Tumour necrosis factor-α/etanercept complexes in serum predict long-term efficacy of etanercept treatment in seronegative rheumatoid arthritis.

OBJECTIVE: To study whether serum levels of tumour necrosis factor-α (TNF-α), free or bound to etanercept, in biological-naïve adults with rheumatoid arthritis (RA) could predict the long-term efficacy of etanercept, measured as drug survival. METHOD: We identified 145 biological-naïve patients with RA starting treatment with etanercept at the Department of Rheumatology, Skåne University Hospital (1999-2008), of whom 16 had seronegative and 129 seropositive RA. TNF-α in serum was quantified using enzyme-linked immunosorbent assay in samples from the onset of treatment and at 6 week follow-up. Drug survival time was used to evaluate the long-term efficacy of etanercept. RESULTS: Levels of TNF-α were significantly increased at follow-up compared to at the start. At the 6 week follow-up, circulating TNF-α mainly comprised TNF-α in complex with etanercept. Longer drug survival time correlated with increased TNF-α at 6 week follow-up in the patients with seronegative RA, but not in the seropositive patients. CONCLUSION: We demonstrated that levels of circulating TNF-α increased in almost all individuals after initiation of treatment with etanercept and that this increase mainly comprised TNF-α in complex with etanercept. More importantly, this increase may predict drug survival in adults with seronegative, but not seropositive, RA and suggests that measuring TNF-α/etanercept complexes in serum may be relevant in patients with seronegative RA.

Cartilage and bone metabolism in rheumatoid arthritis. Differences between rapid and slow progression of disease identified by serum markers of cartilage metabolism.

Serum concentrations of specific cartilage and bone molecules reflecting tissue turnover were measured in two well-defined patient groups with early rheumatoid arthritis with distinctly different disease outcome to see if early differences in their levels are prognostic of the rate of joint destruction. Compared with a matched normal population, increased concentrations of cartilage oligomeric matrix protein (COMP) were found in all patients who developed rapid hip joint destruction. In contrast, levels of a putative marker of cartilage aggrecan synthesis, the chondroitin sulfate epitope 846, were increased only in patients with slow joint destruction. Levels of bone sialoprotein (BSP) were increased in both groups, as were levels of the C-propeptide of type II procollagen (CPII), a marker of collagen II synthesis. The increased concentrations of the 846 epitope in patients with slow joint destruction suggest increased aggrecan synthesis. The low levels of the 846 epitope in patients with rapid joint destruction, concomitant with elevated levels of CPII, suggest a selective increase in collagen synthesis. The elevated BSP levels indicate an increased bone turnover in both groups. Thus elevated serum levels of COMP may indicate an unfavorable prognosis for rapid joint destruction, whereas elevated 846 epitope indicates a more favorable prognosis.

Tachykinin involvement in parasympathetic nerve-evoked salivation of the ferret.

1. The tachykinin antagonist (D-Arg1, D-Cl2Phe5, Asn6, D-Trp7.9, Nle11)-substance P, injected intravenously, blocked salivary secretion from the ferret parotid and submandibular glands in response to subsequent i.v. injections of the tachykinins, substance P and neurokinin A. 2. The tachykinin antagonist reduced the parasympathetic nerve-evoked secretion of parotid and submandibular saliva by 15-20% and 35-40%, respectively. Atropine abolished the remaining secretory response. 3. The 'atropine-resistant' parasympathetic nerve-evoked secretion of saliva from the parotid and submandibular glands (about 5 and 30%, respectively, of that before administration of atropine) was abolished by the tachykinin antagonist. 4. The tachykinin antagonist was without effect on the protein concentration of parotid and submandibular saliva secreted in response to parasympathetic nerve stimulation. Parotid and submandibular saliva lacked amylase. 5. Atropine reduced the protein concentration of the submandibular saliva secreted in response to parasympathetic nerve stimulation by 50%; this was the protein concentration of substance P-evoked saliva. 6. The secretory response to methacholine and to stimulation of preganglionic sympathetic nerve fibres, tested in rats, was unaffected by the tachykinin antagonist, contra-indicating an unspecific action of the antagonist. 7. The results suggest that the neuronal release of tachykinins is probably important in the nerve-evoked secretory response of the parotid and submandibular glands.

Effects of repeated infusions of substance P and vasoactive intestinal peptide on the weights of salivary glands subjected to atrophying influences in rats.

