Differentiation of tannin-containing herbal drugs by HPLC fingerprints.

A new HPLC system coupled with multiple detectors - Diode array detector (DAD), fluorescence detector (FLD), electrochemical amperometric detector (ADC) and mass spectrometry detector (MSD) was developed for the characterization and differentiation of tannin-containing herbal drugs included in The European Pharmacopoeia. The HPLC separation system consisted of an Agilent ZORBAX Eclipse XDB C18 column and a gradient water and methanol as the mobile phase which was kept at a flow rate of 0.3 mL x min(-1). Four kinds of detectors were connected by a micro-splitter valve and simultaneously recorded the response of each analytical sample. Thirty-one samples from eight kinds of tannin-containing drugs were measured using this HPLC system and their signals from all detectors were comprehensively processed via principal component analysis (PCA). The statistic result demonstrates that thirty-one batches from different herbal drugs can be reasonably identified and systematically classified by their chemical fingerprints. The proposed multi-detector HPLC method aided by chemometrics not only offers a new pattern for the study of tannin-containing herbs, but also provides a useful foundation for quality control of herbal medicines.

Specific immunotherapy has long-term preventive effect of seasonal and perennial asthma: 10-year follow-up on the PAT study.

BACKGROUND: 3-year subcutaneous specific immunotherapy (SIT) in children with seasonal allergic rhinoconjunctivitis reduced the risk of developing asthma during treatment and 2 years after discontinuation of SIT (5-year follow-up) indicating long-term preventive effect of SIT. OBJECTIVE: We evaluated the long-term clinical effect and the preventive effect of developing asthma 7-years after termination of SIT. METHODS: One hundred and forty-seven subjects, aged 16-25 years with grass and/or birch pollen allergy was investigated 10 years after initiation of a 3-year course of SIT with standardized allergen extracts of grass and/or birch or no SIT respectively. Conjunctival provocations were performed outside the season and methacholine bronchial provocations were performed during the season and winter. Asthma was assessed by clinical evaluation. RESULTS: The significant improvements in rhinoconjunctivitis and conjunctival sensitivity persisted at the 10-year follow-up. Significantly less actively treated subjects had developed asthma at 10-year follow-up as evaluated by clinical symptoms [odds ratio 2.5 (1.1-5.9)]. Patients who developed asthma among controls were 24/53 and in the SIT group 16/64. The longitudinal treatment effect when adjusted for bronchial hyper-responsiveness and asthma status at baseline including all observations at 3, 5 and 10 years follow-up (children with or without asthma at baseline, n = 189; 511 observations) was statistically significant (P = 0.0075). The odds ratio for no-asthma was 4.6 95% CI (1.5-13.7) in favor of SIT. CONCLUSION: A 3-year course of SIT with standardized allergen extracts has shown long-term clinical effects and the potential of preventing development of asthma in children with allergic rhinoconjunctivitis up to 7 years after treatment. CLINICAL IMPLICATION: Specific immunotherapy has long-term clinical effects and the potential of preventing development of asthma in children with allergic rhino conjunctivitis up to 7 years after treatment termination.

Five-year follow-up on the PAT study: specific immunotherapy and long-term prevention of asthma in children.

BACKGROUND: A 3-year course of specific immunotherapy (SIT) in children with hay fever to grass and/or birch pollen significantly reduced the risk of developing asthma. To investigate the long-term preventive effect, we performed a follow up--2 years after termination of immunotherapy. METHODS: A total of 183 children, aged 6-14 years with grass and/or birch pollen allergy could be investigated 2 years after discontinuation of SIT or no treatment. Conjunctival provocation tests (CPTs) and methacholine bronchial provocation tests were carried out during the season and winter after 5 years. The development of asthma was assessed by clinical evaluation. RESULTS: The significant improvement in hay fever and CPT results observed after 3 years of SIT persisted at the 5-year follow-up. No difference in bronchial responsiveness to methacholine was found after 5 years because of spontaneous improvement during the follow-up period in the control patients. The immunotherapy-treated children had significantly less asthma after 5 years as evaluated by clinical symptoms [odds ratio 2.68 (1.3-5.7)] in favor of SIT for prevention of development of asthma and significantly less patients reported an increase in asthma scores (P < 0.01). CONCLUSION: Immunotherapy for 3 years with standardized allergen extracts of grass and/or birch shows long-term clinical effect and preventive effect on development of asthma in children with seasonal rhinoconjunctivitis.

