Preparation of novel gypenosides long-circulating liposomes consisted by sphingomyelin and beta-sitosterol modified by PEG / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the feasibility of preparing novel gypenosides long-circulating liposomes with PEG grafted on beta-sitosterol (PEG-Sito).</p><p><b>METHOD</b>Succinicanhydride was adopted to connect beta-sitosterol and PEG 2000. Sphingomyelin and PEG-Sito was used as material to prepare gypenosides long-circulating liposomes by using ethanol injection method. Encapsulation efficiency was determined by using protamine precipitation method. H-NMR was used to verify the synthesis of PEG-Sito, the novel gypenosides long-circulating liposomes were characterized by particle size, zeta potential and atomic force microscope.</p><p><b>RESULT</b>The synthesis of PEG-Sito was verified by 1H-NMR. Encapsulation efficiency of long-circulating liposomes prepared by ethanol injection method was 74.3%, particle size was 288.1 nm, zeta potential was -20.25 mV, the morphology were round observed by AFM.</p><p><b>CONCLUSION</b>The novel gypenosides long-circulating liposomes prepared with PEG-Sito was feasible, it had a high encapsulation efficiency and good morphology.</p>

Study on preparation of gypenosides liposomes composed of sphingomyelin and cholesterol and its quality evaluation / 中国中药杂志

<p><b>OBJECTIVE</b>To discuss the feasibility of preparing gypenosides liposomes with sphingomyelin and cholesterol, and optimize the preparation process and prescription.</p><p><b>METHOD</b>Gypenosides liposomes were prepared with sphingomyelin and cholesterol. The entrapment efficiency was determined by the protamine precipitation method. The entrapment efficiency was taken as the evaluation index to screen out the optimum preparation process for the new-type gypenosides liposomes. The preparation process was optimized by the orthogonal design. The new-type gypenosides liposomes were characterized by grain size, potential and atomic force microscope (AFM).</p><p><b>RESULT</b>Ethanol injection was the optimum process to prepare gypenosides liposome with sphingomyelin and cholesterol as follow: the ratio of gypenosides to sphingomyelin was 1: 10, the ratio of sphingomyelin to cholesterol was 4: 1, the pH of PBS buffer solution was 7.0, the hydration temperature was 45 degrees C, the entrapment efficiency was 79.06%, particle size was 191.4 nm, the Zeta potential was -33.16 mV, and the morphology were round under AFM.</p><p><b>CONCLUSION</b>It was feasible to prepare gypenosides liposome with sphingomyelin and cholesterol. The gypenosides liposomes prepared by the optimum preparation process were good in morphology, particle size and reproducibility.</p>

Analysis of Acid Sphingomyelinase Activity in Dried Blood Spots Using Tandem Mass Spectrometry

BACKGROUND: Niemann Pick disease (NP) is a rare, lysosomal storage disorder due to deficiency of the intra-lysosomal enzyme acid sphingomyelinase (ASM) resulting in intracellular accumulation of sphingomyelin. We evaluated a tandem mass spectrometry (MS/MS) method to analyze ASM activity in dried blood spots (DBS) that may be suitable for laboratory diagnosis of NP including high throughput screening of at-risk populations and potentially for newborn screening. METHODS: ASM activity was measured in 3.2 mm punches from DBS. The eluate was incubated with the ASM substrate (N-Hexanoyl-D-erythro-sphingosylphosphorylcholine [C6-sphingomyelin (C29H59N2O6P)]) and an internal standard (N-butyroyl-D-erythro-sphingosine [C4-ceramide (C22H43NO3)]). ASM product and IS were analyzed using MS/MS in multiple reaction monitoring mode for transitions m/z 370.6>264.3 (ASM internal standard) and m/z 398.6>264.3 (ASM product). RESULTS: ASM activities were stable for up to 2 months at or below 4degrees C. Position of the punch in the DBS and/or hematocrit of the DBS had a limited effect on ASM activities. Both intra- and inter-assay variability were below 10%. There was no carry-over. The median ASM activity in 2,085 newborn infants was 9.5 micromol/h/L (mean 10.6) with a SD of 5.06 micromol/h/L. Six of 2,085 (0.3%) infants were found to have ASM activities below the cut-off of 2.5 micromol/h/L. ASM activities were below the cut-off level in all 10 previously diagnosed cases with NP (range: 0.16 to 2.08 micromol/h/L). CONCLUSIONS: This MS/MS method for the measurement of ASM activity in DBS is robust and suitable for laboratory diagnosis of NP.

