Surface moisture increases microcracking and water vapour permeance of apple fruit skin.

Surface moisture induces microcracking in the cuticle of fruit skins. Our objective was to study the effects of surface moisture on cuticular microcracking, the permeance to water vapour and russeting in developing 'Pinova' apple fruit. Surface moisture was applied by fixing to the fruit a plastic tube containing deionized water. Microcracking was quantified by fluorescence microscopy and image analysis following infiltration with acridine orange. Water vapour permeance was determined gravimetrically using skin segments (ES) mounted in diffusion cells. Cumulative water loss through the ES increased linearly with time. Throughout development, surface moisture significantly increased skin permeance. The effect was largest during early development and decreased towards maturity. Recovery time courses revealed that following moisture treatment of young fruit for 12 days, skin permeance continued to increase until about 14 days after terminating the moisture treatment. Thereafter, skin permeance decreased over the next 28 days, then approaching the control level. This behaviour indicates gradual healing of the impaired cuticular barrier. Nevertheless, permeance still remained significantly higher compared with the untreated control. Similar patterns of permeance change were observed following moisture treatments at later stages of development. The early moisture treatment beginning at 23 DAFB resulted in russeting of the exposed surfaces. There was no russet in control fruit without a tube or in control fruit with a tube mounted for 12 days without water. The data demonstrate that surface moisture increases microcracking and water vapour permeance. This may lead to the formation of a periderm and, hence, a russeted fruit surface.

Studies on water transport through the sweet cherry fruit surface: II. Conductance of the cuticle in relation to fruit development.

Water conductance of the cuticular membrane (CM) of sweet cherry (Prunus avium L. cv. Sam) fruit during stages II and III (31-78 days after full bloom, DAFB) was investigated by gravimetrically monitoring water loss through segments of the exocarp. Segments were mounted in stainless-steel diffusion cells, filled with 0.5 ml of deionized water and incubated for 8 h at 25 +/- 2 degrees C over dry silica. Conductance was calculated by dividing the amount of water transpired per unit surface area and time by the difference in water vapor concentration across the segment (23.07 g m(-3) at 25 degrees C). Fruit mass and fruit surface area increased 4.9- and 2.8-fold between 31 and 78 DAFB, respectively. However, CM mass per unit area decreased from 3.9 to 1.5 g m(-2) and percentage of total wax content remained constant at about 31%. Stomatal density decreased from 0.8 to 0.2 mm(-2) (31-78 DAFB). Total conductance of the CM on the fruit cheek (gtot.) remained constant during stage II of development (approx. 1.38 x 10(-4) m s(-1) from 31 to 37 DAFB), increased to 1.73 x 10(-4) m s(-1) during early stage III of fruit growth (43-64 DAFB) then decreased to 0.95 x 10(-4) m s(-1) at maturity (78 DAFB). Partitioning gtot. into cuticular (gcut.) and stomatal conductance (gsto.) revealed that the relative contribution of gcut. to gtot. increased linearly from 30% to 87% of gtot. between 31 and 78 DAFB. respectively. On a whole-fruit basis, g,tot. and gcut. consistently increased up to 64 DAFB, and decreased thereafter. A significant negative linear relationship was obtained between gcut. and CM thickness, but not between the permeability coefficient (p) and CM thickness. Further, p was positively related to strain rate, suggesting that strain associated with expansion of the fruit surface increased p.

Finite dose diffusion studies: III. Effects of temperature, humidity and deposit manipulation on NAA penetration through isolated tomato fruit cuticles.