Effect of prepartum zeolite A supplementation on renal calcium excretion in dairy cows around calving and evaluation of a field test kit for monitoring it.

In a prospective, longitudinal study ten pregnant dry cows were randomly assigned as either experimentals or controls. The experimental period was from 28 days before expected calving until calving. Experimental cows were allocated 800 g of zeolite A daily during week 2 and 3 before expected calving. The zeolite A was given continuously via a feed dispenser. The daily calcium intake was calculated to be 60 g. Daily urine samples were obtained before, during and after the period of zeolite A supplementation. Urine pH was measured immediately whereas the remaining of samples was stored at C until analysis for calcium and creatinine. Urinary calcium/creatinine ratio was used as a measure of renal calcium excretion. Zeolite A supplementation did not affect urine pH. Renal calcium excretion peaked at the first day of supplementation, whereupon it decreased steadily to a level comparable to pre-exposure level. This could be explained by a sudden draw of calcium from the readily mobilizable pool in the bone. A commercial available water hardness test kit was evaluated as a possible cow-side test for monitoring renal calcium excretion. The repeatability of the analysis was acceptable (CV = 9.3). The accuracy of the test was good. When compared to laboratory derived urinary calcium/creatinine ratios of the same samples, a distinct linear relationship could be shown (Y = 7.9E-7 + 1.9E-8). It was therefore concluded that the water hardness test was able to monitor renal calcium excretion with an acceptable accuracy.

[Ciprofloxacin and pivampicillin in acute epididymitis in men above the age of 40 years]. / Ciprofloxacin og pivampicillin ved epididymitis acuta hos maend over 40 år.

Ciprofloxacin 500 mg orally twice daily was compared to pivampicillin 700 mg orally twice daily for 10 days in 172 men (41-85 years) with acute epididymitis. Failure of treatment occurred in 48 patients, in 15/76 (19.7%) receiving ciprofloxacin versus in 33/82 (40.2%) receiving pivampicillin (p = 0.006). Ciprofloxacin 500 mg orally twice daily is more effective than pivampicillin 700 mg orally twice daily and has fewer adverse events.

A double-blind, randomized, controlled multicentre study to compare the efficacy of ciprofloxacin with pivampicillin as oral therapy for epididymitis in men over 40 years of age.

OBJECTIVE: To compare the efficacy and safety of ciprofloxacin 500 mg orally twice daily with pivampicillin 700 mg orally twice daily for 10 days in men with acute epididymitis and over 40 years of age. PATIENTS AND METHODS: The study comprised 172 men who entered a prospective, controlled, randomized, double-blind, trial of pivampicillin and ciprofloxacin. The median (range) age of the 158 patients eligible for the efficacy analysis was 58 (41-85) years; 41% had previously had a urinary tract infection and 27% had previously had epididymitis. Only one patient had a urethral catheter and 38% were sexually active. About half of the patients were admitted to hospital. RESULTS: No bacteria could be cultured from samples in 53% of the patients; Escherichia coli could be cultured from 35% and the remaining isolates were the expected urinary pathogens. None of the patients had Gonococci and only one in each group had Chlamydia. Mycoplasma hominis was detected in three patients only and M. genitalium was detected in three, while Ureaplasma was detected in 24 (15%). The treatment failed in 48 patients; in 15 of 76 (20%) receiving ciprofloxacin and in 33 of 82 (40%) receiving pivampicillin. This corresponds to a reduction in the risk of failure of 20.5% (95% confidence limits 6.6-40.2%, P=0. 006). The principal cause of failure was an unsatisfactory clinical response requiring changed antibiotic treatment in 27 patients; adverse events were responsible for failure in 14. The in vitro resistance of cultured bacteria was low in both groups, at approximately 4%. Adverse events, mainly gastro-intestinal, occurred in 17 of 83 (21%) patients starting on ciprofloxacin and in 33 of 89 (37%) receiving pivampicillin (P=0.04). CONCLUSION: For epididymitis in men over the age of 40 years ciprofloxacin 500 mg orally twice daily is more effective than pivampicillin 700 mg orally twice daily. Furthermore, ciprofloxacin has a lower incidence of adverse events.

