Clinical usefulness of prognostic biomarkers in optic neuritis.

BACKGROUND AND PURPOSE: Different biological and radiological biomarkers predict clinical conversion to multiple sclerosis (MS) after a clinically isolated syndrome (CIS). The aim was to explore their role in predicting the outcome of patients with optic neuritis (ON), a CIS considered to have a benign prognosis. METHODS: Sixty-eight consecutive ON patients were followed prospectively. Magnetic resonance imaging (MRI) and cerebrospinal fluid studies including oligoclonal immunoglobulin G (IgG) bands (OCGBs), lipid-specific oligoclonal IgM bands (LS-OCMBs) and neurofilament light chain quantification were performed at disease onset. Conversion to clinically definite MS (CDMS) was monitored. RESULTS: The mean time of follow-up of our series was 46.4 months. Twenty-five patients (36.7%) developed CDMS during follow-up. Neurofilament light chain levels did not predict clinical conversion. By contrast, an abnormal MRI increased the risk of CDMS [hazard ratio (HR) 12.5, P = 0.013]. The clearest association was found in patients with more than three T2 lesions. OCGBs also predicted the onset of CDMS (HR 21.3, P = 0.003) and LS-OCMBs were associated with a shorter time to CDMS (HR = 116.6, P < 0.001). CONCLUSIONS: Magnetic resonance imaging and OCGBs predicted conversion to CDMS after an ON episode. In addition, LS-OCMBs identified the ON patients more likely to develop MS early. These results, applicable to the everyday clinical setting, may be of interest for therapeutic decisions.

Natural killer cell subsets in cerebrospinal fluid of patients with multiple sclerosis.

Changes in blood natural killer (NK) cells, important players of the immune innate system, have been described in multiple sclerosis (MS). We studied percentages and total cell counts of different effector and regulatory NK cells in cerebrospinal fluid (CSF) of MS patients and other neurological diseases to gain clearer knowledge of the role of these cells in neuroinflammation. NK cell subsets were assessed by flow cytometry in CSF of 85 consecutive MS patients (33 with active disease and 52 with stable MS), 16 with other inflammatory diseases of the central nervous system (IND) and 17 with non-inflammatory neurological diseases (NIND). MS patients showed a decrease in percentages of different CSF NK subpopulations compared to the NIND group. However, absolute cell counts showed a significant increase of all NK subsets in MS and IND patients, revealing that the decrease in percentages does not reflect a real reduction of these immune cells. Remarkably, MS patients showed a significant increase of regulatory/effector (CD56(bright) /CD56(dim) ) NK ratio compared to IND and NIND groups. In addition, MS activity associated with an expansion of NK T cells. These data show that NK cell subsets do not increase uniformly in all inflammatory neurological disease and suggest strongly that regulatory CD56(bright) and NK T cells may arise in CSF of MS patients as an attempt to counteract the CNS immune activation characteristic of the disease.

Cerebrospinal fluid immunological biomarkers associated with axonal damage in multiple sclerosis.

BACKGROUND AND PURPOSE: Cerebrospinal fluid (CSF) neurofilament light protein (NFL) is a promising biomarker of axonal injury and neurodegeneration. Here CSF lymphocyte subpopulations and antibodies, potential players of neurodegeneration, are examined in relation to CSF NFL shedding in MS. METHODS: Cerebrospinal fluid NFL from 127 consecutive untreated MS patients was analysed. Samples from 37 age-matched patients with other central nervous system non-inflammatory neurological diseases (NIND) were also assessed. CD4+, CD8+, CD56+ and CD19+ cell subsets were studied by flow cytometry. Oligoclonal IgG and IgM bands (OCMB) against lipids were studied by isoelectric focusing and immunoblotting. These data were analysed in relation to clinical and magnetic resonance imaging features. RESULTS: A CSF NFL cut-off value of 900 ng/l (mean + 3 SD of NIND values) was calculated. MS patients with increased NFL values showed significantly higher Multiple Sclerosis Severity Score and magnetic resonance imaging lesion number. The presence of OCMB (P < 0.0001) and elevated T and B lymphocyte counts was associated with increased levels of CSF NFL. CONCLUSIONS: High CSF NFL levels are associated with elevated CSF lymphocyte cell counts and intrathecal synthesis of IgM against lipids. These findings support a role for OCMB in the axonal damage of MS offering a rationale for the association of these antibodies with disability and brain atrophy progression in MS.

