role of glycosylation in TGF-β signaling and epithelial-to-mesenchymal transition in cancer

Glycosylation is a common posttranslational modification on membrane-associated and secreted proteins that is of pivotal importance for regulating cell functions. Aberrant glycosylation can lead to uncontrolled cell proliferation, cell-matrix interactions, migration and differentiation, and has been shown to be involved in cancer and other diseases. The epithelial-to-mesenchymal transition is a key step in the metastatic process by which cancer cells gain the ability to invade tissues and extravasate into the bloodstream. This cellular transformation process, which is associated by morphological change, loss of epithelial traits and gain of mesenchymal markers, is triggered by the secreted cytokine transforming growth factor-β (TGF-β). TGF-β bioactivity is carefully regulated, and its effects on cells are mediated by its receptors on the cell surface. In this review, we first provide a brief overview of major types of glycans, namely, N-glycans, O-glycans, glycosphingolipids and glycosaminoglycans that are involved in cancer progression. Thereafter, we summarize studies on how the glycosylation of TGF-β signaling components regulates TGF-β secretion, bioavailability and TGF-β receptor function. Then, we review glycosylation changes associated with TGF-β-induced epithelial-to-mesenchymal transition in cancer. Identifying and understanding the mechanisms by which glycosylation affects TGF-β signaling and downstream biological responses will facilitate the identification of glycans as biomarkers and enable novel therapeutic approaches.

Review and Proposal of Alternative Technologies for Comprehensive and Reliable Newborn Screening Using Paper Borne Urine Samples for Lysosomal Storage Disorders: Glycosphingolipid Disorders

Abstract Few current methods are efficient to detect a high number of lysosomal storage disorders (LSDs) in newborn screening. Therefore, we propose a stepwise procedure that starts with the use of paper borne urine samples (Berry-Woolf specimen) for the inexpensive detection of elevated lysosomal content and the identification of which of the three majors biochemical groups -mucopolysaccharides, oligosaccharides, and glycosphingolipids- is detected. Urine samples are preferable to blood samples because of their higher concentrations of the relevant analytes. Subsequent steps would precisely determine which enzyme deficiency is involved. As a summary, following our previous papers on the detection of elevated oligosaccharides and mucopolysaccharides, here we describe how elevated urinary glycosphingolipids (GSLs) could be fluorometrically detected using the reagent 5-hydroxy-1-tetralone (HOT) and subsequently identified with precision by continuous thin layer chromatography or other techniques. We also outline the steps required for the validation of this procedure for its introduction in newborn screening programs.

GLA Gene Mutation in Hypertrophic Cardiomyopathy with a New Variant Description: Is it Fabry's Disease? / Mutação do Gene GLA na Cardiomiopatia Hipertrófica com Descrição de Nova Variante: É Doença de Fabry?

Abstract Background: Fabry disease (FD) is an X-linked lysosomal storage disorder caused by mutations in the alpha galactosidase A gene (GLA) that lead to the enzymatic deficiency of alpha galactosidase (α-Gal A), resulting in the accumulation of globotriaosylceramide (Gb3) and globotriaosylsphingosine (lyso-Gb3), causing multiple organ dysfunctions. Objective: To perform GLA gene screening in a group of patients with echocardiographic diagnosis of hypertrophic cardiomyopathy (HCM). Methods: a cross-sectional study was conducted with HCM patients from a university hospital. Patients with coronary artery disease and valvulopathies were excluded. Mutation analysis of the GLA gene was performed. In male subjects, the analysis was performed after evidence of low α-Gal A activity. Results: 60 patients with echocardiographic diagnosis of HCM were included. Age ranged from 12 to 85 years and 60% were women. Mean myocardial fibrosis percentage on MRI was 10.7 ± 13.1% and mean ventricular thickness was18.7 ± 6.7 mm. Four patients had the following GLA gene mutations: c.967C>A (p.Pro323Thr), not yet described in the literature; c.937G>T (p.Asp313Tyr); and c.352C>T (p.Arg118Cys). All patients had normal levels of lyso-Gb3 and non-ischemic myocardial fibrosis on magnetic resonance imaging; one patient had proteinuria and one patient had ventricular tachycardia. Conclusion: in this study, the frequency of mutation in the GLA gene in patients with HCM was 6.7%. A novel mutation in exon 6 of the GLA gene, c.967C>A (p.Pro323Thr), was identified. Patients with HCM may have GLA mutations and FD should be ruled out. Plasma (lyso-Gb3) levels do not seem to be sufficient to attain a diagnosis and organ biopsy should be considered.