1. The long-term influence of substance P (SP) and vasoactive intestinal peptide (VIP) on rat salivary gland weight was investigated after parasympathetic denervation or on feeding soft food. 2. The parotid gland lost about one-third of its weight within 4-5 days following parasympathetic post-ganglionic denervation or change in dietary regimen, from pellets to liquid diet, thought to reduce nerve reflex activity. 3. Daily i.v. infusions with SP or VIP diminished or largely prevented the fall in parotid gland weight, whereas infusions with pentagastrin, bethanechol and saline had no effect. The infusions were preceded by administration of alpha- and beta-adrenoceptor antagonists; these antagonists were also given to the control animals. 4. The effect of SP and VIP on the parotid gland weight appeared to be related to cell size rather than to cell number, as judged by measurements of RNA and DNA. 5. Observations on the two other major salivary glands underlined the fact that different gland types in the same animal behave differently. Parasympathetic preganglionic denervation (decentralization) lowered the weights of the sublingual and submandibular glands, whereas liquid diet only reduced the weight of the sublingual gland. SP and VIP did not affect the weights of the submandibular glands, but VIP prevented the slight fall in sublingual gland weight induced by liquid diet. 6. The present results suggest a trophic role in rats for SP and VIP on parotid glands and for VIP on sublingual glands. Such an influence may be exerted naturally as a result of their release from nerves containing these peptides around acini.

Calcitonin, calcitonin gene-related peptide, and gastrin-releasing peptide in familial thyroid medullary carcinoma.

We examined immunocytochemically the occurrence of the three peptides calcitonin, calcitonin gene-related peptide (CGRP), and gastrin-releasing peptide (GRP) in medullary carcinoma of the thyroid. We also sought to determine whether the plasma levels of these peptides were increased when stimulated with calcium and pentagastrin in familial medullary carcinoma of the thyroid (MCT). The tumor tissue from all 17 cases examined was found to exhibit calcitonin and CGRP immunoreactivity, and in 15 of the 17 cases the tumor tissue also contained GRP immunoreactivity. In 7 of the cases selected at random, an intravenous injection of calcium carbonate (2 mg/kg body weight) and pentagastrin (0.6 microgram/kg body weight) produced marked elevation in plasma levels of calcitonin but did not significantly alter the plasma levels of CGRP or GRP. We conclude that most MCT tumors contain CGRP and GRP immunoreactive cells but that the plasma levels of CGRP and GRP are not altered on stimulation. This finding is clearly in contrast to the markedly elevated calcitonin levels. Hence, determination of plasma calcitonin levels still seems to be the most appropriate diagnostic test for MCT.

Depletion of neuropeptides in rat parotid glands and declining atropine-resistant salivary secretion upon continuous parasympathetic nerve stimulation.

In rats the parasympathetic auriculo-temporal nerve on one side was continuously stimulated at 40 Hz for 20-80 min in the presence of adrenergic blockers (dihydroergotamine and propranolol) +/- atropine. During the first 10 min this gave rise to a flow of saliva from the parotid gland that in the atropinized rats amounted to 35% of that found in rats not treated with atropine, while the protein and amylase outputs were 75% of those in non-atropinized rats. The atropine-resistant secretion of fluid and proteins declined to 5-10% of the initial value within 40 min but did not cease completely even after 80 min. The marked reduction in secretory responses was not due to desensitization or exhaustion of the gland cells. The nerve stimulation reduced the parotid gland content of vasoactive intestinal peptide (VIP) and substance P (SP) to approximately 60 and 25% of that of contralateral glands after 20 and 60 min, respectively. The probable explanation for the decline in secretory response seems to be depletion of non-adrenergic, non-cholinergic transmitter(s). The present results suggest that neuropeptides are involved in the regulation of salivary secretion but provide no direct evidence that either VIP or SP is responsible for the atropine-resistant salivary secretion.

Depletion of large dense-cored vesicles from parasympathetic nerve terminals in rat parotid glands after prolonged stimulation of the auriculotemporal nerve.