Analysis of Acid Sphingomyelinase Activity in Dried Blood Spots Using Tandem Mass Spectrometry

BACKGROUND: Niemann Pick disease (NP) is a rare, lysosomal storage disorder due to deficiency of the intra-lysosomal enzyme acid sphingomyelinase (ASM) resulting in intracellular accumulation of sphingomyelin. We evaluated a tandem mass spectrometry (MS/MS) method to analyze ASM activity in dried blood spots (DBS) that may be suitable for laboratory diagnosis of NP including high throughput screening of at-risk populations and potentially for newborn screening. METHODS: ASM activity was measured in 3.2 mm punches from DBS. The eluate was incubated with the ASM substrate (N-Hexanoyl-D-erythro-sphingosylphosphorylcholine [C6-sphingomyelin (C29H59N2O6P)]) and an internal standard (N-butyroyl-D-erythro-sphingosine [C4-ceramide (C22H43NO3)]). ASM product and IS were analyzed using MS/MS in multiple reaction monitoring mode for transitions m/z 370.6>264.3 (ASM internal standard) and m/z 398.6>264.3 (ASM product). RESULTS: ASM activities were stable for up to 2 months at or below 4degrees C. Position of the punch in the DBS and/or hematocrit of the DBS had a limited effect on ASM activities. Both intra- and inter-assay variability were below 10%. There was no carry-over. The median ASM activity in 2,085 newborn infants was 9.5 micromol/h/L (mean 10.6) with a SD of 5.06 micromol/h/L. Six of 2,085 (0.3%) infants were found to have ASM activities below the cut-off of 2.5 micromol/h/L. ASM activities were below the cut-off level in all 10 previously diagnosed cases with NP (range: 0.16 to 2.08 micromol/h/L). CONCLUSIONS: This MS/MS method for the measurement of ASM activity in DBS is robust and suitable for laboratory diagnosis of NP.

Effect of tangshen formula on phospholipids metabolism in diabetic nephropathy patients / 药学学报

This study was to report the effect of Tangshen Formula on phospholipids metabolism in diabetic nephropathy patients. A normal phase-HPLC-TOF/MS method was used in this study for the determination of seven species of phospholipids in human plasma. Then, the concentration changes of potential phospholipids biomarkers were discussed in diabetic nephropathy phase III and phase IV patients among different groups, including before and 3, 6 months after administration of Tangshen Formula. Significant increases of PE750, PI885, PC792, PC826, PC830, PC854 and PC802 levels were observed 6 months after administration of Tangshen Formula and conventional western medicine, as well as a decrease of LPC540 level, when compared with those before medication. Concentrations of all the potential phospholipids biomarkers showed a tendency towards normal levels; however, both the improvement degree and onset time of these compounds were not same. Additionally, Tangshen Formula treatment based on conventional western medicine treatment was more efficient in adjusting the levels of these compounds when compared with western medicine treatment alone, especially for the phase IV patients. These results indicated that Tangshen Formula was capable in regulating and improving phospholipids metabolism in diabetic nephropathy patients, which may be related with the direct or indirect inhibition of protein kinase C pathway and the corresponding reduction of phospholipase A2 activity. Therefore, Tangshen Formula may be used as an effective drug for diabetic nephropathy therapy, at least as an adjunctive therapeutic drug.

Sphingomyelin synthase 2 deficiency decreases atherosclerosis and inhibits inflammation in mice / 生理学报