[Aspects of peritoneal fascia closure after surgical laparoscopy]. / Aspekte des Peritoneal-Faszienverschlusses nach operativer Laparoskopie.

Following laparoscopic surgery using trocar ports of 10 mm and more Richter's hernia and incarceration of bowel are reported. To prevent this wound closure of all layers is a must. Newly developed techniques are described. Fascia closure is accompanied by serious pain. Pain management is achieved by local anesthesia.

Secretory IgA response in oral immunotherapy. Investigation in birch pollinosis.

A rationale for oral immunotherapy (OIT) might be founded on two potential mechanisms: induction of a mucosal secretory IgA response, or induction of systemic hyporesponsiveness (oral tolerance). Previous studies have shown clinically that there is a beneficial effect of OIT in birch pollinosis, in both children and adults. During OIT, birch pollen antigens in enterocoated capsules were given to 20 adults (participating in a double-blind, placebo-controlled trial) and 10 children, all suffering from birch pollinosis. Saliva and tears (only adults) samples were collected before, during, and after OIT. Each sample was assayed for both IgA antibodies against birch pollen antigens and total IgA by enzyme-linked immunosorbent assay. IgA antibody levels were also expressed in relation to total IgA concentrations, to correct for variations in secretion and flow rate between subjects and at different times. Changes in birch-specific secretory IgA antibodies in saliva and tears could not explain the beneficial effect of OIT in birch pollinosis. Further studies in this field are warranted.

Nedocromil sodium 2% eye drops for twice-daily treatment of seasonal allergic conjunctivitis: a Swedish multicentre placebo-controlled study in children allergic to birch pollen.

This was a multicentre, double-blind, randomized group comparative study in which 77 children, aged 6-16 years, received 2% nedocromil sodium eye drops and 72 received placebo, one drop into each eye twice daily. The treatment period was 4 weeks, covering the peak birch pollen season. Prior to the start of the season, patients who had attended the clinic the previous 2 years because of seasonal allergic conjunctivitis (SAC) to birch pollen, entered a one week baseline period during which symptoms were assessed, dairy cards completed, and routine sampling of blood and urine carried out. The double-blind treatment period then commenced at the onset of the birch pollen season. Patients/parents kept daily diary record cards of eye symptom severity and concomitant therapy. Conjunctivitis was mild in both treatment groups but nedocromil sodium was more effective than placebo in controlling symptoms. During the 2-3 weeks of peak pollen counts, this therapeutic effect was statistically significant for itching (P < 0.01), watering (P < 0.05) and total symptom score (P < 0.01), but was not significant for grittiness (P = 0.08) or redness (P = 0.06). Global opinions of efficacy showed no difference between treatments, due to a high placebo effect (however, the diary card data indicated a significant improvement with nedocromil sodium). We therefore conclude that nedocromil sodium 2% eye drops, administered twice daily, is an effective treatment for SAC in children.

DNA damage induced by furocoumarin hydroperoxides plus UV (360 nm).