Effects of temperature, humidity, rewetting and removal of deposits on penetration of NAA [2-(1-naphthyl)acetic acid] through isolated tomato (Lycopersicon esculentum Mill) fruit cuticles were studied using a finite dose diffusion system. In this system, an aqueous 5-microliter droplet (0.1 mM NAA in 20 mM citric acid buffer) is applied to the outer surface of a cuticle, which is mounted in a glass diffusion half-cell. The cell wall surface is in contact with a receiver solution (20 mM citrate). Penetration is monitored by repeated sampling of the receiver solution. Droplets appeared dry on visual inspection within 1 h of application, but significant NAA penetration continued after droplet drying. Maximum rates of NAA penetration increased exponentially as temperature was increased (from 5 degrees to 35 degrees C), the energy of activation averaging 153 (+/- 11.6)kJ mol-1. At 35 degrees C, penetration reached a plateau within 10 h of application (at 91.1 (+/- 1.0)% of dose applied) while at 5 degrees C penetration after 800 h reached only 30.2 (+/- 7.5)%. Increasing relative humidity from 20 to 80% increased maximum rates [from 1.0 (+/- 0.21) to 2.7 (+/- 0.80)% h-1] and penetration at 120 h after application [from 36.8 (+/- 2.1) to 64.3 (+/- 3.7)%]. Rewetting deposits at 120, 240 and 360 h after application resulted in increased NAA penetration. However, amounts and rates of NAA penetration progressively decreased with each subsequent rewetting. Removal of deposits by cellulose acetate stripping at various times after droplet application resulted in a rapid decrease in NAA penetration. NAA penetration following deposit removal was always less than 6.1% of the amount of NAA applied and averaged 0.5 (+/- 0.2)% when deposits were removed immediately after droplet drying.

Photoaffinity labeling and photoaffinity cross-linking of phosphofructokinase-1 from Saccharomyces cerevisiae by 8-azidoadeninenucleotides.

Phosphofructokinase-1 from Saccharomyces cerevisiae is composed of four alpha- and four beta-subunits, each of them carrying catalytic and regulatory bindings sites for MgATP. In this paper, various photoaffinity labels, such as 8-azidoadenosine 5'-triphosphate, 8-azido-1,N6-ethenoadenosine 5'-triphosphate, and 8-N3-3'(2')-O-biotinyl-8-azidoadenosine 5'-triphosphate have been used to study their interaction with the enzyme in the dark and during irradiation. All nucleotidetriphosphates function as phosphate donor forming fructose 1,6-bisphosphate from fructose 6-phosphate. However, the kinetic analysis revealed distinctly differences between them. Photolabeling causes a decrease in enzyme activity to a similar extent, and ATP acts as competitive effector to inactivation. Three bifunctional diazidodiadeninedinucleotides (8-diN3AP4A, monoepsilon-8-diN3AP4A, and diepsilon-8-diN3AP4A) were applied for studying the spatial arrangement of the nucleotide binding sites. No cross-linking of the subunits was obtained by irradiation of the enzyme with 8-diN3AP4A. Photolabeling with diepsilon-8-diN3AP4A resulted in the formation of two alpha-beta cross-links with different mobilities in the SDS-polyacrylamide gel electrophoresis, while monoepsilon-8-diN3AP4A yielded only one alpha-beta cross-link. Because an interfacial location of the catalytic sites between two subunits is less likely, we suggest that the formation of cross-linked subunits may be the result of specific interactions of the bifunctional photolabels with regulatory sites at the interface of both subunits.

Evidence for surfactant solubilization of plant epicuticular wax.

The solubilization of isolated, reconstituted tomato (Lycopersicon esculentum Mill.) fruit and broccoli (Brassica oleracaea var. botrytis L.) leaf epicuticular waxes (ECW) by nonionic octylphenoxypolyethoxy ethanol surfactant (Triton X-100) was demonstrated in a model system by TLC and fluorescence analysis using pyrene as a fluorescent probe. ECW was solubilized at or above the surfactant critical micelle concentration; solubilization increased with an increase in micelle concentration. As shown by the fluorescence quenching of pyrene, surfactant solubilization of the ECW increased rapidly for the first 12 h, then approached a plateau, increased linearly with an increase in temperature (22--32 degrees C), and decreased linearly with the log of the polyoxyethylene chain length (range 5--40 oxyethylenes). These data are discussed in relation to surfactant effects on phytotoxicity and performance of foliar spray application of agrochemicals.

Studies on water transport through the sweet cherry fruit surface: characterizing conductance of the cuticular membrane using pericarp segments.

