Analysis of A549 cell proteome alteration in response to recombinant influenza A virus nucleoprotein and its interaction with cellular proteins, a preliminary study.

Influenza A virus undergoes frequent changes of antigenicity and contributes to seasonal epidemics or unpredictable pandemics. Nucleoprotein, encoded by gene segment 5, is an internal protein of the virus and is conserved among strains of different host origins. In the current study, we analyzed the differentially expressed proteins in A549 cells transiently transfected with the recombinant nucleoprotein of influenza A virus by 2D gel electrophoresis. The resolved protein spots on gel were identified by MALDI-TOF/Mass spectrometry analysis. The majority of the host proteins detected to be differentially abundant in recombinant nucleoprotein-expressing cells as compared to vector-transfected cells are the proteins of metabolic pathways, glycolytic enzymes, molecular chaperones and cytoskeletal proteins. We further demonstrated the interaction of virus nucleoprotein with some of the identified host cellular proteins. In vitro binding assay carried out using the purified recombinant nucleoprotein (pET29a+NP-His) and A549 cell lysate confirmed the interaction between nucleoprotein and host proteins, such as alpha enolase 1, pyruvate kinase and ß-actin. The preliminary data of our study provides the information on virus nucleoprotein interaction with proteins involved in glycolysis. However, studies are ongoing to understand the significance of these interactions in modulating the host factors during virus replication.

Hyperglycemia-induced production of acute phase reactants in adipose tissue.

Chronic elevation of systemic levels of acute phase reactants and inflammatory cytokines found in patients with diabetes and the often-associated metabolic syndrome X (hypertriglyceridemia, low serum high density lipoprotein cholesterol, hypertension, and accelerated atherosclerosis) may be responsible for the increased incidence of cardiovascular problems in this population. Here we examine the contribution of adipose tissue to the systemic elevation of acute phase reactants associated with chronic hyperglycemia. We demonstrate that adipose tissue expresses a number of acute phase reactants at high levels, including serum amyloid A3 (SAA3), alphal-acid glycoprotein, the lipocalin 24p3 as well as plasminogen activator inhibitor-1 (PAI-1). Additionally, we show SAA3 is expressed at low levels under normal conditions but in the diabetic state is dramatically up-regulated in adipose tissue while down-regulated in liver. Furthermore, pro-inflammatory stimuli and high glucose can lead to the induction of SAA3 in adipose tissue in vivo as well as in the 3T3-L1 adipocyte cell line. Adipose tissue may therefore play a major role in the pathogenic sequelae of Type II diabetes, in particular the cardiovascular problems associated with prolonged hyperglycemia.

Filamin 2 (FLN2): A muscle-specific sarcoglycan interacting protein.

Mutations in genes encoding for the sarcoglycans, a subset of proteins within the dystrophin-glycoprotein complex, produce a limb-girdle muscular dystrophy phenotype; however, the precise role of this group of proteins in the skeletal muscle is not known. To understand the role of the sarcoglycan complex, we looked for sarcoglycan interacting proteins with the hope of finding novel members of the dystrophin-glycoprotein complex. Using the yeast two-hybrid method, we have identified a skeletal muscle-specific form of filamin, which we term filamin 2 (FLN2), as a gamma- and delta-sarcoglycan interacting protein. In addition, we demonstrate that FLN2 protein localization in limb-girdle muscular dystrophy and Duchenne muscular dystrophy patients and mice is altered when compared with unaffected individuals. Previous studies of filamin family members have determined that these proteins are involved in actin reorganization and signal transduction cascades associated with cell migration, adhesion, differentiation, force transduction, and survival. Specifically, filamin proteins have been found essential in maintaining membrane integrity during force application. The finding that FLN2 interacts with the sarcoglycans introduces new implications for the pathogenesis of muscular dystrophy.

Dystrobrevin and dystrophin: an interaction through coiled-coil motifs.

Dystrobrevin, a dystrophin-related and -associated protein, has been proposed to be important in the formation and maintenance of the neuromuscular junction. Dystrobrevin coprecipitates with both the acetylcholine receptor complex as well as the dystrophin glycoprotein complex. Although the nature of dystrobrevin's association with the dystrophin glycoprotein complex remains unclear, it is known that dystrobrevin binds directly to the syntrophins, a heterologous group of dystrophin-associated proteins. Using the yeast two-hybrid system to identify protein-protein interactions, we present evidence for the heterodimerization of dystrobrevin directly with dystrophin. The C terminus of dystrobrevin binds specifically to the C terminus of dystrophin. We further refined this site of interaction to these proteins' homologous coiled-coil motifs that flank their respective syntrophin-binding sites. We also show that the interaction between the dystrobrevin and dystrophin coiled-coil domains is specific and is not due to a nonspecific coiled-coil domain interaction. From the accumulated evidence of protein-protein interactions presented here and elsewhere, we propose a partially revised model of the organization of the dystrophin-associated glycoprotein complex.

Long-term follow-up of patients treated surgically for medically intractable epilepsy: results in 291 patients treated at Mayo Clinic Rochester between July 1972 and March 1985.