Nitrogen modulates the ozone response of Mediterranean wheat: Considerations for ozone risk assessment.

The experiment was conducted in an Open Top Chamber facility located in the Mediterranean basin to investigate how nitrogen (N) fertilization affects the response of wheat to ozone (O3) exposure. The study considered the response of Artur Nick, a modern wheat cultivar commonly used in the area, to three O3 exposure levels (ambient and elevated ambient, +20 and +40 nL L-1 O3), and two N fertilization doses (100 and 200 kg ha-1). Measurements included leaf gas exchange, leaf chlorophyll content, leaf and grain N content, plant growth and yield parameters. Ozone × N interactive effects were studied and quantified based on accumulated O3 concentrations above a 40 nL L-1 threshold (AOT40) and phytotoxic O3 dose (POD) indices, which are used in O3-risk assessments, from which critical levels (CL) for a 5 % effect were derived. Results revealed that O3 impacts on growth and yield parameters were stronger under the highest N fertilization dose. In consequence, O3 Critical Levels (CL) were as much as 3-4 times lower for grain yield in the high-N compared to the low-N treatment. Interestingly, O3 limited the fertilizer stimulus, strongly reducing the N use efficiency for grain yield and the agronomic efficiency of N for protein yield. Another important aspect was that 71 % of the POD was accumulated before anthesis, stressing the potential importance of O3 exposure during the vegetative phase of wheat under Mediterranean conditions, which is usually considered less important than post-anthesis exposure. In conclusion, this study suggests the need to consider crop N management in the derivation of O3 CLs, due to its effect on dose-response relationships used for CL derivation, including the potential O3 effects in N use efficiency. Therefore, N modulation could be considered in the O3-risk assessment methodology to be applied in risk exercises when negotiating air pollution abatement policies.

Current ambient ozone levels mitigate the effect of Puccinia striiformis on wheat: Is Mediterranean wheat ready for pre-industrial background ozone levels?

Increasing surface ozone is a main concern for crop production in the Global Change framework, especially in the Mediterranean basin where climate conditions favor its photochemical formation. Meanwhile, increasing common crop diseases, such as yellow rust, one of the most important pathogens affecting global wheat production has been detected in the area in recent decades. However, the impact of O3 on the occurrence and impact of fungal diseases is scarcely understood. A close-to-field-conditions assay (Open Top Chamber facility) situated in a Mediterranean cereal rainfed farming area was carried out to study the impact of increasing O3 levels and N-fertilization on spontaneous fungal outbreaks in wheat. Four O3-fumigation levels reproducing pre-industrial to future pollutant atmospheres with additional 20 and 40 nL L-1 over the ambient levels were considered (7 h-mean ranging from 28 to 86 nL L-1). Two top N-fertilization supplementations (100 and 200 kg ha-1) were nested within the O3 treatments; foliar damage, pigment content and gas exchange parameters were measured. Pre-industrial natural background O3 levels strongly favored the yellow rust infection, where the O3-polluted levels currently observed at the farm highly benefited the crop, mitigating the presence of rust by 22 %. However, future expected high O3-levels neutralized the beneficial infection-controlling effect by inducing early wheat senescence, decreasing the chlorophyll index of the older leaves by up to 43 % under the higher O3 exposure. Nitrogen promoted the rust infection by up to 49.5 % without interacting with the O3-factor. Achieving future air quality standards might require considering new varietal improvement programs, to be able to adapt crops to an increased pathogen tolerance without requiring the assistance provided by O3-pollution.

Multiple sclerosis patients with anti-lipid oligoclonal IgM show early favourable response to immunomodulatory treatment.