Large dense-cored vesicles (60-100 nm in diameter) have been assessed electron-microscopically in terminal parasympathetic axons at acinar neuro-effector sites in rat parotid glands. Their numbers in control unstimulated glands have been compared with those in the contralateral glands of the same animals after prolonged nerve stimulation. Bilateral postganglionic sympathectomy had been undertaken 4-6 weeks previously to remove adrenergic axons from the glands. Stimulation of the postganglionic parasympathetic nerve to the gland--the auriculotemporal nerve--for 80 min at 40 Hz caused a significant depletion of large dense-cored vesicles from the terminal axons. This depletion corresponded in time and magnitude to the depletion of vasoactive intestinal peptide and substance P from the glands that had been found previously to occur under identical conditions. This adds support to the belief that the neuropeptides are stored in such vesicles and that these vesicles release their contents at neuro-effector sites as a result of propagated impulse formation in the axons.

Sensory stimulation (acupuncture) increases the release of vasoactive intestinal polypeptide in the saliva of xerostomia sufferers.

We have shown in earlier studies that xerostomia can be treated successfully with acupuncture. We also found that acupuncture stimulation can increase the concentration of neuropeptides in the saliva of healthy subjects. In this study, the concentration of the neuropeptide vasoactive intestinal polypeptide (VIP) was measured in the saliva of xerostomic patients in connection with acupuncture treatment (AP). Patients suffering from xerostomia caused by irradiation treatment, Sjögren's syndrome and other systemic disorders had been treated with acupuncture. Some of these patients showed an increase of their salivary flow rates after the AP was completed. Seventeen patients out of 65 were chosen due to their ability to produce enough saliva for the radio immunoassay (RIA) analyses to be conducted prior to the start of AP. VIP-like immunoreactivity (VIP-LI) was measured in the chewing stimulated saliva of these patients before and after the whole AP (24 sessions of 30 min each). The results showed that there was a significant increase of the concentration of VIP after the AP as compared to the measurements made before the start of the treatment (p<0.05). We concluded that the increase of neuropeptide VIP might be one of the mechanisms behind the positive effect of acupuncture on the salivary flow rates of the xerostomic patients.

Sensory stimulation (acupuncture) increases the release of calcitonin gene-related peptide in the saliva of xerostomia sufferers.

Over the last decade, several patients afflicted with xerostomia have been treated with acupuncture. Their salivary flow rates increased significantly and the improvement lasted during a long observation period. We also found that the release of several neuropeptides in the saliva of healthy subjects can be increased by acupuncture stimulation. The concentration of vasoactive intestinal polypeptide increased significantly in the saliva of xerostomic patients after acupuncture treatment. The release of the neuropeptide calcitonin gene-related peptide (CGRP) was investigated in the saliva of xerostomic patients in order to elucidate further the mechanisms of the effect of sensory stimulation (acupuncture) on the salivary secretion. CGRP-like immunoreactivity was measured with radioimmunoassay (RIA) before and after a double series of acupuncture treatment, in stimulated saliva of 14 patients who suffered from xerostomia. The results showed that the concentration of CGRP increased significantly (P<0.001) in the saliva of these patients after the end of acupuncture treatment as compared to base-line levels. Taking into consideration the influence of CGRP on the salivary flow, as well as its trophic effect, we concluded that the increased release of CGRP could be one of the factors that affect positively the salivary flow rates of xerostomic patients who were treated with acupuncture.

Parathyroid adenoma with t(1;5)(p22;q32) as the sole clonal chromosome abnormality.

Cytogenetic investigation of short-term cultures from a parathyroid adenoma revealed a t(1;5)(p22;q32) as the sole clonal chromosomal aberration. Karyotypic abnormalities have not previously been described in this tumor type.

Adjuvant chemotherapy in breast cancer.

One single course of i.v. cyclophosphamide (30 mg/kg) was given over a 6-day period to 559 mastectomized patients. During a follow-up period of up to 15 years, 241 recurrence and 234 deaths are registered in this group, and 294 recurrences and 283 deaths in the randomized control group of 577 patients. Analysis of the life-table curves shows an increased cure rate of about 10%.

Short perioperative versus long-term adjuvant chemotherapy.

A single course of cyclophosphamide IV 5 mg/kg daily for 6 days was given immediately after mastectomy to 507 patients (519 randomized controls). The relapse-free rates were significantly increased, and after 16 years the difference was 12%. In a parallel series the same adjuvant course was given 2-4 weeks after mastectomy to 52 patients (58 randomized controls). No effect of this delayed course was found. In a second study a short multidrug course was given to all patients immediately after mastectomy. One-half of the axillary node-positive cases were randomized to continue with IV CMF for 1 year. The preliminary observations show that the prolonged treatment improved the relapse-free rates significantly during the first few years. One year after mastectomy the difference was 10%; after 2 years, 9%; after 3 years, 11%; and after 4 years, 9%. The side-effects of the short course were negligible, but the side-effects of prolonged treatment were considerable and increased with increasing treatment duration. More trials are needed to find the optimum duration of adjuvant chemotherapy.