Resumo Fundamento: A doença de Fabry (DF) é uma doença de armazenamento lisossômico ligada ao cromossomo X, devido a mutações no gene da alfa galactosidase A (GLA), levando a deficiência enzimática de alfa-galactosidase (α-Gal A) e acúmulo de globotriaosilceramida (Gb3) e globotriaosilsulfingosina (liso-Gb3), causando disfunção de múltiplos órgãos. Objetivo: realizar a triagem do gene GLA em um grupo de pacientes com diagnóstico ecocardiográfico de cardiomiopatia hipertrófica (CMH). Métodos: estudo transversal realizado com pacientes com CMH em um hospital universitário. Pacientes com doença arterial coronariana e valvopatias foram excluídos. Foi realizada análise de mutação do gene GLA. Em indivíduos do sexo masculino, a análise foi realizada após evidência de baixa atividade de α-Gal A. Resultados: Foram incluídos 60 pacientes com diagnostico ecocardiográfico de CMH. A idade variou de 12 a 85 anos e 60% eram mulheres. O percentual médio de fibrose miocárdica na RM foi 10,7 ± 13,1% e a espessura ventricular média foi 18,7 ± 6,7 mm. Quatro pacientes tinham as seguintes mutações do GLA: c.967C>A (p.Pro323Thr), ainda não descrita na literatura; c.937G>T (p.Asp313Tyr); e c.352C>T (p.Arg118Cys). Todos os pacientes apresentavam níveis normais de liso-Gb3 e fibrose miocárdica não isquêmica na ressonância magnética; um paciente apresentou proteinúria; um paciente apresentou taquicardia ventricular. Conclusão: Neste estudo, a frequência de mutação no gene GLA em pacientes com CMH foi 6,7%. Uma nova mutação no exon 6 do gene GLA, c.967C>A (p.Pro323Thr), foi identificada. Pacientes com CMH podem ter mutações do GLA e a DF deve ser excluída. Os níveis plasmáticos de (liso-Gb3) não parecem ser suficientes para fazer um diagnóstico e biópsia de órgãos deve ser considerada.

Therapeutic effect of intravenous drip of MTG combined comprehensive rehabilitation on patients with stroke hemiplegia and its impact on neurologic impairment score／ / 心血管康复医学杂志

To discuss therapeutic effect of intravenous drip of monosialic tetrahexose ganglioside (MTG) combined comprehensive rehabilitation on patients with stroke hemiplegia and its impact on neurologic impairment score (NIS).Methods : A total of 100 patients with stroke hemiplegia treated in our hospital were randomly and e‐qually divided into routine treatment group (received routine comprehensive training treatment including work ther‐apy ,exercise therapy etc .) and combined treatment group (received intravenous drip of MTG based on routine treatment group) ,both groups were continuously treated for four weeks .Limb and neurologic function ,and im‐provement of quality of life (QOL) were compared between two groups .Results : Oneight weeks after treatment , total effective rate of limb function in combined treatment group was significantly higher than that of routine treat‐ment group (94. 0% vs.76. 0%, P＝0.030) ;after one ,four and eight‐week treatment ,along with time went on , there was significant rise in Glasgow coma scale (GCS) score in two groups ,and those of combined treatment group were significantly higher than those of routine treatment group [one week :(8. 56 ± 2. 47) scores vs .(7.24 ± 2.01) scores ,four weeks :(11.26 ± 2.81) scores vs .(9.23 ± 2. 01) scores ,eight weeks :(13.28 ± 3.43) scores vs .(11. 43 ± 2.65) scores] , P＜0. 01 all ;compared with before treatment ,there were significant rise in each dimension score of QOL in two groups after eight‐week treatment ,and each dimension score of combined treatment group was sig‐nificantly higher than that of routine treatment group , P＝0. 001 all.Conclusion : Intravenous drip of MTG com‐bined comprehensive rehabilitation possesses significant therapeutic effect on patients with stroke hemiplegia .It can significantly improve limb and neurologic function state ,improve quality of life in these patients .

