Revealing enterovirus infection in chronic human disorders: An integrated diagnostic approach.

Enteroviruses (EVs) causing persisting infection are characterized by minimal replication and genetic changes. Typing of these agents may complement disease assessment and shed light on pathogenesis. Here we report an integrated approach for EV detection in human samples that is based on pre-enrichment of virus in cell culture before search for the viral genome and viral antigens. Cases of post-polio syndrome, type 1 diabetes, and chronic cardiomyopathy were investigated. As tissue-based approaches require invasive procedures, information was mainly gleaned from virus in blood. Molecular assays targeting conserved genome regions of all EV types (5'UTR, 2 C, 3Dpol) were employed. As compared to direct assays of plasma or leukocytes, the EV detection rate was significantly enhanced by co-culture of leukocytes with cell lines prior to molecular and immunologic tests. Results of RT-PCR and sequencing were confirmed by staining cell cultures with a panel of EV-specific antibodies. Sequence and phylogenetic analysis showed that EVs of the C species (polioviruses) were associated with the post-polio syndrome, while members of the B species were found in type 1 diabetes and cardiomyopathy. The procedure may be used for investigating the possible association of different EVs with a variety of chronic neurologic, endocrine, and cardiac disorders.

Elevated plasma inflammatory mediators in post-polio syndrome: No association with long-term functional decline.

A key feature of post-polio syndrome (PPS) is progressive loss of muscle strength. In other chronic diseases systemic inflammation has been linked to muscle wasting. In this study plasma TNF-α, IL-6, IL-8, and leptin levels were significantly increased in PPS-patients compared to healthy controls. There was however no association between these raised systemic levels of inflammatory mediators and long-term decline in quadriceps strength or other clinical parameters. In conclusion, there is evidence for systemic inflammation in PPS, yet the relationship with clinical deterioration remains tenuous.

Elevated blood lipids are uncommon in patients with post-polio syndrome--a cross sectional study.

BACKGROUND: The post-polio syndrome occurs in people who previously have had poliomyelitis. After the initial recovery, new or increasing neurologic symptoms occur. Inflammation and dyslipidaemia may play an important role in the development of atherosclerotic complications, for example myocardial infarction and angina pectoris. Previous studies on cardiovascular risk factors in the post-polio syndrome have found a higher prevalence of hypertension, ischemic heart disease, hyperlipidaemia, and stroke in these patients. The present study was undertaken in order to evaluate whether post-polio patients have elevated lipid values, and if blood lipid abnormalities could be correlated to signs of inflammation. METHODS: Cross-sectional study of 89 consecutive post-polio patients, (53 women, mean age 65 years) from the Post-Polio Outpatient Clinic, Danderyd University Hospital, Stockholm, Sweden. The lipid profiles of post-polio patients were compared to age and sex matched reference values from two earlier studies. Statistical analyses were performed by the Student's t-test, and linear regression analyses were assessed by Pearson's correlation coefficient. RESULTS: Mean total cholesterol levels (5.7 mmol/L) were low or normal in post-polio patients, whereas low density lipoprotein levels (3.6 mmol/L) were normal, and high density lipoprotein (1.5 mmol/L) and triglycerides (1.4 mmol/L) lower than reference values. The prevalence of diabetes (7%), hypertension (38%), concomitant cardiovascular disease, (including angina pectoris, myocardial infarction, heart failure, atrial fibrillation and stroke) (7%), and calculated 10 year risk of coronary heart disease according to Framingham risk score algorithm (8%) was not increased in post-polio patients. CONCLUSIONS: Compared to reference populations, post-polio patients in Sweden appear to have low or normal total cholesterol and low density lipoprotein levels, whereas high density lipoprotein and triglyceride levels are low. Hence, a possible persisting inflammatory process in post-polio syndrome does not seem to be associated with increased lipids and an increased risk for coronary heart disease events.

Genotoxic assessment in peripheral blood lymphocytes of post-polio individuals using sister chromatid exchange analysis and micronucleus assay.