Plasma sphingomyelin (SM) has been shown to be an independent risk factor for coronary heart disease, and sphingomyelin synthase 2 (SMS2) contributes to de novo SM biosynthesis and plasma membrane SM levels. The aim of the present study is to evaluate the in vivo role of SMS2 deficiency in serum SM metabolism and atherosclerosis (AS) development. We used male SMS2 knockout (SMS2(-/-)) and C57BL/6J (wild-type, WT) mice as experimental and control groups, respectively. Each group was fed high-fat diet (1% cholesterol, 20% leaf fat), as well as bile salt for accelerating the atherosclerotic formation. After three months of feeding, the mice were killed to observe aortic arches and oil red-stained longitudinal sections of thoracoabdominal aortae. Fasting blood samples were taken from the tail vein before and after high-fat diet, and the serum lipid and SM levels were measured by using kits and enzymatic method respectively. Western blot was used to analyze the contents of nuclear factor-kappaB (NFkappaB) p65 subunit in peritoneal macrophages stimulated with lipopolysaccharide (LPS) after high-fat diet. The results showed that after high-fat diet, SMS2(-/-) mice presented decreased atherosclerotic lesions in aortic arch and thoracoabdominal aorta compared with WT mice. Regardless of whether high-fat diet were given or not, SMS2(-/-) mice showed a significant decrease in serum SM level (P<0.05), but no significant changes in serum lipid levels, compared with WT mice. The expressions of NFkappaB p65 were attenuated in macrophages from SMS2(-/-) mice in response to LPS stimulation compared with those of the WT mice. These results suggest that SMS2 deficiency decreases AS and inhibits inflammation in mice. Thus, SMS2 deficiency may be a potential therapeutic strategy.

Nerve growth factor, sphingomyelins, and sensitization in sensory neurons / 生理学报

Because nerve growth factor (NGF) is elevated during inflammation, plays a causal role in the initiation of hyperalgesia, and is known to activate the sphingomyelin signalling pathway, we examined whether NGF and its putative second messenger, ceramide, could modulate the excitability of capsaicin-sensitive adult sensory neurons. Using the whole-cell patch-clamp recording technique, exposure of isolated sensory neurons to either 100 ng/mL NGF or 1 mmol/L N-acetyl sphingosine (C2-ceramide) produced a 3-4 fold increase in the number of action potentials (APs) evoked by a ramp of depolarizing current in a time-dependent manner. Intracellular perfusion with bacterial sphingomyelinase (SMase) also increased the number of APs suggesting that the release of native ceramide enhanced neuronal excitability. Glutathione, an inhibitor of neutral SMase, completely blocked the NGF-induced augmentation of AP firing, whereas dithiothreitol, an inhibitor of acidic SMase, was without effect. In the presence of glutathione and NGF, exogenous ceramide still enhanced the number of evoked APs, indicating that the sensitizing action of ceramide was downstream of NGF. To investigate the mechanisms of actions for NGF and ceramide, isolated membrane currents were examined. Both NGF and ceramide facilitated the peak amplitude of the TTX-resistant sodium current (TTX-R I(Na)) by approximately 1.5-fold and shifted the activation to more hyperpolarized voltages. In addition, NGF and ceramide suppressed an outward potassium current (I(K)) by ~35%. The inflammatory prostaglandin, PGE2, produced an additional suppression of I(K) after exposure to ceramide (~35%), suggesting that these agents might act on different targets. Based on the existing literature, it is not clear whether this NGF-induced sensitization is mediated by the high-affinity TrkA receptor or the low-affinity p75 neurotrophin receptor. Pretreatment with the p75 blocking antibody completely prevents the NGF-induced increase in the number of APs evoked by the current ramp. Although the sensitization by NGF was blocked, the antibody had no effect on the capacity of ceramide, a putative downstream signalling molecule, to enhance the excitability. Ceramide can be metabolized by ceramidase to sphingosine (Sph) and Sph to sphingosine 1-phosphate (S1P) by sphingosine kinase. It is well established that each of these products of sphingomyelin metabolism can act as intracellular signalling molecules. This raises the question as to whether the enhanced excitability produced by NGF was mediated directly by ceramide or required additional metabolism to Sph and/or S1P. Sph applied externally did not affect the neuronal excitability whereas internally perfused Sph augmented the number of APs evoked by the depolarizing ramp. Furthermore, internally perfused S1P enhanced the number of evoked APs. This sensitizing action of NGF, ceramide, and internally perfused Sph, were abolished by dimethylsphingosine (DMS), an inhibitor of sphingosine kinase. In contrast, internally perfused S1P enhanced the number of evoked APs in the presence of DMS. These observations support the idea that the metabolism of ceramide/Sph to S1P is critical for the sphingolipid-induced modulation of excitability. Thus, our findings indicate that the pro-inflammatory agent, NGF, can rapidly enhance the excitability of sensory neurons. This NGF-induced sensitization is mediated by activation of the sphingomyelin signalling pathway wherein intracellular S1P derived from ceramide, acts as an internal second messenger to regulate membrane excitability, however, the effector system whereby S1P modulates excitability remains undetermined.