Environmental and physiological factors affecting dental fluorosis.

In addition to differences in fluoride intake and possibly to calcium deficiency or malnutrition, there are several factors which may account for individual differences in the occurrence of dental fluorosis. Disorders in acid-base balance affect the renal handling of fluoride such that, in acidosis, the excretion rate is diminished and, in alkalosis, the excretion rate is enhanced. Thus, any factor that can decrease urinary pH would be expected to increase the likelihood of dental fluorosis and vice versa. Results from studies with both rats and dogs have shown that acid-base disturbances per se, especially acidosis, adversely affect the mineralization of enamel in a manner that resembles fluorosis. It has been found that the retention and tissue levels of fluoride are increased by residence at high altitude. It has also been found that, in the absence of fluoride exposure, residence at high altitude per se can have a profound disruptive effect on amelogenesis which could be confused with fluorosis. The effects of diseases which lead to increases in urinary flow rate and water intake on the likelihood of the occurrence of dental fluorosis are also discussed.

Fluoride concentrations in rat and human tooth pulps and their possible interference with phosphatase activities.

Alkaline and acid phosphatases (p-ases), fluoride (F), and calcium (Ca) were analyzed in the pulps of rat incisors and young human pre-molars. P-ase activities were about ten times higher in the rat than in the human pulps; in the latter, high p-ase activity appeared to be connected with rapid mineralization. Average total F was 3-4 ng/mg in both rat and human pulp, while Ca showed values several times higher, although greatly varying, in human than in rat pulps. Ionized F was two to three times higher in ultrafiltrates from rat and human pulps than in the blood plasma of the species; in the rat, the F content of the ultrafiltrates increased only three to four times when plasma F was experimentally elevated over 100 times. In vitro, rat pulp alk. p-ase activity was slightly inhibited only in the range above 100 mM NaF, while acid p-ase showed weak inhibition in the region from 0.15-0.25 mM and 40-60% inhibition in the range from 10-100 mM NaF. In vivo, no effect on pulp p-ases was found, even from sublethal NaF doses.

A review of quantitative methods for studies of mineral content of intra-oral caries lesions.

Modern prospective caries studies require the measurement of small changes in tooth mineral content. Quantitative measurements of changes in mineral content in a single caries lesion is desirable. Quantitative methods can be either destructive or non-destructive. The latter type permits longitudinal studies to be conducted. Various methods available for the analysis of lesion parameters are reviewed. The basic principle of each method is summarized, and its characteristics are discussed. For each method, the correlation between the measured parameter and mineral loss, the useful range of mineral loss, the discrimination threshold, and the repeatability, are presented. Where such quantities were not available in the original papers, they were calculated from literature data. A comprehensive table of specifications of all methods is given.

Neuropeptides in the saliva of healthy subjects.

Five neuropeptides: Substance P (SP), Neurokinin A (NKA), Calcitonin Gene-Related Peptide (CGRP), Neuropeptide Y (NPY) and Vasoactive Intestinal Polypeptide (VIP), were measured in the saliva of eight subjects. The saliva was collected using different stimulation techniques: whole resting saliva, whole paraffin stimulated saliva, whole citric acid stimulated saliva and parotid saliva of different secretion rates -0.25 mL/min, 0.50 mL/min and 1.00 mL/min, also stimulated by citric acid. The neuropeptides were analysed by radioimmunoassay. The results showed that the concentration of all neuropeptides decreased significantly, two- to four-fold (CGRP up to 16-fold) in whole saliva, when the salivary secretion rates increased six- to eight-fold due to stimulation. However, the amounts of all neuropeptides released over time into the whole saliva increased two- to five-fold (ten-fold for CGRP) as the volumes of saliva increased due to chewing-stimulation as compared to resting saliva or citric acid stimulated saliva. There was also more CGRP in the resting saliva than in the citric acid stimulated saliva. The concentration of CGRP in the parotid saliva decreased three- to ten-fold when the salivary flow increased, whereas the concentration of NKA increased three- to four-fold and that of NPY almost two-fold under the same conditions. The concentrations of SP and VIP did not change in the different flows of parotid saliva. The release of all neuropeptides in the parotid saliva over time showed significant increases (3-14-fold) when the secretion rates increased except CGRP, which showed no changes at all. We concluded that neuropeptides are continuously released into the saliva. Their amounts increase with stimulation, but they are diluted by the increased volume of saliva, and they are also affected by the mode of stimulation-muscular activity leads to a greater release than citric acid stimulation. As the neuropeptides play an important role in the control of salivary secretory mechanisms, their normal occurrence and release are of fundamental importance for the understanding of the function of the salivary glands.