Water conductance of the cuticular membrane (CM) of mature sweet cherry fruit (Prunus avium L. cv. Sam) was investigated by monitoring water loss from segments of the outer pericarp excised from the cheek of the fruit. Segments consisted of epidermis, hypodermis and several cell layers of the mesocarp. Segments were mounted in stainless-steel diffusion cells with the mesocarp surface in contact with water, while the outer cuticular surface was exposed to dry silica (22 +/- 1 degrees C). Conductance was calculated by dividing the amount of water transpired per unit area and time by the difference in water vapour concentration across the segment. Conductance values had a log normal distribution with a median of 1.15 x 10(-4) m s(-1) (n=357). Transpiration increased linearly with time. Conductance remained constant and was not affected by metabolic inhibitors (1 mM NaN3 or 0.1 mM carbonylcyanide m-chlorophenylhydrazone) or thickness of segments (range 0.8-2.8 mm). Storing fruit (up to 42 d, 1 degrees C) used as a source of segments had no consistent effect on conductance. Conductance of the CM increased from cheek (1.16 +/- 0.10 x 10(-4) m s(-1)) to ventral suture (1.32 +/- 0.07 x 10(-4) m s(-1)) and to stylar end (2.53 +/- 0.17 x 10(-4) m s(-1)). There was a positive relationship (r2=0.066**; n=108) between conductance and stomatal density. From this relationship the cuticular conductance of a hypothetical astomatous CM was estimated to be 0.97 +/- 0.09 x 10(-4) m s(-1). Removal of epicuticular wax by stripping with cellulose acetate or extracting epicuticular plus cuticular wax by dipping in CHCl3/methanol increased conductance 3.6- and 48.6-fold, respectively. Water fluxes increased with increasing temperature (range 10-39 degrees C) and energies of activation, calculated for the temperature range from 10 to 30 degrees C, were 64.8 +/- 5.8 and 22.2 +/- 5.0 kJ mol(-1) for flux and vapour-concentration-based conductance, respectively.

Phase I clinical and pharmacokinetic study of titanocene dichloride in adults with advanced solid tumors.

This Phase I dose-escalation clinical trial of a lyophilized formulation of titanocene dichloride (MKT4) was conducted to determine the maximum tolerated dose, the dose-limiting toxicity (DLT), and pharmacokinetics of titanium (Ti) after a single i.v. infusion of MKT4. Forty patients with refractory solid malignancies were treated with a total of 78 courses. Using a modified Fibonacci scheme, 15 mg/m2 initial doses of titanocene dichloride were increased in cohorts of three patients up to level 11 (560 mg/m2) if DLT was not observed. The maximum tolerated dose was 315 mg/m2, and nephrotoxicity was DLT. Two minor responses (bladder carcinoma and non-small cell lung cancer) were observed. The pharmacokinetics of plasma Ti were assessed in 14 treatment courses by atomic absorption spectroscopy. The ratio for the area under the curve(0-infinity) in plasma and whole blood was 1.2. The following pharmacokinetic parameters were determined for plasma, as calculated in a two-compartment model: biological half-life t1/2beta in plasma was 22.8+/-11.2 h (xh +/- pseudo-SD), peak plasma concentration cmax approximately 30 microg/ml at a dose of 420 mg/m2, distribution volume Vss= 5.34+/-2.1 L (xa +/- SD), and a total clearance CItotal = 2.58+/-1.23 ml/min (xa +/- SD). There was a linear correlation between the area under the curve(0-infinity) of Ti in plasma and the titanocene dichloride dose administered with a correlation coefficient r2 of 0.8856. Plasma protein binding of Ti was in the 70-80% range. Between 3% and 16% of the total amount of Ti administered were renally excreted during the first 36 h. The recommended dose for Phase II evaluation is 240 mg/m2 given every 3 weeks with i.v. hydration to reduce renal toxicity.

[Current status of cataract surgery. Modern methods--internal medicine risk factors and contraindications]. / Aktueller Stand der Kataraktchirurgie. Moderne Verfahren--internistische Risikofaktoren und Kontraindikationen.

Modern cataract surgery is characterized by minimal invasive techniques that have been introduced during the past decade. These include phacoemulsification, capsulorhexis, foldable intraocular lenses and small tunnel incisions. High success rates coupled with low complication rates have resulted in a change in indications--cataract surgery is no longer performed merely to prevent blindness, but also to improve vision in patients whose professional or private visual demands are compromised by the onset of lens opacification. To ensure that their cooperation with the ophthalmic surgeon results in optimal benefit to the patient, it is important for general practitioners and internists to be conversant with the risk factors and contraindications for cataract surgery.