Functional significance of lipid changes under the effect of inhalational anesthetics

Small concentrations of inhaled anesthetics can induce type II pneumocytes dysfunction and affect surfactant production and exacerbate oxidant mediated lung injury. The aim of the present work was to study the interrelationship between alveolar surfactant lipid composition and that of the lung tissue aiming to understand the sequence of events in the pathogenesis and pathophysiology of acute lung injury [ALI] after inhalation anesthesia. 60 albino rats were used in this study. Lipids were extracted from the lung tissue and alveolar surfactant obtained by bronchoalveolar lavage from rats which were subjected to 1-2% halothane or nitrous oxide anesthetics. The following parameters were determined on the extracted lipids: total lipids [TL], triglycerides [TG], total cholesterol [T-ch], total free fatty acids [FFA], total phospholipids [TPL], phospholipids fractions, lecithin [phosphatidyl cholin Pc], lysolecithin, cephalin [phosphatidyl ethanol amine PI] and sphingomyelin. I- Effect of halothane In the surfactant: Halothane decreased all parameters studied with the exception of FFA and sphingomyelin which was increased compared to controls. Cephalin did not change. However, in the lung FFA, lecithin and lysolecithin increased while all other parameters decreased. II- Effect of nitrous oxide [N[2]O]: In the surfactant: All parameters also decreased except lysolecithin, cephalin and sphingomyelin, which were increased. However, FFA did not change significantly from controls. In the lung: Lecithin, lysolecithin and FFA increased whereas the other parameters decreased. Halothane and N[2]O differ as regards their effect on the lipid profile of the surfactant or lung tissue. The functional significance of the alteration was discussed

[Niemann pick type A, a case report]

Niemann Pick type A is a very rare hereditary disease with an incidence 1 in 20000-40000 live birth, which is calassified as a shingolipidoses. The disease is marked by the abnormal accumulation of sphingomyelin in most tissues, secondary to sphingomylinase deficiency. The most clinical manifestations are: Splenohepatomegaly-cherry red maculae-neuropathologic findings. This is a case report of an infant with clinical manifestation of Niemann Pick disease type A

Surface activity, lipid profiles and their implications in cervical cancer.

BACKGROUND: The profiles of lipids in normal and cancerous tissues may differ revealing information about cancer development and progression. Lipids being surface active, changes in lipid profiles can manifest as altered surface activity profiles. Langmuir monolayers offer a convenient model for evaluating surface activity of biological membranes. AIMS: The aims of this study were to quantify phospholipids and their effects on surface activity of normal and cancerous human cervical tissues as well as to evaluate the role of phosphatidylcholine (PC) and sphingomyelin (SM) in cervical cancer using Langmuir monolayers. METHODS AND MATERIALS: Lipid quantification was done using thin layer chromatography and phosphorus assay. Surface activity was evaluated using Langmuir monolayers. Monolayers were formed on the surface of deionized water by spreading tissue organic phase corresponding to 1 mg of tissue and studying their surface pressure-area isotherms at body temperature. The PC and SM contents of cancerous human cervical tissues were higher than those of the normal human cervical tissues. Role of PC and SM were evaluated by adding varying amounts of these lipids to normal cervical pooled organic phase. Statistical analysis: Student's t-test (p < 0.05) and one-way analysis of variance (ANOVA) was used. RESULTS: Our results reveals that the phosphatidylglycerol level in cancerous cervical tissue was nearly five folds higher than that in normal cervical tissue. Also PC and sphingomyelin SM were found to be the major phospholipid components in cancerous and normal cervical tissues respectively. The addition of either 1.5 microg DPPC or 0.5 microg SM /mg of tissue to the normal organic phase changed its surface activity profile to that of the cancerous tissues. Statistically significant surface activity parameters showed that PC and SM have remarkable roles in shifting the normal cervical lipophilic surface activity towards that of cancerous lipophilic component. CONCLUSION: The Langmuir monolayer technique was sensitive to detect changes in tensiometric profiles of cervical cancers and these could be modulated by alterations in phosphatidylcholine and sphingomyelin levels. Therapeutic strategies may be designed to modulate these tensiometric profiles and lipid constituents of cancerous tissues.