Glycosphingolipid Modification: Structural Diversity, Functional and Mechanistic Integration of Diabetes

Glycosphingolipids (GSLs) are present in all mammalian cell plasma membranes and intracellular membrane structures. They are especially concentrated in plasma membrane lipid domains that are specialized for cell signaling. Plasma membranes have typical structures called rafts and caveola domain structures, with large amounts of sphingolipids, cholesterol, and sphingomyelin. GSLs are usually observed in many organs ubiquitously. However, GSLs, including over 400 derivatives, participate in diverse cellular functions. Several studies indicate that GSLs might have an effect on signal transduction related to insulin receptors and epidermal growth factor receptors. GSLs may modulate immune responses by transmitting signals from the exterior to the interior of the cell. Guillain-Barre syndrome is one of the autoimmune disorders characterized by symmetrical weakness in the muscles of the legs. The targets of the immune response are thought to be gangliosides, which are one group of GSLs. Other GSLs may serve as second messengers in several signaling pathways that are important to cell survival or programmed cell death. In the search for clear evidence that GSLs may play critical roles in various biological functions, many researchers have made genetically engineered mice. Before the era of gene manipulation, spontaneous animal models or chemical-induced disease models were used.

Current relevance of fungal and trypanosomatid glycolipids and sphingolipids: studies defining structures conspicuously absent in mammals

Recently, glycosphingolipids have been attracting attention due to their role on biological systems as second messengers or modulators of signal transduction, affecting several events, which range from apoptosis to regulation of the cell cycle. In pathogenic fungi, glycolipids are expressed in two classes: neutral monohexosylceramides (glucosyl-or galactosylceramide) and acidic glycosylinositol phosphorylceramides (the latter class carries longer glycan chains). It is worth to mention that monohexosylceramides exhibit significant structural differences in their lipid moieties compared to their mammalian counterparts, whereas the glycosylinositol phosphorylceramides exhibit remarkable structural differences in their carbohydrate moieties in comparison to mammal glycosphingolipids counterpart. We observed that glycosylinositol phosphorylceramides are capable of promoting immune response in infected humans. In addition, inhibiting fungal glycosphingolipid biosynthetic pathways leads to an inhibition of colony formation, spore germination, cell cycle, dimorphism and hyphal growth. Other pathogens, such as trypanosomatids, also present unique glycolipids, which may have an important role for the parasite development and/or disease establishment. Regarding host-pathogen interaction, cell membrane rafts, which are enriched in sphingolipids and sterols, participate in parasite/fungal infection. In this review, it is discussed the different biological roles of (glyco) (sphingo) lipids of pathogenic/opportunistic fungi and trypanosomatids.