Environmental pollution is a complex issue because of the diversity of anthropogenic agents, both chemical and physical, that have been detected and catalogued. The consequences to biota from exposure to genotoxic agents present an additional problem because of the potential for these agents to produce adverse change at the cellular and organism levels. Past studies in virus have focused on structural damage to the DNA of environmental species that may occur after exposure to genotoxic agents and the use of this information to document exposure and to monitor remediation. In an effort to predict effects at the population, community and ecosystem levels, in the present study, we attempt to characterize damage occurring through genotoxic agents like 5-bromo-2-deoxyuridine, BrdU, using sister chromatid exchange technique and the formation of micronuclei (MN) in the peripheral lymphocytes of the post-polio syndrome sequelae affected by poliovirus. Analysis of structural chromosomal aberrations (CAs) and involvement of the specific chromosome break were pursued in this study. They revealed a significantly higher incidence of CAs (chromatid and chromosome breaks) in patients compared with controls, where the specific chromosome break has emerged as specific. Also, the maximum numbers of breaks were found to be in chromosome 1 at the position 1p36.1. The results also suggest a correlation between CAs and content of MN.

Post-polio syndrome: epidemiologic and prognostic aspects in Brazil.

OBJECTIVES: To describe the clinical and epidemiological aspects of post-polio syndrome (PPS) and identify predictors of its severity. MATERIALS AND METHODS: 132 patients with PPS were selected at the Neuromuscular Disease Outpatient Clinic of the Federal University of São Paulo. Descriptive analysis was carried out and predictors of PPS severe forms were investigated using an unconditional logistic regression. RESULTS: The average age at onset was 39.4 years. The most common symptoms were fatigue (87.1%), muscle pain (82.4%) and joint pain (72.0%); 50.4% of the cases were severe. The following were associated with PPS severity: a < or =4-year period of neurological recovery (OR 2.8), permanent damage in two limbs (OR 3.6) and residence at the time of acute polio in a city with more advanced medical assistance (OR 2.5). CONCLUSIONS: Health professionals should carefully evaluate polio survivors for PPS and be aware of the implications of muscle overuse in the neurological recovery period.

Elevated serum inflammatory markers in post-poliomyelitis syndrome.

OBJECTIVES: To determine (i) whether serum inflammatory markers TNFalpha, IL-1beta. IL-6, and leptin are increased in post-poliomyelitis syndrome (PPS) compared to healthy controls; and (ii) whether an association exists between elevated inflammatory markers and clinical parameters in PPS. The cause of PPS is unknown, but abnormal inflammatory responses have been implicated in several small studies. METHODS: Serum inflammatory markers were measured (by Luminex) in 51 PPS patients and 26 normal controls. Clinical parameters assessed included disease duration, muscle strength (Medical Research Council sumscore), fatigue (Fatigue Severity Scale and Multidimensional Fatigue Inventory), and pain (visual analog scale scores). RESULTS: In PPS, TNFalpha levels, as well as IL-6 and leptin were significantly increased compared to controls (Wilcoxon rank-sum test, p=0.03 for TNFalpha, p=0.03 for IL-6, p=0.01 for leptin). The elevated TNFalpha levels in PPS were associated with increased pain due to illness (Spearman correlation coefficient r=0.36, 95% C.I. 0.09 to 0.57) and specifically, with muscle pain (r=0.38, 95% C.I. 0.11 to 0.59). There were no correlations between inflammatory markers in PPS and joint pain, muscle strength, fatigue, or disease duration. CONCLUSIONS: Serum TNFalpha, IL-6 and leptin levels are abnormally increased in PPS patients. Elevated TNFalpha levels appear to be specifically associated with increased muscle pain.

Prediction of voluntary activation, strength and endurance of elbow flexors in postpolio patients.

To examine the long-term effects of polio, maximal voluntary strength and voluntary activation of elbow flexor muscles of 177 patients from a postpolio clinic were investigated using twitch interpolation. Muscle endurance was studied in 142 patients during 45 min of submaximal exercise, and predictors of impaired muscle performance were investigated. Twenty-nine of 177 patients (16.4%) had impaired voluntary drive to their elbow flexor muscles, but only 16 (9.0%) had markedly reduced elbow flexor strength, despite 74 (41.8%) reporting they were initially affected in their tested limb and 172 (97.2%) patients reporting new generalized symptoms. Seven patients had impaired muscle endurance in the tests of strength and voluntary drive. During the submaximal exercise, 16 patients (11.3%) had impaired peripheral muscle endurance with normal voluntary activation. These results confirm a low incidence of impaired upper-limb muscle performance in postpolio patients, despite many patients having subjective symptoms consistent with postpolio syndrome. There was an increased relative risk for impaired muscle function in those patients with a subjective decrease in strength in the tested limb, a recent decline in activities of daily living in their tested limb, and who used orthotic devices in their tested limb. Monitoring of function in prior-polio patients with impaired muscle performance may be useful, particularly when combined with investigation of other potential contributory factors to the functional impairment.