When irradiated at 360 nm, furocoumarins with a hydroperoxide group in a side chain efficiently give rise to a type of DNA damage that can best be explained by a photo-induced generation of hydroxyl radicals from the excited photosensitizers. The observed DNA damage profiles, i.e. the ratios of single-strand breaks, sites of base loss (AP sites) and base modifications sensitive to formamidopyrimidine--DNA glycosylase (FPG protein) and endonuclease III, are similar to the DNA damage profile produced by hydroxyl radicals generated by ionizing radiation or by xanthine and xanthine oxidase in the presence of Fe(III)--EDTA. No such damage is observed with the corresponding furocoumarin alcohols or in the absence of near-UV radiation. The damage caused by the photo-excited hydroperoxides is not influenced by superoxide dismutase (SOD) or catalase or by D2O as solvent. The presence of t-butanol, however, reduces both the formation of single-strand breaks and of base modifications sensitive to FPG protein. The cytotoxicity caused by one of the hydroperoxides in L5178Y mouse lymphoma cells is found to be dependent on the near-UV irradiation and to be much higher than that of the corresponding alcohol. Therefore the new type of photo-induced damage occurs inside cells. Intercalating photosensitizers with an attached hydroperoxide group might represent a novel and versatile class of DNA damaging agents, e.g. for phototherapy.

Pharmacokinetics of astemizole in children.

Astemizole is often administered to children in the treatment of rhinoconjunctivitis and urticaria with good efficacy and few side effects. Both astemizole and its major metabolite desmethylastemizole (DMA) are clinically effective without annoying side effects such as sedation. The pharmacokinetics in adults is well known. In three different studies we have investigated the pharmacokinetical properties of the drug in children. Study I (absorption): Thirty-eight children 8-16 years old (mean 12.6 years) and weighing 25-80 kg (mean 45 kg), with rhinoconjunctivitis due to birch pollinosis, were pretreated with either astemizole 5 mg daily or placebo for two weeks. Then, all children were treated with astemizole in doses increasing every week, i.e. 5, 10, 20 and 40 mg per day. There was a good correlation between the given dose per kg body weight and the plasma concentration of astemizole plus hydroxylated metabolites, indicating that astemizole is completely absorbed. Study II (time to reach steady state): A group of 21 children 7-18 years old (mean 13.9 years), plus 2 younger children, 2 and 5 years old, with allergy against birch- or grass pollen were treated with astemizole 10 mg daily for 12 weeks. Astemizole had reached steady-state plasma levels when the first sample was taken after 1 week, DMA reached steady state within 4 weeks. Study III (elimination half-life [t1/2 beta]): In 10 of the children from study II, t1/2 beta for astemizole plus DMA could be calculated (two samples) and was 10.8 days.(ABSTRACT TRUNCATED AT 250 WORDS)

Gingival inflammatory reactions in children with rhinoconjunctivitis due to birch pollinosis.

The present investigation was designed to study the degree of gingival inflammation in children with rhinoconjunctivitis due to birch pollinosis. Thirty-four children, 8-17 yr of age, participated. As controls, healthy classmates of the same age and sex were chosen. Dental examination was performed on three occasions: pollen season I (Spring 1987), off-season (Autumn 1987) and pollen season II (Spring 1988). The controls were examined on one occasion, mixed with the allergic children at the off-season examination. The degree of gingival inflammation was studied by determining the gingival bleeding tendency by standardized probing. Absence or presence of dental plaque was recorded after sulcular probing. In order to compensate for differences in oral hygiene level between the participants, the bleeding/plaque ratio according to Abbas et al. was calculated. The comparisons of the bleeding/plaque ratios revealed statistically significantly higher mean ratios in the allergic children during pollen seasons compared with off-season and with the controls. The results indicate that during the pollen season, children with allergic rhinoconjunctivitis exhibit an enhanced degree of gingival inflammatory reaction.

Plasticity of compensatory eye movements in rotatory tests. II. The effect of voluntary, visual, imaginary, auditory and proprioceptive mechanisms.