[Psychopharmacotherapy during pregnancy and lactation. 1: Pregnancy]. / Psychopharmakotherapie während Gravidität und Laktation. Teil 1: Gravidität.

Approximately one-third of all pregnant women take psychotropic drugs at least once during pregnancy. At the same time, there are no preparations on the market that can be considered entirely appropriate for expectant mothers. The effects of psychopharmacological therapies have exclusively been discussed in the context of their risk during the first trimester. However, treatment after this phase is not absolutely without risk, and it is striking that there are grave differences between various substances. There are currently controversial discussions going on in literature as far as the teratogenicity of lithium is concerned, especially during the formation of the heart. It is suggested that the risk for congenital malformations is increased after intrauterine lithium exposure, whereas such a risk cannot be proved for most of the antidepressants and neuroleptics. Still, it should be noted that psychopharmacology is not harmless even after the organogenesis, as intrauterine exposure during the 2nd and 3rd trimester can lead to postnatal complications. For example, floppy-infant syndrome after taking benzodiazepines, and the extrapyramidal-motor effects on the newborn after neuroleptic therapy during pregnancy should be mentioned.

[Psychopharmacotherapy in pregnancy and lactation. 2: Lactation]. / Psychopharmakotherapie während Gravidität und Laktation. Teil 2: Laktation.

Whilst the incidence of psychiatric disorders decreases during pregnancy, the risk during the postpartum period increases significantly, often leading to the necessity of psychopharmacological intervention during the puerperium, and subsequently during lactation and breast-feeding. The necessity for lithium prophylaxis in manic-depressive women after childbirth has been identified, and it is recommended that weaning rather than omission of psychopharmacological treatment is preferable during the puerperium.

The low-affinity ATP binding site of the Escherichia coli SecA dimer is localized at the subunit interface.

The homodimeric SecA protein is the ATP-dependent force generator in the Escherichia coli precursor protein translocation cascade. SecA contains two essential nucleotide binding sites (NBSs), i.e., NBS1 and NBS2 that bind ATP with high and low affinity, respectively. The photoactivatable bifunctional cross-linking agent 3'-arylazido-8-azidoadenosine 5'-triphosphate (diN3ATP) was used to investigate the spatial arrangement of the nucleotide binding sites of SecA. DiN3ATP is an authentic ATP analogue as it supports SecA-dependent precursor protein translocation and translocation ATPase. UV-induced photo-cross-linking of the diN3ATP-bound SecA results in the formation of stable dimeric species of SecA. D209N SecA, a mutant unable to bind nucleotides at NBS1, was also photo-cross-linked by diN3ATP, whereas no cross-linking occurred with the NBS2 mutant R509K SecA. We concluded that the low-affinity NBS2, which is located in the carboxyl-terminal half of SecA, is the site of crosslinking and that NBS2 binds nucleotides at or near the subunit interface of the SecA dimer.

Postpartum blues: relationship between not-protein bound steroid hormones in plasma and postpartum mood changes.

The relationship between non-bound steroid hormone levels in plasma and the occurrence of postpartum mood changes was investigated in 26 newly delivered mothers throughout the first 5 days postpartum. Studies with saliva samples had reported higher concentrations of 17 beta-estradiol and progesterone on the days of symptoms in women experiencing postpartum blues. As there had been a controversy as to how far saliva concentrations reflect free hormone levels in plasma, free hormone levels of 17 beta-estradiol and progesterone were determined in plasma using ultrafiltration. No significant difference concerning free hormone levels could be found between women with and without postpartum blues.

Preliminary results of a phase I/II trial of paclitaxel in patients with relapsed or cisplatin-refractory testicular cancer.