GM1 antibodies in post-polio syndrome and previous paralytic polio.

We studied the relationship between post-polio syndrome (PPS) and GM1 antibodies, since such antibodies have been associated with PPS and motor neuron disorders. Sera from 144 patients with previous poliomyelitis (105 paralytic, 22 nonparalytic and 17 PPS), 60 with previous Guillain-Barré syndrome, 44 with amyotrophic lateral sclerosis (ALS) and 22 healthy blood donors were analyzed with ELISA for GM1 IgM, IgG and IgA antibodies. GM1 antibodies were present in 14% of the PPS patients, but the prevalence did not differ significantly from that of the other groups. Our study does not support the hypothesis that GM1 antibodies are involved in the pathogenesis of PPS.

Prior poliomyelitis-evidence of cytokine production in the central nervous system.

In order to study the role of a possible inflammatory reaction in the post-polio syndrome (PPS) four key cytokines were determined by means of mRNA expression in mononuclear cells from cerebrospinal fluid (CSF) and peripheral blood of 13 patients. Data were compared with those of samples from eight non-inflammatory control persons. The PPS-patients displayed increased numbers of CSF cells expressing mRNA for TNF-alpha (p<0.02), IFN-gamma (p<0.02), IL-4 (p<0.001) and IL-10 (p<0.05), in comparison to the non-inflammatory controls. As positive controls, samples from patients with Multiple Sclerosis (MS) were examined. We conclude that there is a chronic intra CNS expression of inflammatory cytokines in PPS, in the range of that in MS, a well known neuroinflammatory disease. However, the pathogenic significance of this is unclear.

Serum insulin-like growth factor-I (IGF-I) does not correlate positively with isometric strength, fatigue, and quality of life in post-polio syndrome.

OBJECTIVES AND BACKGROUND: To determine if serum insulin-like growth factor-I (IGF-I) levels are associated with strength, body mass index (BMI), fatigue, or quality of life in post-poliomyelitis syndrome (PPS). PPS is likely due to a distal disintegration of enlarged post-polio motor units as a result of terminal axonal sprouting. Age-related decline in growth hormone and IGF-I (which support terminal axonal sprouts) is proposed as a contributing factor. METHODS: As part of the North American Post-Poliomyelitis Pyridostigmine Study (NAPPS), baseline data on maximum voluntary isometric contraction (MVIC), BMI, subjective fatigue (fatigue severity scale, Hare fatigue symptom scale), health-related quality of life (short form health survey-36; SF-36), and serum IGF-I levels were gathered on 112 PPS patients. Pearson correlation coefficients were calculated to evaluate the association between serum IGF-I and MVIC in 12 muscles, BMI, two fatigue scales, and SF-36 scale scores. RESULTS: There is a significant inverse correlation of IGF-I levels with MVIC in left ankle dorsiflexors (r=-0.30, P<0.01), and left and right knee extensors (r=-0.22, -0.25, P=<0.01, 0.01), but no significant correlations in other muscles. When men and women were evaluated separately, inverse correlations of IGF-I levels with MVIC were found only in men. IGF-I correlated inversely with BMI (r=-0.32, P=0006) and age (r=-0.32, P=0.0005). IGF-I did not correlate with the fatigue or SF-36 scales. CONCLUSIONS: In this exploratory study, we found that contrary to our expectations, IGF-I did not correlate positively with strength. IGF-I correlated negatively with strength in several lower extremity muscles, BMI, and age. IGF-I is likely not an important factor in the pathogenesis of fatigue and in determining quality of life in PPS, but its role on strength should be studied further.

Strength, endurance, and work capacity after muscle strengthening exercise in postpolio subjects.