BACKGROUND AND PURPOSE: Interferon beta and Glatiramer acetate are safe immunomodulatory treatments (IT) for multiple sclerosis (MS), but not always effective. New drugs are available, although they show more side-effects and unknown long-term safety profile. Anti-lipid oligoclonal IgM bands (OCMB) distinguish MS patients with early aggressive course. We prospectively studied if IT are effective in these patients or if they are candidates for more aggressive drugs as first therapeutic option. METHODS: Seventy-five clinically isolated syndrome patients were studied. OCMB and conversion to MS were assessed. Patients suffering at least two demyelinating events within 3 years were considered eligible to start IT. RESULTS: Eighteen patients showed OCMB (M+) and 57 lacked them (M-). All M+ patients and only 25 M- patients were treated. The other 32 M- patients suffered less MS attacks than those required to initiate treatment. IT similarly reduced relapse rate in both treated groups (P < 0.0001) and reduced Expanded Disability Status Scale (EDSS) progression in M+ patients, whose EDSS score had significantly increased before treatment. EDSS did not change in M- patients during follow-up, regardless if they were treated or not. CONCLUSIONS: Oligoclonal IgM bands identify MS patients who are candidates for early immunomodulatory treatment as IT improves their initial aggressive disease course.

Simulating the migration of drifters deployed in the Bay of Biscay, during the Prestige crisis.

A main conclusion following the oil spill from the Prestige tanker was that improvements in ocean circulation models were necessary; this was in order to predict, more accurately, the trajectories followed by the oil slicks and hence assist in fight against oil pollution operations. In this contribution, the results of the validation of a semi-empirical ocean circulation model, parameterised for the Bay of Biscay and forced with operational oceano-meteorological remote sensing observations, are shown. The model results have been validated with observations from drifting buoys, deployed in the Bay of Biscay during the crisis. The results show that the model explains a relatively large percentage of the current variability. The comparisons between the real and the estimated drifter trajectories indicate that for 3, 5 and 7 day-long trajectories, the drifter position is estimated with errors of approximately 23, 35 and 46km, respectively. The model reproduces relatively well the trajectory followed by the drifter with the shortest period (23 days).

Multiple myeloma patients in long-term complete response after autologous stem cell transplantation express a particular immune signature with potential prognostic implication.

The proportion of multiple myeloma patients in long-term complete response (LTCR-MM) for more than 6 years after autologous stem cell transplantation (ASCT) is small. To evaluate whether this LTCR is associated with a particular immune signature, peripheral blood samples from 13 LTCR-MM after ASCT and healthy blood donors (HBD) were analysed. Subpopulations of T-cells (naïve, effector, central memory and regulatory), B-cells (naïve, marginal zone-like, class-switched memory, transitional and plasmablasts) and NK-cells expressing inhibitory and activating receptors were quantified by multiparametric flow cytometry (MFC). Heavy/light chains (HLC) were quantified by nephelometry. The percentage of CD4+ T-cells was lower in patients, whereas an increment in the percentage of CD4+ and CD8+ effector memory T-cells was associated with the LTCR. Regulatory T-cells and NK-cells were similar in both groups but a particular redistribution of inhibitory and activating receptors in NK-cells were found in patients. Regarding B-cells, an increase in naïve cells and a corresponding reduction in marginal zone-like and class-switched memory B-cells was observed. The HLC values were normal. Our results suggest that LTCR-MM patients express a particular immune signature, which probably reflects a 'high quality' immune reconstitution that could exert a competent anti-tumor immunological surveillance along with a recovery of the humoral immunity.

Laboratory guidelines for the diagnosis and follow-up of patients with monoclonal gammopathies.

We present guidelines from the Immunochemistry group of the Spanish Society for Immunology that are designed to provide a practical tool for the diagnosis and follow-up of monoclonal gammopathies. We review the clinical and analytical features of various monoclonal gammopathies, international consensus guidelines and techniques used to detect and follow-up monoclonal components.

Assessment of the reproducibility of oligoclonal IgM band detection for its application in daily clinical practice.

BACKGROUND: The presence of oligoclonal IgM bands (OCMB) in cerebrospinal fluid (CSF) is an unfavourable prognostic marker in multiple sclerosis. There is no commercial test to investigate OCMB status. However, a sensitive and specific isoelectrofocusing (IEF) and western blot method was described. We aimed to study the inter-centre reproducibility of this technique, a necessary condition for a reliable test to be incorporated into clinical practice. METHODS: The presence of OCMB was analysed by IEF and western blot with prior reduction of pentameric IgM. We assayed the reproducibility of this test in a blinded multicentre study performed in 13 university hospitals. Paired-CSF and serum samples from 52 neurological patients were assayed at every centre. RESULTS: Global analysis rendered a concordance of 89.8% with a kappa value of 0.71. CONCLUSION: These data indicate that OCMB detection by means of IEF and western blot with IgM reduction shows a good interlaboratory reproducibility and thus can be used in daily clinical setting.