Recentemente, glicoesfingolipídeos têm atraído atenção devido ao seu papel na biologia celular como segundo-mensageiro ou moduladores da transdução de sinal, afetando vários eventos, desde apoptose até a regulação do ciclo celular. Em fungos patogênicos, existem duas classes de glicolipídeos: monohexosil ceramidas neutras (glucosil-ou galactosilceramida) e glicosilinositol fosforilceramidas (os quais apresentam cadeias de carboidratos mais longas). É importante enfatizar que as monohexosil ceramidas exibem diferenças estruturais nas suas porções lipídicas quando comparadas às de mamíferos, enquanto que glicosilinositol fosforilceramidas exibem diferenças estruturais marcantes em suas porções carboidratos em comparação aos glicoesfingolipídeos de mamíferos. Observamos também que glicosilinositol fosforilceramidas são capazes de promover resposta imune em indíviduos infectados. Além do mais, inibição das vias biossintéticas de glicoesfingolipídeos de fungos acarreta a inibição da formação de colônias, germinação de esporos, ciclo celular, dimorfismo e crescimento de hifas. Outros patógenos, como os tripanosomatídeos, também apresentam glicolipídeos únicos, os quais apresentam um papel importante para o desenvolvimento do parasita e/ou para o estabelecimento da doença. Em relação à interação hospedeiro-patógeno, os "membrane rafts", estruturas da membrana plasmática enriquecidas em esfingolipídeos e esteróis, têm participação fundamental na infecção do parasita/fungo. Nesta revisão, discutimos os diferentes papéis biológicos dos (glico) (esfingo) lipídeos de fungos patogênicos/oportunistas e de tripanosomatídeos.

The roles of glycosphingolipids in the proliferation and neural differentiation of mouse embryonic stem cells

Glycosphingolipids including gangliosides play important regulatory roles in cell proliferation and differentiation. UDP-glucose:ceramide glucosyltransferase (Ugcg) catalyze the initial step in glycosphingolipids biosynthesis pathway. In this study, Ugcg expression was reduced to approximately 80% by short hairpin RNAs (shRNAs) to evaluate the roles of glycosphingolipids in proliferation and neural differentiation of mouse embryonic stem cells (mESCs). HPTLC/immunofluorescence analyses of shRNA-transfected mESCs revealed that treatment with Ugcg-shRNA decreased expression of major gangliosides, GM3 and GD3. Furthermore, MTT and Western blot/immunofluorescence analyses demonstrated that inhibition of the Ugcg expression in mESCs resulted in decrease of cell proliferation (P < 0.05) and decrease of activation of the ERK1/2 (P < 0.05), respectively. To further investigate the role of glycosphingolipids in neural differentiation, the embryoid bodies formed from Ugcg-shRNA transfected mESCs were differentiated into neural cells by treatment with retinoic acid. We found that inhibition of Ugcg expression did not affect embryoid body (EB) differentiation, as judged by morphological comparison and expression of early neural precursor cell marker, nestin, in differentiated EBs. However, RT-PCR/immunofluorescence analyses showed that expression of microtubule- associated protein 2 (MAP-2) for neurons and glial fibrillary acidic protein (GFAP) for glial cells was decreased in neural cells differentiated from the shRNA-transfected mESCs. These results suggest that glycosphingolipids are involved in the proliferation of mESCs through ERK1/2 activation, and that glycosphingolipids play roles in differentiation of neural precursor cells derived from mESCs.

Glycoconjugates and polysaccharides of fungal cell wall and activation of immune system / Glicoconjugados e polissacarídios de parede celular de fungos e ativação do sistema imune

The fate of organochlorine 14C-dicofol in activated sludge process was investigated. Results showed that the major part of radioactivity remained adsorbed on biological sludge. Consequently, its final disposal deserves special attention. The small amounts of dicofol, biotransformed or not, which remained in the treated effluent could contaminate receiving bodies.

Glicoproteínas, glicoesfingolipídios e polissacarídios, expostos nas camadas mais externas da parede celular dos fungos, estão envolvidos em diferentes tipos de interações com o ambiente extracelular. Essas moléculas são componentes essenciais desses organismos, contribuindo para a estrutura, integridade, crescimento celular, diferenciação e sinalização. Alguns são compostos imunologicamente ativos com potencial para regular a patogênese e a resposta imune do hospedeiro, Algumas dessas estruturas podem ser especificamente reconhecidas por anticorpos presentes no soro de pacientes, sugerindo uma possível utilização como ferramenta no diagnóstico das infecções fúngicas.