OBJECTIVE: To determine whether a 12-week home quadriceps muscle strengthening exercise program would increase muscle strength, isometric endurance, and tension time index (TTI) in postpolio syndrome subjects without adversely affecting the surviving motor units or the muscle. DESIGN: A longitudinal study to investigate the effect of a 12-week exercise program on neuromuscular function and electromyographic variables. SETTING: Neuromuscular laboratory of a university hospital. SUBJECTS: Seven subjects were recruited from a cohort of 12 subjects who had participated in a previous exercise study. All subjects had greater than antigravity strength of the quadriceps. Upon completion of a postpolio questionnaire, all acknowledged common postpolio syndrome symptoms such as new fatigue, pain, and weakness; 6 of the 7 acknowledged new strength decline. INTERVENTION: On Mondays and Thursdays subjects performed three sets of four maximal isometric contractions of the quadriceps held for 5 seconds each. On Tuesdays and Fridays subjects performed three sets of 12 dynamic knee extension exercises with ankle weights. MAIN OUTCOME MEASURES: Neuromuscular variables of the quadriceps muscles were measured at the beginning and completion of the exercise program and included: isokinetic peak torque (ISOKPT, at 60 degrees/sec angular velocity) and total work performed of four contractions (ISOKTW), isometric peak torque (MVC), endurance (EDUR, time subject could hold isometric contraction at 40% of the initial MVC), isometric tension time index (TTI, product of endurance time and torque at 40% of MVC), and initial and final ankle weight (WGT, kg) lifted. Electromyographic variables included: fiber density (FD), jitter (MCD), and blocking (BLK) from single fiber assessment and median macro amplitude (MACRO). Serum creatine kinase (CK) was also measured initially and at 4-week intervals throughout the study. RESULTS: The following variables significantly (p < .05) increased: WGT by 47%, ISOKPT, 15%, ISOKTW, 15%; MVC, 36%; EDUR, 21%; TTI, 18%. The following variables did not significantly (p > .05) change: FD, MCD, BLK, MACRO, and CK. CONCLUSIONS: This home exercise program significantly increased strength, endurance, and TTI without apparently adversely affecting the motor units or the muscle, as the EMG and CK variables did not change.

Normal concentrations of serum insulin-like growth factor-1 in late polio.

A recent study of 10 men with postpolio syndrome indicated a low secretion of growth hormone (GH) as reflected by serum insulin-like growth factor-I (IGF-1). Therefore, 87 patients were studied, 17 to 71 years after acute poliomyelitis, of whom 65% reported the occurrence of new or increased weakness (ie, during the last 2 years) in muscles previously affected by polio. Serum IGF-1 concentrations in the patients were compared with those found in a reference population comprising 392 randomly selected individuals. No differences from the reference population values were observed. No correlation was found between IGF-1 concentrations and the severity of the original polio affliction, the recovery status, the need for ambulation aids, or the presence of new symptoms. The results do not indicate a need for GH substitution treatment of patients with postpolio syndrome.

Studies of growth hormone/insulin-like growth factor-I in polio survivors.

In the later years of life, many polio survivors develop post-polio syndrome manifested by progressive muscular weakness. Previous work by the authors showed low IGF-I level in polio survivors compared to age-matched controls. IGF-I concentration tended to be lower in polio survivors who were older, male, obese, and also in persons experiencing difficulty in the activities of daily living which were independent of other factors. IGF-I levels did not correlate with the subjective report of recent decline in functional status. Further studies of nocturnal GH secretion indicated that low IGF-I level was secondary to impaired growth hormone secretion. Low IGF-I levels in subjects with post-polio syndrome were corrected by physiologic doses of hGH and the response was dose dependent. Lesser dosage was required to normalize IGF-I level compared to standard dosages previously used in younger persons. Inasmuch as low IGF-I level resulting from GH deficiency is known to be associated with weakness, muscle atrophy, and decrease in aerobic work capacity, it is postulated that low IGF-I levels that occur in aging polio survivors may have an adverse effect on their neuromuscular function, and that hGH replacement may improve their functional status. Moreover, the new dose-response data may decrease the frequency of side effects in future hGH treatment of older adults.

Human growth hormone effect on serum IGF-I and muscle function in poliomyelitis survivors.