Effect of Depo-Provera or Microgynon on the painful crises of sickle cell anemia patients.

Forty-three homozygous (SS) female sickle cell anemic patients with a history of at least one painful crisis per month and desiring a reversible contraceptive were administered DMPA/3 months or Microgynon monthly. A third group of 16 surgically sterilized patients served as control. Patients were followed for 1 year to assess possible effects of the contraceptives on the patients' painful crises. No changes were observed in any of the groups in the hematological parameters. At the end of the study, 70% of the patients receiving DMPA were pain-free and only 16% of those still reporting painful crises rated them as intense. Patients receiving Microgynon also had an amelioration of the painful crises, although at a lower rate; after 12 months, 45.5% still experienced some crises. Although less marked than in the other groups, 50.5% of the control patients also reported an improvement of their painful crisis, which may be a result of closer medical care.

PIP: The effects of a combined oral contraceptive (Microgynon 30, containing ethinyl estradiol and levonorgestrel) and a progestogen-only injectable contraceptive (Depo-Provera) on the intensity and frequency of painful crises were investigated in 43 homozygous sickle cell anemia patients at the World Health Organization Collaborative Center for Research in Human Reproduction in Panama. Only women with a history of at least one painful crisis per month were enrolled. The patients were randomly assigned to receive Depo-Provera (n = 13) or Microgynon (n = 14) for 12 months; the remaining 16 patients--surgically sterilized controls-- received no treatment. No changes were recorded throughout the study period in any of the three groups in hematological parameters. In addition, there were no pregnancies or treatment-related side effects. In the Depo-Provera group, the percentage of patients with painful crises diminished steadily from 50% at 3 months to 30% at 12 months. Moreover, 84% of painful episodes experienced by Depo-Provera acceptors were characterized as moderate or mild. Among women in the Microgynon group, the rate of painful crises dropped from 72.7% at 3 months to 45.5% at 12 months. Controls also reported a 50.5% decline in painful crises, presumably as a result of increased individual attention from medical staff. These findings suggest that Depo-Provera can be safely and effectively used for contraceptive purposes in sickle cell anemic patients, with a concomitant beneficial effect on painful crises.

A spectrum in the pathology of toxoplasmosis in patients with acquired immunodeficiency syndrome.

We describe a variety of toxoplasmic lesions in seven patients with the acquired immunodeficiency syndrome. The first patient had multiple small-intestinal ulcers associated with Toxoplasma tachyzoites and high antibody titers; he died of disseminated histoplasmosis. The second patient, who died of tuberculosis, also had an inactive chronic Toxoplasma infection, with tissue cysts in the brain that were associated with glial nodules. A third patient died of Toxoplasma encephalitis, manifested by multiple foci of necrosis associated with Toxoplasma tachyzoites, cysts, and hypertrophic arteritis. A fourth patient had been treated for toxoplasmic encephalitis with co-trimoxozol (trimethoprim-sulfamethoxazole combination) for 3 to 4 days and showed degenerating tachyzoites associated with necrotic areas. A fifth patient, treated for toxoplasmic encephalitis with co-trimoxazol for 14 days, had necrotic lesions associated with Toxoplasma antigen and a few cysts. A sixth patient with encephalitis and Toxoplasma tachyzoites and young cysts in the biopsy showed healed brain lesions after 22 days of treatment. A seventh patient, diagnosed radiologically and serologically with Toxoplasma encephalitis, was treated for 7 months; his ring-enhancing lesions subsided, and he died of a central nervous system lymphoma. Toxoplasma could not be isolated from the brain, although toxoplasmic DNA was detected in the brain and heart by polymerase chain reaction. The pathogenesis of the range of these lesions, their diagnosis, and the possibility of terminating Toxoplasma infection by prolonged chemotherapy are discussed.

Chihuahua: a water reuse case in the desert.