The influence of sensory stimulation (acupuncture) on the release of neuropeptides in the saliva of healthy subjects.

In recent studies we have shown that xerostomia (dry mouth) can be treated successfully with sensory stimulation (acupuncture). The increase of saliva secretion lasted often for at least one year. Some neuropeptides have been found to influence the secretion of saliva. The aim of this study was to investigate the mechanisms behind the effect of acupuncture on salivary secretion by measuring the release of neuropeptides in saliva under the influence of sensory stimulation. VIP-like immunoreactivity (VIP-LI), NPY-LI, SP-LI, CGRP-LI and NKA-LI were analysed in the saliva of eight healthy subjects. Manual acupuncture and acupuncture with low-frequency electrical stimulation (2 Hz) were used. The saliva was collected during 20 minutes before the start of acupuncture stimulation, then during 20 minutes while the needles were in situ and then for another 20 minutes after the needles were removed. Four different saliva sampling techniques were used: whole resting saliva, whole saliva stimulated by paraffin-chewing, whole saliva stimulated by citric acid (1%), and parotid saliva, also stimulated with citric acid (1%). The results showed significant increases in the release of CGRP, NPY and VIP both during and after acupuncture stimulation, especially in connection with electro-acupuncture. SP showed only few increases, mainly in connection with electro-acupuncture, whereas NKA generally was unaffected by the acupuncture stimulation. The sensory stimulation-induced increase in the release of CGRP, NPY and VIP in the saliva could be an indication of their role in the improvement of salivary flow rates in xerostomic patients who had been treated with acupuncture.

Inhibition of ConA-induced proliferative response in human lymphocytes by podophyllotoxin and a detergent, Tween80.

Podophyllotoxin (CPH86), at concentrations of 5 ng/ml or more, inhibited [3H]-thymidine uptake of ConA-induced proliferation of human peripheral mononuclear blood cells. Tween80, the detergent included in the anti-inflammatory drug Proresid, containing podophyllum lignan derivatives, also inhibited lymphocyte proliferation in a dose-dependent manner. Combination of the two drugs did not, or did only weakly, potentiate the inhibitory effect of each drug.

Ecology, thermodynamics and H.T. Odum's conjectures.

The central rôle of energy in all life processes has led to the development of numerous hypotheses, conjectures and theories on the relationships between thermodynamics and ecological processes. In this paper we examine the theoretical and empirical support for these developments, and in particular for the widely published set of thermodynamic conjectures developed by H.T. Odum, in which the maximum power principle is put forward as a generic feature of evolution in ecosystems. Although they are widely used, we argue that many of the ecological studies that have adopted the ideas encapsulated in Odum's work have done so without being aware of some of the fundamental problems underlying this approach. We discuss alternative ways in which a general available-work concept could be constructed for use as a numeraire in an energy-centered ecological theory or paradigm. In so doing, we examine what is meant by material accessibility and energy stocks and flows with respect to traditional food web and food chain theories, and relate these to results from the evolutionary dynamics of ecosystems. We conclude that the various forms and uses of energy bound up in essential ecosystem processes present a formidable obstacle to obtaining an operational definition of a general, aggregated available-work concept, a prerequisite for the systems approach of Odum and others. We also show that the prototypical derivations of the maximum power principle, and its interpretation, are contradicted on many scales both by empirical data and models, thereby invalidating the maximum power principle as a general principle of ecological evolution. The conclusions point to the fundamental problem of trying to describe ecosystems in a framework which has a one-dimensional currency.

Separation of the proteins of rat parotid saliva by hydrophobic-interaction chromatography.

Parasympathetically-evoked rat parotid salivary proteins were resolved into 11 peaks by hydrophobic-interaction chromatography. SDS-PAGE indicated that most peaks contained a single major component and that only one major protein was not chromatographed. The findings indicate that hydrophobic-interaction chromatography can be used in the quantification of changes in individual salivary proteins under different experimental conditions, and in the purification of individual salivary proteins.