Previous work showed low insulin-like growth factor I (IGF-I) in polio survivors compared with age-matched controls and it was hypothesized that the low IGF-I was caused by the lack of growth hormone (GH) secretion. The present study asked: Is the nocturnal release of GH subnormal in polio survivors? Can the low IGF-I level be raised to the range of healthy young men (240 to 460 ng/mL) by human growth hormone (hGH) treatment? If so, what dose of hGH is required? Does the hormone treatment affect muscle function? Eleven polio survivors with evidence of postpoliomyelitis syndrome, aged 50 to 65 years, and low IGF-I levels (average IGF-I value of 170 ng/mL) were studied. The serum level of GH was measured in the first 4 hours of sleep. The serum IGF-I level was determined before and during hGH treatment at 0.0075, 0.015 or 0.03 mg/kg of ideal body weight (IBW), three times a week for successive periods of 1 month. Before and after hGH treatment, strength was determined in knee extensor and flexor muscles and the elbow flexor and elbow extensor muscles. Nocturnal GH was low in the polio survivors compared with healthy young men. Serum IGF-I was raised into the target range by either 0.0075 or 0.015 mg hGH/kg three times a week. After 3 months of hGH treatment, no consistent changes in muscle strength were observed in the study group.(ABSTRACT TRUNCATED AT 250 WORDS)

Prevalence of low plasma IGF-I in poliomyelitis survivors.

OBJECTIVE: To compare plasma levels of insulin-like growth factor-I (IGF-I, also termed somatomedin C) in polio survivors and healthy control subjects and to determine their relation to selected clinical characteristics. DESIGN: Cross sectional study. SETTING: Polio survivors living in the community recruited from the Wisconsin Polio Support Group. PARTICIPANTS: A total of 124 polio survivors (49 males and 75 females), ages 35 to 77 years, and 261 healthy control subjects (139 males and 122 females) of similar age. MEASUREMENTS: Plasma IGF-I levels were compared in polio survivors and age-matched control subjects. In the polio survivor group, the relation of IGF-I to selected clinical characteristics was examined before and after adjusting for covariates. RESULTS: Statistical analyses showed that the IGF-I concentrations were significantly lower in the polio survivors than in the controls. This difference was reflected in the means and standard errors of the two groups (0.45 +/- 0.02 vs 0.60 +/- 0.02 units/mL, P < 0.01). Plasma IGF-I below 0.35 units/mL in adults indicates little or no growth hormone secretion. In polio survivors, 38% of the plasma IGF-I values were < 0.35 units/mL compared with 19% in the healthy group. Univariate analysis showed that IGF-I in the polio survivors was significantly correlated with age, gender, and body mass index, and with dependency, pain, and difficulty in the activities of daily living (ADLs). The correlations with ADL dysfunction were independent of the correlations with age, gender, and body mass index. IGF-I level did not correlate with the subjective report of recent decline in functional status. CONCLUSION: Lower levels of IGF-I are seen in polio survivors, and this finding correlates with ADL dysfunction. The hyposomatomedinemic tendency of polio survivors may have an adverse effect on their neuromuscular function and quality of life.

Hyposomatomedinemia in men with post-poliomyelitis syndrome.

The age of onset of the post-poliomyelitis syndrome (PPS) coincides with the tendency for declining activity of the growth hormone/somatomedin C (GH/SmC) axis. The normal plasma SmC range in men before the age of 40 is 0.50 to 1.50 units/mL. After age 40 about 30% of men have a plasma SmC level below 0.35 units/mL, signifying no detectable spontaneous GH secretory pulses. Because the GH/SmC axis stimulates DNA, RNA, and protein synthesis in muscle cells and increases their size and number, a deficiency of the GH/SmC axis could theoretically contribute as a secondary factor to the occurrence or severity of the PPS. Accordingly, the authors measured the plasma SmC level in 10 men with PPS, ages 35 to 63, and in 94 healthy men of similar age. In the PPS men, 100% of the values were less than or equal to 0.40 units/mL, and 90% were less than or equal to 0.35 units/mL. The corresponding proportions in the healthy men were 40% and 27%. Analysis of variance including age as a factor showed SmC to be significantly lower in the PPS men than in the healthy men. In an additional comparison, totally immobile nursing home men did not have lowered SmC values. In fact their SmC values were slightly higher than those of healthy men of similar age. The data revealed a new biochemical feature of PPS, hyposomatomedinemia, which might play a contributory role in the pathogenesis of the syndrome.