Correlación del recuento de cuerpos laminares con el índice de lecitina/esfingomielina y/o la presencia de fosfadilglicerol para la determinación de madurez pulmonar fetal / Correlation of the lamellar bodies count with lecithin/sphingomyelin ratio and/or the presence of phosphadylglicerol for determination of fetal lung maturity

El objetivo del estudio fue determinar si el recuento de Cuerpos Laminares mayor a 30.000 por uL se correlaciona con pruebas "Gold Standard" cuando estas indican que existe madurez pulmonar fetal (38, 39, 41). Se correlacionó el recuento obtenido en 22 muestras de líquido amniótico de amniocentesis realizadas en el primer semestre del año 2.000, con 2 pruebas de referencia, como son el índice de leticina/Esfingomielina > 2 (19, 22) y/o la presencia de fosfalidilglicerol (20, 26), El recuento de cuerpos laminares se efectuó en un contador de plaquetas Celldyn 1400 de Abbott y las muestras debieron cumplir con los requisitos de nuestro protocolo, los cuales fueron: embarazos con edad gestacional entre 32 a 36 semanas que requirieron amniocentesis por alguna indicación médica y embarazadas con diabetes insulino requirientes de 32 a 40 semanas a quienes se las realizó amniocentesis antes de su parto; además que las muestras no estuvieran contaminadas con sangre ni meconio. Los resultados demostraron que un recuento mayor a 30.000 cuerpos laminares por uL, (12 pacientes), se correlacionaron en un 100 por ciento con una o ambas pruebas de referencia cuando estas indican madurez pulmonar fetal. El recuento de cuerpos laminares es de muy bajo costo (33, 34, 41) y técnicamente accesible en los servicios públicos ya que solo se requiere de un equipo para el recuento de plaquetas, pudiendo ser utilizada para determinar madurez pulmonar fetal cuando otras pruebas clásicas como el Test de Clements dan resultado negativo (porcentaje de falsos inmaduros elevado)

Manifestaçäo hemorrágica no recém nascido como sinal precoce da doença de Niemann-Pick / Bleeding in newborn as early signal of Niemann-Pick

A Doença de Niemann-Pick é um distúrbio de armazenamento hereditário cujo problema principal reside na deficiência parcial ou total de esfingomielinase e/ou de suas isoenzimas, levando ao acúmulo de esfingomielina em todos os orgäos, principalmente no fígado, rins e cérebro. As alteraçöes da coagulaçäo na Doença de Niemann-Pick ainda säo pouco descritas, porém podem apresentar-se precose e isoladamente nos neonatos, precedendo os sinais e sintomas mais frequentes. Os autores apresentam neste relato um caso de paciente do sexo feminino, cinco meses de vida, com quadro de hemorragia grave aos 13 dias de vida após realizaçäo do Teste do Pezinho e aumento do volume abdominal progressivo desde o nascimento. O diagnóstico de Doença de Niemann-Pick foi dado após exclusäo das principais patologias com quadro semelhante e pelo achado de células histiocíticas com depósito citoplasmático espumoso no aspirado de medula óssea. Os autores destacam ainda suas características clínico-laboratoriais e diagnósticos diferenciais.

Avaliação da maturidade pulmonar fetal em gestaçöes de alto risco / Prenatal diagnosis of fetal lung maturity in high-risk pregnancies

Trata-se de um estudo prospectivo para a avaliaçao da maturidade fetal em 121 gestaçoes de alto risco realizado no Hospital Sao Paulo - Universidade Federal de Sao Paulo, entre janeiro de 1990 e janeiro de 1995. Em todos os casos, o parto foi realizado em até 3 dias após a obtençao de líquido amniótico por amniocentese. O objetivo principal foi o de analisar a acurácia do teste de Clements (TC), da relaçao lecitina/esfingomielina (L/E), da presença de fosfatidilglicerol (PG) e do perfil pulmonar (relaçao L/E>1,7 e PG presente) para antecipar a ocorrência ou nao de síndrome de desconforto respiratório neonatal (SDR). Foram calculados a sensibilidade, a especificidade e os valores preditivos positivo (VPP) e negativo (VPN) de todos os testes. O grupo de estudo foi composto por 48 geraçoes complicadas por diabetes mellitus, 41 por síndromes hipertensivas, 14 por isoimunizaçao Rh e 18 por diversas patologias. O perfil pulmonar apresentou sensibilidade de 100 por cento em todos os casos. O teste de Clements também nao apresentou resultados falso-positivos em gestantes hipertensas apurando-se, contudo, de 20 por cento a 50 por cento de falso-negativos em todos os outros testes. Os quatro testes apresentaram baixos VPP (23 por cento no TC, 51 por cento na relaçao L/E, 63 por cento na presença de PG, 61 por cento no perfil pulmonar) e elevados VPN (92 por cento no TC, 88 por cento na relaçao L/E, 89 por cento na presença de PG, 100 por cento no perfil pulmonar). Este estudo demonstrou que a presença de PG e relaçao L/E>1,7 simultâneos no líquido amniótico comprovam a maturidade pulmonar com muito baixo risco de Dr ao nascimento. Concluiu-se também que o teste de Clements deve constituir o rastreamento inicial para predizer a ausência de SDR, particularmente em gestaçoes complicadas por síndromes hipertensivas.