Water supply for all kind of uses in Chihuahua is mainly groundwater. During the last decade this city has been damaged with a heavy hydrologic crisis because of a persistent drought. This came up with the overexploitation of groundwater aquifers; therefore a deficit between demand and offer was done. To minimize this problem the government authorities have started an integral plan of optimizing hydrologic resources which considers the treatment of wastewater and the use of reclaimed water. The secondary wastewater treatment facility of the city treats about 30,000 m3/d of a wastewater with high organic contents, and produces an effluent with low concentration of suspended solids, organic matter, fats, detergents, and metals. Reclaimed water is conveyed toward strategic sites for the irrigation of great green areas in sport clubs, educational institutions and industrial zones, besides of its utilization on some manufacturing processes, road service, and also over construction industry. The potential reuse of this water goes farther from those activities; the treatment of the secondary effluent until the required levels of the water-bearing recharge criteria are met for drinking water supply is considered as the next step to achieve through a suitable planning strategy for the best integral resource advantage.

[Pseudotumor cerebri associated with maxillary sinusitis: clinico-pathological study]. / Pseudotumor cerebral asociado a sinusitis maxilar y pleocitosis en líquido cefalorraquídeo.

INTRODUCTION: The pseudotumor cerebri is characterized by increased intracranial pressure in the absence of a space occupying lesions or obstruction to the circulation of cerebrospinal fluid (CSF). Increased pressure but normal composition of the CSF is the diagnostic criterion. Ear infections and other infections such as sinusitis have been cited as possible, but rare, causes. CLINICAL CASE: A 12 year old girl presented with diplopia 10 days after starting treatment for sinusitis. On examination there was paresia of the VI cranial nerve to the left eye and marked bilateral papilledema. She had normal visual acuity with visual field measurements showing increased blind spot field in both eyes. Resonance studies only showed maxillary sinusitis. CSF pressure was 35 cm H2O, with 40 lymphomonocytes and normal biochemical findings. After 15 cm of CSF had been removed treatment was started with acetazolamide, dexamethasone and cefotaxime. A week later the diplopia had disappeared and the CSF pressure was 25, with no cells present. Two months later the visual fields and fundus oculi were normal. After follow-up for one year there was no recurrence. CONCLUSIONS: We describe a case of pseudotumor cerebri associated with maxillary sinusitis. We do not know whether this is a chance association or whether there was a pathophysiological basis. The possibility should be considered so as to treat the cause or predisposing factor. The initial CSF pleocytosis would support the possibility of a relationship of the pseudotumor with an infectious condition (meningeal irritation or para-infectious pleocytosis meningeal irritation or para-infections pleocytosis in the context of an ENT infection), which might have precipitated the problem by interfering with the reabsorption of CSF.

[DDT and related pesticides in maternal milk and other tissues of healthy women at term pregnancy]. / DDT y plaguicidas relacionados presentes en la leche materna y otros tejidos de mujeres sanas con embarazos de término.

Ten healthy patients with term pregnancy resolved by abdominal via. During the surgical procedure samples of umbilical chord serum and maternal adipose tissue, were taken. In all the samples, together with the maternal milk collected the day 10 of puerperium, the concentrations of the following organochlorine pesticides, were measured up: (PCC); beta-BHC; gamma-BHC; heptachloride; aldrin; dieldrin, DDE, DDD, DDT and methoxychloride. The identification and quantification of pesticides was done by the comparison with standards certified by NIST (National Institute of Standard Technology). The general characteristics of the participants were: primigestas of 24.1 years aged, married and of a low socioeconomical level. As to the neonates, of 39 weeks of gestational age, female sex; 3,311 g of corporal weight and size of 51.1 cm. In all the analyzed samples at least one of the organochlorine pesticides was present. The results of correlation analysis between DDT concentration, present in the maternal serum with those identified in the adipose tissue and serum from the umbilical chord were highly significant: a = 0.97 and 0.87, respectively. In the maternal milk the highest concentrations of total DDT, were found, average of 2053 ng/g lipidic base, which is 2.8 times more of daily accepted intake. Likewise, DDT concentration in maternal serum kept exponential relation, growing with age (a = 0.99).

Influence of oligoclonal IgM specificity in multiple sclerosis disease course.