To assess the long-term outcome in patients who underwent surgical treatment of intractable epilepsy, we retrospectively reviewed the medical records of 291 consecutive Mayo patients treated between July 1972 and March 1985. We also evaluated the responses to a follow-up mailed questionnaire or telephone interview completed in 1992. Of the 291 patients, 245 (94% of the 261 patients known to be alive at the time of the survey) responded to the follow-up questioning. Information on the patients' neurologic status (including frequency of seizures, use of antiepileptic drugs, and self-reported assessment of functional capacity) and their overall satisfaction with the operative procedure and postoperative outcome were evaluated. Two hundred ninety patients survived the operation and were dismissed from the hospital. Of the 245 patients who responded to the follow-up survey, 41% and 58% had been free of seizures since surgical treatment and for 3 years preceding the follow-up survey, respectively. In addition, of the respondent cohort, 36% were successfully weaned off all antiepileptic drugs. Patients reported improvement in their daily functional capacity and quality of life after surgical treatment. For example, in comparison with the preoperative assessment, the patients' ability to obtain a driver's license was significantly increased, and seizure-related driving accidents, falls, and bodily injury significantly decreased. Furthermore, on the basis of current knowledge of the operation and its outcome, 85% reported that they would repeat the operative procedure. Thus, surgical treatment of intractable epilepsy is an effective option that is met with patient satisfaction.

Calcium regulation of parathyroid and C cell function in familial benign hypercalcemia.

The roles of parathyroid hormone (PTH) and calcitonin (CT) in the pathogenesis of familial benign hypercalcemia (FBH, or hypocalciuric hypercalcemia) are uncertain. Thus we performed studies in 26 patients with FBH, 12 patients with primary hyperparathyroidism (HPT), and 20 normal volunteers, to answer these questions: are plasma levels of intact or biologically active PTH frequently elevated in FBH? Is plasma intact PTH nonsuppressible during calcium infusion? Is there blunting of the C cell CT response to calcium infusion as occurs in primary HPT? We used three methods for measurement of PTH: a mid region-specific radioimmunoassay (iPTH, antiserum GP-1M), an extraction-concentration bioassay (bioPTH, stimulation of cAMP generation in osteoblastlike cells), and a two-site immunoradiometric assay (IRMA) for intact PTH. PTH levels were significantly elevated in primary HPT by all three methods, but mean PTH was normal in FBH and 85-92% of values overlapped the normal range. During 5 minute calcium infusions (2 mg Ca2+ per kg) iPTH values fell little, but bioPTH and intact PTH fell sharply in all three groups. Mean calcium-induced decreases of intact and bioPTH were indistinguishable from normal in FBH, but PTH levels generally remained elevated at 5 minutes in primary HPT. In FBH basal and postinfusion CT levels were normal. The data show that, in the majority of patients with FBH, PTH concentrations and bioactivity in blood are within the normal range and are suppressed rapidly to very low levels with further increases of calcium. The data suggest that the abnormality of parathyroid function in FBH differs from that in primary HPT.(ABSTRACT TRUNCATED AT 250 WORDS)

Distribution of serum calcium values in patients with familial benign hypercalcemia (hypocalciuric hypercalcemia): evidence for a discrete genetic defect.

One group has reported hypocalcemic individuals in families affected with familial benign hypercalcemia (FBH), suggesting either that FBH is merely an extreme of normality or that hypocalcemia is independently inherited in that kindred. To test these hypotheses, we examined the distributions of serum total calcium (Ca) values in 260 normal adults and 171 adult individuals in 21 FBH kindreds. We excluded from analysis the 21 adult probands, leaving 85 apparently affected persons (Ca, greater than 10.1 mg/dL or greater than 2.52 mmol/L) and 65 apparently unaffected individuals (Ca, less than or equal to 10.1 mg/dL or less than or equal to 2.52 mmol/L). Five FBH family members were hypocalcemic (less than 8.9 mg/dL or less than 2.22 mmol/L); of these, 3 had hypoproteinemia or hypoalbuminemia, 1 had surgical hypoparathyroidism, and 1 was pregnant (and thus excluded from further analysis). Histogram analysis suggested a bimodal distribution of Ca in the FBH families, and familial serum Ca levels were significantly elevated (P less than 0.001, rank sum). When only apparently unaffected family members were compared with normal individuals with serum Ca of 10.1 mg/dL or 2.52 mmol/L or less, the distributions were virtually identical. Our results indicate that hypocalcemia in members of families with FBH is of sporadic nongenetic origin. Furthermore, FBH is not an extreme of the normal distribution, but, instead, a clear disturbance with its own distribution about a supranormal mean serum calcium value.

Total parenteral nutrition in pregnancy: conception to delivery.