The relationship of sonographic placental grading, biochemical pulmonary maturity and neonatal outcome in pregnancy-induced hypertension.

In a prospective study, the accuracy of placental grade in predicting pulmonary maturity was evaluated in 50 cases of pregnancy-induced hypertension (PIH) between 28 and 41 weeks of gestation. Pulmonary maturity was measured by lecithin/sphingomyelin (L/S) ratio and phosphatidylglycerol (PG) in amniotic fluid and by clinical development of respiratory distress syndrome (RDS) in the neonates. Fifty normotensive healthy primigravidae, who were matched for age and period of gestation, served as control. No difference (p > 0.05) in the placental grading was observed between normotensive and hypertensive pregnancies. The advancement of placental grade was found to be associated with an increase in L/S ratio and PG level in amniotic fluid. A mature placental grade (grade III) identified by real-time sonography corresponded to foetal lung maturity (L/S > or = 2.0, PG > or = 0.36 mg/dl) and absence of RDS in all cases. Hence, in PIH a grade III placenta appeared to be a reliable predictor of foetal lung maturity in the population examined.

Niemann-Pick disease type B with oculo-neural involvement

Niemann-Pick disease type B was diagnosed clinically and enzymatically in a 1-year-old Saudi boy presenting with hepatosplenomegaly, failure to thrive and foam cells in the bone marrow aspirate. Neurological examination and EEG were normal. Sphingomyelinase activity was deficient in leukocytes and cultured skin fibroblasts. Fundoscopy revealed "cherry red spots" of both maculas. By definition, patients with Niemann-Pick disease type B should have no cerebral involvement, and rare ocular involvement [the maculahalosyndrome], which has been described as a pathognomonic sign of this rare disease. For classification - and especially for genetic counseling - it seems important to include oculo-neural involvement in the diagnosis of Niemann-Pick disease type B

Estudo comparativo entre espectrofotometria do líquido amniótico (DDO 650) e a relaçåo lecitina/esfingomielina / A comparative study between amniotic fluid spectrophotometry and lecitin/sphingomelin ratio

Foi realizada uma comparaçåo entre a espectrofotometria (DDO 650) do líquido amniótico e a relaçåo L/E. Encontrou-se significativa correlaçåo entre o resultado da espectrofotometria (DDO 650) igual ou acima de 0,150 e relaçåo L/E igual ou maior que dois. Os autores sugerem a possibilidade de uso do estudo espectrofotométrico pela sua simplicidade, baixo custo e rapidez de execuçåo

Enfermedad de niemann pick error innato del metabolismo de los lípidos: reporte de caso

Se hace una revisión de la enfermedad de Niemann-Pick que es una alteración innata del metabolismo de los fosfolípidos, la ESPINGOMIELINA, que se acumula en los lisosomas, constituyendo la enfermedad lisosomal, debido a una alteración cualitativa o cuantitativa de la ESZPINGOMIELINASA. La enfermedad es transmitida en forma autosómica recesiva, alterando los diversos órganos, especialmente el sistema nervioso y el fondo de ojo en el cual se observa en la mácula una mancha de color cereza. A continuación se presenta el caso problema en un indígena puro del Cañar-Ecuador (años anteriores se encontró otro caso que fue descrito por su extrema rareza), existe consanguinidad; hepatoesplenomegalia, mancha roja en mácula y células espúmosas en médula ósea. Un hermano falleció con identicas características clínicas.