Two groups of 10 healthy volunteers each with a mean age of 28 years (17-39) were tested in low-frequency rotatory experiments (sinusoidal harmonic acceleration, SHA), at frequencies of 0.01-0.32 Hz. The purpose was to ascertain whether voluntary enhancement and reduction of gain and phase occurred with voluntary performances. The different tests were alertness in darkness, stationary and moving targets, imaginary stationary and moving targets in darkness, proprioceptive moving targets and acoustic stationary and moving targets. Alertness tests in darkness demonstrated a gain increase (0.5-0.7) at 0.01-0.32 Hz, and a decreasing phase lead (40-2 degrees) with increasing frequency. The alertness tests served as a reference for the other tests. In tests with stationary targets, the gain reached unity and the phase was almost 0 degrees. In imaginary stationary target tests, gain and phase were significantly increased compared with alertness. Stationary acoustic targets in darkness significantly enhanced the gain. An increased phase lead was also found. In visual suppression tests (moving target), the gain was near 0. In imaginary moving target tests, the gain decreased significantly. In darkness the gain was significantly more depressed with a proprioceptive moving target than during imaginary moving target. Testing with proprioceptive + acoustic moving target in darkness, displayed an additional gain depression and negative phase at 0.08 Hz. The conclusion is that the influence of non-vestibular mechanisms substantially affects low frequency sinusoidal rotatory testing.

Seasonal variation of the conjunctival provocation test, total and specific IgE in children with birch pollen allergy.

In 20 children with birch pollinosis, conjunctival provocation tests (CPT) with a birch pollen preparation were performed and blood was drawn on five different occasions during 1 year. From November to February there was a decrease in the activity of the atopic disease as measured by CPT (p = 0.03), total serum IGE (p = 0.02) and birch pollen-specific serum IgE (not significant). Then all parameters increased until August (p less than 0.05). From August to November there was a decrease, significant for total IgE and birch-specific IgE (p less than 0.01). The correlation between CPT and the in vitro tests was poor. The conclusion is that the seasonal variations of CPT, total and specific IgE are similar but without close correlation between CPT and the antibody levels. The priming during the season is retained at least for 1 year.

Saliva composition in asthmatic patients after treatment with two dose levels of a beta 2-adrenoceptor agonist.

Changes are known to occur in the salivary composition of asthmatic patients treated with beta 2-adrenoceptor agonists. To evaluate the precise contribution of the agonist to the impaired saliva secretion, 15 asthmatic patients, 15-23 yr old, were given two dose levels of agonist, either terbutaline or salbutamol. The lower dose, 0.15-3.0 mg/day, represented the therapeutic level used by the patients. During a wash-out period of one month, the asthma was treated with budesonide, a corticosteroid spray. Then a daily dose of 32 mg of terbutaline or salbutamol was given for one month. Samples of whole saliva, stimulated by chewing, and parotid saliva, stimulated by citric acid, were collected on three occasions: (1) at the end of the low-dose agonist treatment; (2) at the end of the wash-out period; and (3) at the end of the high-dose agonist treatment. During the high dosing the secretion rate of parotid saliva decreased and the concentrations of its total protein, amylase, hexosamine and the ratio of hexosamine/total protein were lowered. The output per minute of total protein, amylase, hexosamine, peroxidase, lysozyme, secretory IgA and potassium decreased. There were only small differences in secretion rates or saliva composition between samples collected at the end of the low-dose and at the end of the wash-out period. Thus, treatment with beta 2-adrenoceptor agonists impairs saliva secretion in asthmatics.

Effect of pollen immunotherapy on food hypersensitivity in children with birch pollinosis.

Of 72 children with severe rhinoconjunctivitis due to birch pollinosis, 67 (93%) reported sensitivity against nuts, apples, etc. The hayfever symptoms improved by immunotherapy but neither subcutaneous (n = 42) nor oral (n = 14) immunotherapy with birch pollen allergen preparations made the food sensitivity decrease significantly more than the placebo oral immunotherapy (n = 16).

Immunotherapy to deciduous tree pollens: specific IgE and IgG antibody patterns.

Thirty-nine children with birch pollinosis were given immunotherapy (IT) for 3 years with a purified, characterized and biologically standardized pollen preparation made from either birch or a mixture of birch, alder and hazel. Levels of IgE antibodies against birch, alder and oak increased early during IT (P less than 0.01) and then decreased below the initial level (P less than 0.01). Birch-specific IgG antibodies also increased (P less than 0.01) but not as rapidly as birch-specific IgE antibodies and the levels remained increased throughout IT. There were only weak correlations between immunochemical findings before IT or during early IT on one hand and the improvement during IT on the other hand. The ratio birch-specific IgG:IgE after 1.5 and 2.5 years of IT correlated with symptom scores the following season (P less than 0.01). In 60% of the children below 13 years of age, new IgE specificities developed during IT as demonstrated with crossed radioimmunoelectrophoresis. No such new IgE specificities were found in older children. These 'new sensitivities' did not appear to have any clinical implication.