Fabry's disease: a case report and review of literatures reported in Korea

Fabry's disease is a rare, X-linked disorder of the glycosphingolipid metabolism, in which a partial or total deficiency of a lysosomal alpha(alpha)-galactosidase results in the progressive accumulation of neutral glycosphingolipids with terminal alpha galactose moieties (i.e., cerebroside di- and trihexoside) in most body fluids and tissues. Accumulation of neutral glycosphingolipids occurs within the lysosomes of endothelial, perithelial, and smooth muscle cells of the myocardial and renal systems; to a lesser extent in reticuloendothelial and connective cells of the cornea; and in ganglion and perineural cells of the autonomic nervous system. In Korea, 7 cases of Fabry's disease have been reported. A 29-year-old man with fever and headache had typical skin findings and a family history of Fabry's disease, and it was confirmed through renal biopsy and enzyme assay for alpha-galactosidase. We report a case of Fabry's disease with a review of the literatures reported in Korea.

Biosynthesis of eukaryotic cell surface glycosphingolipids using solubilized glycosyltransferases.

Two fucsyltransferases (FucT-2 and FucT-3) have been solubilized from Golgi-rich membrane fraction of bovine spleen, using a cationic detergent. FucT-3 was distinguished from FucT-2 by comparing their kinetic parameters and heat stability. FucT-2 and FucT-3 lost activity (85 %) and (5 %), respectively, when heated at 55°C for 10 sec. Two galactosyltransferases (GalT-3 and GalT-4) and two sialyltransferases (SAT-2 and SAT-3) have also been solubilized from embryonic chicken brain membranes using nonionic detergents. Affinity chromatography and microisoelectric focusing were used to separate these enzymes into functionally pure fractions. Anomeric and positional linkages in some of the products (LM1 and LD1c) have also been established. The terminal NeuAc(α2–8) linkage in GD3 and LD1c was established by identification of the partially methylated penultimate [Ac-14C]sialic acid.

A NEW METHOD OF VISUALIZING NEUTRAL GLYCOSPHINGOLIPIDS / 解放军医学杂志

To investigate a simple method visualizing neutral glycosphingolipid (N-GSLs) with iodine vapor, N-GSLs were isolated from the normal brain tissue of human by a modificated method of Hakomori, and then analyzed by HPTLC and visualized respectively by spraying aniline-diphenylamine-H 3PO 4 reagent and immersing into a jar full of iodine vapor. The plate should be kept 110℃ for 5 min until the coloration emerged by using the first method,and the N-GSLs were slowly made visible with a blue coloration. The N-GSLs were immediately made visible with a red-brown coloration by using the second method, and the color visible on the plates could emerge again after vaporized. Moreover, the second method was more sensitive than the first. The location of individual N-GSLs was the same by using the two methods. It suggested that a new method for visualizing N-GSLs has been established, it is simple, economic, rapid, innocuity and more sensitive, and the results are reproducible.

CMH IS THE MULTIDRUG RESISTANCE(MDR) ASSOCIATED NEUTRAL GLYCOSPHINGOLIPID IN KBv_(200)CELL LINE / 解放军医学杂志

In order to study the relationship between the expression of neutral glycosphingolipids(N-GSLs) and MDR, N-GSLs in MDR cell line KBv 200 and its parental cell line KB were studied. The N-GSLs were extracted and purified from the KB and KBv 200 cells according to the modified method of Hakomori, then analysed by High Performance Thin Layer Chromatography (HPTLC). The effects of PPMP on the expression of N-GSLs in KBv 200 cell line were also studied by the above method, its reversion on the KBv 200 cells to Vincristine (VCR) was examined by MTT method. The results showed that the levels of CMH and CDH in KBv 200 cells were higher than in KB cells, CMH was much more markedly increased, PPMP could reverse MDR by inhibiting CMH synthesis. It suggested that CMH is MDR associated N-GSLs in KBv 200 cell line, and inhibition of CMH expression in tumor cells maybe a new way to reverse MDR.