The delivery of safe and nutritionally successful total parenteral nutrition (TPN), including intravenous lipid emulsion, to a pregnant woman with Crohn's disease and short bowel syndrome from conception to delivery is reported. Maternal nutritional status improved during the pregnancy. Fetal development was normal, and a healthy, full-term infant was delivered. The placenta was normal. Specific intravenous macronutrient and micronutrient requirements during pregnancy are not well defined. Our experience has resulted in the generation of additional recommendations to optimize the intravenous nutritional therapy of this high-risk population.

The influence of an electrostatic precipitator and a mechanical filter on Rn decay products.

In the present work the effect of an HEPA filter and an electrostatic precipitator on the behaviour of Rn decay products has been studied in laboratory conditions. Both filters were found to decrease the equilibrium factor of progeny and increase the unattached fraction of decay products. In a clean air they also decreased the activity of unattached progeny. An electrostatic precipitator was found to produce condensation nuclei which were observable when no other particle sources were present. The ozone produced by the corona discharge of the filter probably has a great effect on the production of these submicron particles. The impurities of the air were found to grow the particles and thus their influence on the behaviour of Rn decay products became in some cases significant.

The control of radon progeny by air treatment devices.

The effect of air treatment devices on the behaviour of radon decay products has been studied in laboratory conditions. An HEPA filter and an electrostatic precipitator were used. Both of the filters were found to decrease the equilibrium factor of daughters and increase the unattached fraction of decay products. In a clean air they also decreased the activity of unattached daughters. The effect of the devices on the health risk caused by radon progeny was estimated by dosimetric calculations. The results corresponding to different models show considerable discrepancy, mainly due to different assumptions about the influence of unattached decay products on the dose.

Smoking and periodontal disease in the Finnish population aged 30 years and over.

The aim of this study, which was part of the Mini-Finland Oral Health Survey, was to evaluate the effect of smoking on periodontal condition in the Finnish adult population. A representative sample was drawn from the population aged 30 years and over. The total number of subjects was 8000, 90% of whom participated in the clinical examination. Periodontal diagnoses were made according to the modified Periodontal Treatment Need System (PTNS). Information about age, toothbrushing, and smoking habits was collected by personal interviews. When the sample was divided into two groups (with and without periodontal pockets), the overall risk ratio (RR) was 1.39 and was slightly higher for men than for women. A log-linear model was used to study simultaneous associations and interactions between smoking, periodontal disease, and some background factors. In this model, smoking had not significantly affected the periodontal condition.

Reproducibility of a clinical screening method for assessing gingival inflammation, pockets and plaque retentions.

The purpose of this study was to evaluate the reproducibility of the Periodontal Treatment Need System (PTNS), when plaque retentions were examined separately. A sample of 8000 persons was drawn to represent the Finnish population aged 30 yr and over. The first clinical examination (total sample) was made by a specially trained expanded-duty dental auxiliary (EDDA). During the second examination (2-6 months later), 20% of the total sample was reexamined by a dentist, and of those, every 6th patient was examined by the EDDA. Reproducibility of the PTNS was assessed as intra- and interexaminer agreement and was calculated using Cohen's kappa and weighted kappa. The intraexaminer weighted kappa values were 0.67 +/- 0.05 for the PTNS and 0.48 +/- 0.15 for plaque retentions. The interexaminer weighted kappa values between the PTNS registrations were 0.77 +/- 0.05 and 0.74 +/- 0.12 in plaque retentions when examinations were made within 1 day, and 0.53 +/- 0.05 and 0.37 +/- 0.07, respectively, when the interexaminer registrations were made with an interval of 2-6 months between them. The kappa values for intra- and interexaminer reliability reflect quite high reproducibility.

Oral hygiene, dental visits and age of denture for prevalence of denture stomatitis.

A representative sample of Finnish denture wearers (n = 3875) aged 30 or over were examined clinically and interviewed about their oral hygiene habits, use of dental services and possible prosthetic treatment during last visit to a dentist. Of these denture wearers, over 80% brushed their dentures at least once a day, and 16% visited a dentist at least once in 2 yr. For denture stomatitis the age-standardized risk ratio decreased with brushing frequency in women and increased in men. For frequency of dental visits the prevalence of denture stomatitis increased in both men and women when visits were made over 2-yr intervals. The prevalence of denture stomatitis was higher in the group with dentures over 1 yr old. It seems that the oral hygiene measures as they are generally carried out and regular dental visits are not effective enough methods in preventing denture stomatitis.

Dental caries in the adult population in Finland: I. Prevalence of dental caries.

The aim of this study was to describe the prevalence of untreated dental caries in the adult population aged 30 years and over in Finland according to sex, age, region of living, and socioeconomic status. The level of untreated caries per subject also was studied and population estimates were made for prevalence and level of untreated caries. The representative sample used in this study included 5028 dentate subjects. Data were collected using interviews and clinical examination. The prevalence of untreated caries was 55% in women, and the figure for men was 67%. In eastern Finland the figures for women and men differed significantly. The mean number of decayed teeth was 2.5 per person. Men had more decayed teeth per person than women did. The number decreased significantly with increasing socioeconomic status. The data presented in the study provide a reliable picture of the occurrence of untreated dental caries in Finnish adults and form the basis for further analyses.