Oral immunotherapy of children with rhinoconjunctivitis due to birch pollen allergy. A double blind study.

Thirty children with rhinoconjunctivitis due to birch pollinosis were treated in a double blind manner for 10 months with enteric-coated capsules containing either a birch pollen preparation (n = 14) with doses up to 1.4 X 10(6) biologic units per day or placebo (n = 16). Compared with the placebo group the actively treated children had less symptoms during the birch pollen season after 3 months of therapy (P = 0.035). Skin prick reactions decreased significantly more in the active group than in the placebo group after 10 months (P = 0.01). Conjunctival sensitivity was lower in the active group than in the placebo group after 3 months of treatment (P = 0.01) but not after 10 months. Compared with the placebo group the treated children more often increased their levels of IgG (P = 0.007) and pre-seasonal IgE (P = 0.001) against birch. There was a seasonal increase of IgE antibody level against birch in the placebo but not in the treatment group (P less than 0.001). None of the treated children developed asthma, compared with five of the untreated children. No general reactions occurred and few side effects were seen during the treatment period. We conclude that in children with birch pollinosis oral immunotherapy with high doses of a biologically potent preparation in enteric-coated capsules is effective, easy to perform, economic and safe.

Intestinal permeability as assessed with polyethyleneglycols in birch pollen allergic children undergoing oral immunotherapy.

Twenty-four children with rhinoconjunctivitis due to birch pollinosis were treated in a double-blind manner with enteric-coated capsules containing either a high dose of a birch pollen preparation (n = 11) or placebo (n = 13). The permeability of the small intestine was analysed at three different occasions with a mixture of differently sized polyethyleneglycols (PEG 400 and PEG 1000), before the start of oral immunotherapy (OIT), at the moment of maximum allergen dose, and after 3 months of therapy which was at the beginning of the pollen season. The actively treated children did not significantly change their permeability characteristics as determined from PEG recovery in the urine. By contrast, in the control group of patients the recovery of larger PEG molecules was decreased after 3 months of therapy, possibly due to the commencing pollen season. In addition, small bowel biopsies were taken at the time of maximum allergen dose from two children openly treated with OIT. Both specimens were normal.

Clinical and immunological effects of oral immunotherapy with a standardized birch pollen extract.

Oral immunotherapy (IT) was evaluated in a pilot study in two centres in children aged 8-15 years with allergic rhinoconjunctivitis. High doses (up to 20 X 10(6) BU monthly) of a defined freeze-dried birch pollen extract administered in enteric-coated gelatine capsules were given either daily for seven consecutive days every month or once weekly. Symptom scores, as assessed by sneezing, dripping and blockage of the nose, and redness, itching and swelling of the eyes, were significantly lower in treated patients compared to untreated, or placebo treated controls after 3 to 5 months of therapy. In all the 16 treated, but only in three of eight untreated patients, the scores were lower during the pollen season 1982 than during the pollen season preceding the treatment period, despite comparable pollen counts during the two seasons. One year after beginning treatment the reactivity in conjunctival provocation tests was decreased about 10-fold (P less than 0.001) in the patients receiving more than 2 X 10(5) BU monthly compared to about two-fold in patients receiving lower doses, or placebo. Increased levels of IgE antibodies directed against birch pollen were recorded in the serum and saliva of most patients after 3-4 months of active IT. In contrast, IgG antibody responses were poor in most of the patients. Side effects, particularly from the gastrointestinal tract, appeared in all treated children. In one of them a systemic reaction occurred during IT. The study indicates that properly performed oral IT with a potent birch allergen extract in enteric-coated capsules may be effective.