INFLUENCE OF TUMOR MDR ASSOCIATED CMH ON THE EXPRESSION OF B7 IN DC / 解放军医学杂志

To investigate the role of tumor multidrug resistance (MDR) associated with CMH in the immunological evasion of MDR tumor cells, CMH was separated from neutral glycolipids of KBv 200by column chromatography, and the expression of B7 in DC was detected by flow cytometry. The results showed that the expression of B7 in DC was inhibited by CMH, it suggested the tumor MDR associated with glycolipids may inhibit human immune response by its effects on DC.

REGULATION OF EXPRESSION OF N-GSLS AND THE REVERSION OF MDR IN OVARIAN CANCER CELLS (COC1/DDP) / 解放军医学杂志

In order to evaluate the relationship between MDR of ovarian cancer and expression of neutral glycosphingolipids(N-GSLs) in ovarian cancer cell lines, the effects of TAM and VRP on the growth of COC1/DDP were assayed by MTT method. N-GSLs of the cells were isolated and purified with the modified Hakamoris method and analysed by HPTLC. The results showed that the expression of N-GSLs was different between parent cell lines and resistant cell sublines, the level of CMH was higher in COC1/DDP than in COC1. TAM and VRP could render multidrug-resistant cells sensitive to chemotherapy, while the level of CMH concomitantly was sharply decreased. It suggested that the expression of N-GSLs is associated with MDR of ovarian cancer, and CMH may be a kind of MDR related glycolipids in ovarian cancer.

PRELIMINARY STUDY OF THE MOLECULAR STRUCTURE OF GLOB-N:A GLIOMA INHIBITOR / 解放军医学杂志

In order to study the molecular structure of Glob-N, a glioma inhibitor, Glob-N was separated from the membrane of human type O red blood cells, cow brain and normal human brain tissue respectively. Their structures were identified by using time-of-flight mass spectrometry, 1H-NMR and 13C-NMR. The results showed that the structure of Glob-N from the membrane of human type O red blood cells was proposed as globotetraosylceramide, while Glob-N from cow brain and normal human brain was globotriaoceramide, and the number of carbon atoms in the ceramide of the Glob-N was 27, 33 and 26 respectively. It suggested that Glob-N is a kind of neutral glycosphingolipids, and its molecular structure is different in different tissues.

MODULATION OF MDR1 mRNA EXPRESSION AND REVERSION OF MULTI-DRUG RESISTANCE BY PPMP IN KBv_(200) CELL LINE / 解放军医学杂志

The current study was designed to investigate the effects of PPMP (DL-threo-1-phenyl-2- palmitoylamino-3-morpholino-1-propanol), a kind of glycolipids synthase inhibitor, on the modulation of mdr1 mRNA expression and the reversing effect of multi-drug resistance by PPMP in human malignancy KBv 200cell line. In vitro KBv 200 cells were treated with PPMP in different concentration, the alterations of mRNA expression of drug-resistant gene mdr1 in KB (sensitive cell line) and KBv 200 (before and after the treatment of PPMP) cells were analyzed by RT-PCR. Intracellular rhodamine(Rh123) concentration was measured by flow cytometry. PPMP was found to inhibit mdr1 gene expression of KBv 200 at the mRNA level, and complete inhibition appeared at 25?mol/L PPMP treatment for 48h. PPMP could increase intracellular Rh123 accumulation in resistant cell lines. This modulation of gene expression and Rh123 accumulation was directly correlated with the concentration of PPMP. It suggested that PPMP, a chemical inhibitor of glycolipids synthase, could modulate mdr1 expression at the mRNA level in a content dependent manner, PPMP possesses MDR-reversing activity.