Oligoclonal IgM bands (OCMB) against myelin lipids predict an aggressive multiple sclerosis (MS) course. However, the clinical significance of OCMB without lipid specificity, present in other MS patients, remains unknown. We describe here a characterization of these antibodies and study their role in MS progression. Fifty-four MS patients showing CSF-restricted OCMB were included in this study at disease onset and followed-up during 61.1 +/- 2.7 months. The specificity of OCMB and the CSF B-cell profile were investigated. A second CSF IgM study was performed in a group of eight patients. Thirty-eight patients showed OCMB against myelin lipids (M+L+) and other sixteen had OCMB lacking this specificity (M+L-). The CD5+ B cell subpopulation, responsible for most persistent IgM responses, was considerably higher in M+L+ than in M+L- patients (3.3 +/- 0.6% versus 0.8 +/- 0.2, P = 0.009). In addition, M+L+ bands persisted during disease course, while M+L- disappeared during follow-up. M+L+ patients suffered more relapses (4.2 +/- 0.6 versus 1.6 +/- 0.3, P = 0.002) and reached higher disability (EDSS score of 2.2 +/- 0.2 versus 1.2 +/- 0.2, P = 0.02) than M+L- group. These data corroborate that anti-lipid OCMB associate with an aggressive MS course and show that OCMB that do not recognize myelin lipids represent a transient immune response related to a more benign disease course.

Adipose tissue and liver lipid metabolism in obese children: role of the body mass index and the presence of acanthosis nigricans.

AIMS: The aims of our study were to determine if insulin resistance is associated with increased plasma levels of non-esterified fatty acids (NEFA), glycerol, 3-hydroxybutyrate and triglycerides in obese children. We also studied whether the presence of acanthosis nigricans (AN) led to further alterations in the above parameters. METHODS: A total of 101 children were studied on their first visit to the paediatric endocrine clinic. Seventy-four were obese, 30 of them with AN. The remaining 27 were non-obese healthy children (control group). NEFAs, glycerol, triglycerides, 3-hydroxybutyrate, insulin, leptin, adiponectin and glucose were determined in blood samples obtained after overnight fasting. The insulin resistance index (IRI) was calculated following the homeostasis model assessment (HOMA). Data from the three groups were compared using appropriate statistical tests. RESULTS: No differences in age, sex ratio and pubertal stage were observed among the three groups. The group of children with the highest body mass index (BMI) showed higher plasma levels of insulin and leptin, higher IRI and lower plasma levels of adiponectin. As insulin and IRI increased, NEFA and 3-hydroxybutyrate decreased and triglycerides increased. When obese children were categorized by BMI, the presence of AN further exacerbated these differences. CONCLUSIONS: In obese children, insulin resistance is associated with plasma lipid alterations suggestive of both decreased adipose tissue lipolysis and hepatic beta-oxidation and increased hepatic synthesis of triglycerides. Such a metabolic condition may facilitate fat storage and hinder weight loss.

Clinically isolated syndromes: a new oligoclonal band test accurately predicts conversion to MS.

BACKGROUND: Patients with a clinically isolated demyelinating syndrome (CIS) are at risk of developing a second attack, thus converting into clinically definite multiple sclerosis (CDMS). Therefore, an accurate prognostic marker for that conversion might allow early treatment. Brain MRI and oligoclonal IgG band (OCGB) detection are the most frequent paraclinical tests used in MS diagnosis. A new OCGB test has shown high sensitivity and specificity in differential diagnosis of MS. OBJECTIVE: To evaluate the accuracy of the new OCGB method and of current MRI criteria (MRI-C) to predict conversion of CIS to CDMS. METHODS: Fifty-two patients with CIS were studied with OCGB detection and brain MRI, and followed up for 6 years. The sensitivity and specificity of both methods to predict conversion to CDMS were analyzed. RESULTS: OCGB detection showed a sensitivity of 91.4% and specificity of 94.1%. MRI-C had a sensitivity of 74.23% and specificity of 88.2%. The presence of either OCGB or MRI-C studied simultaneously showed a sensitivity of 97.1% and specificity of 88.2%. CONCLUSIONS: The presence of oligoclonal IgG bands is highly specific and sensitive for early prediction of conversion to multiple sclerosis. MRI criteria have a high specificity but less sensitivity. The simultaneous use of both tests shows high sensitivity and specificity in predicting clinically isolated demyelinating syndrome conversion to clinically definite multiple sclerosis.