Paclitaxel represents a novel antitumour agent with demonstrated activity in cisplatin-sensitive tumours, particularly ovarian cancer. In addition, responses to paclitaxel have been observed in patients with cisplatin-refractory ovarian cancer. The role of paclitaxel in the treatment of testicular cancer has not been explored so far. Despite the generally high cure rates in patients with metastatic testicular cancer, patients with relapsed disease not responding to platin-based salvage chemotherapy have an extremely poor prognosis. In a phase I/II trial 10 patients with relapsed, cisplatin-refractory malignant germ-cell tumours were treated with paclitaxel as 6-h infusions (8 patients) or 3-h infusions (2 patients) at doses from 135 mg/m2 to 310 mg/m2 at 3-week intervals. Three patients achieved a response to paclitaxel, but disease recurred shortly in two patients after two and four cycles of therapy, respectively. One patient has remained in marker-negative partial response for more than 5 months. The toxicity of paclitaxel was tolerable for a dose range from 135 mg/m2 to 225 mg/m2. Granulocytopenia, WHO grades 3 and 4, occurred in all patients but was of short duration (median 3 days; range: 2-7 days). Other toxicities such as mucositis (5 patients grade 1), neurotoxicity (1 patient grade 1, 2 patients grade 2), infection (1 patient grade 3) and diarrhoea (1 patient grade 2) were not dose-limiting. There were no hypersensitivity reactions, but 1 patient developed severe myalgias during therapy with paclitaxel. Six patients with documented cisplatin-refractory disease were retreated with cisplatin-based chemotherapy after paclitaxel treatment and, in 4 of these, tumour responses of 3, 4, 5 and more than 5 months duration were achieved. In order to explore the role of paclitaxel in relapsed and/or cisplatin-refractory testicular cancer a phase II study using a 3-h infusion of 225 mg/m2 paclitaxel every 3 weeks, conducted by the German Testicular Cancer Study Group, is ongoing.

The six-minute walk--an adequate exercise test for pacemaker patients?

In many pacemaker patients bicycle and treadmill ergometry are not practicable. As an alternative, we performed a 6-minute walk on a 20-m corridor in 97 pacemaker patients, who were asked to walk as far as possible determining their speed by themselves. Results were compared with those of bicycle ergometry in 42 of these patients and with treadmill exercise of a group of 92 other pacemaker patients. In the 6-minute walk, performance and maximal heart rate were slightly lower (49 +/- 18 W; 96 +/- 23 beats/min) than in bicycle (57 +/- 16 W; 110 +/- 26 beats/min) and treadmill ergometry (50 +/- 37 W; 102 +/- 35 beats/min). A good correlation was found between walking and bicycling (r = 0.74) and in subgroups of patients with different pacemaker indications. All patients preferred the walk to bicycle ergometry considering it to be more related to daily physical activity. In conclusion, a 6-minute walk is a simple and physiological exercise test for nearly all pacemaker patients with good correlation to other types of exercise. It seems to be preferable to other tests because of its better acceptance and practicability.

Differentiation of the major flagellar antigens of Pseudomonas aeruginosa by the slide coagglutination technique.

Antisera against the two major flagellar antigens of Pseudomonas aeruginosa were obtained by immunization of rabbits with isolated flagella and absorption of contaminating antisomatic antibodies. In the conventional slide agglutination test, the pure H antisera did not agglutinate the flagellated cells of the homologous strains. The addition of protein A-bearing staphylococci to H antiserum and homologous flagellated cells, the so-called slide coagglutination, results in a rapid development of flaky clumps. H coagglutination tests of reference strains, which formerly have been H typed by long-term tube agglutination and by the indirect fluorescent-antibody technique, yielded exactly the same subdivision of the strains in H type a and H type b as the more laborious and time-consuming methods. O grouping and H typing of 181 isolates from clinical specimens revealed a free combination of the somatic and flagellar antigens. 25 OH serovars were found. The simple and rapid coagglutination technique can promote the serovar determination of P. aeruginosa, particularly for the purpose of hospital infection control.

Determination of the O-serovars of Pseudomonas aeruginosa by slide coagglutination.

Determination of the somatic (O-) antigens of Pseudomonas aeruginosa by conventional slide agglutination is frequently complicated by the barely discernible, slow reaction of native cells. For diagnostic purposes a more practical procedure, a coagglutination test, has been developed in which protein A bearing Staphylococcus aureus (ATCC 12598) cells are added to the agglutination process occurring between specific anti-O serum and native Pseudomonas aeruginosa. Compared to the conventional method, slide O-coagglutination yields larger agglutinates in a shorter mean reaction time, i.e. one minute vs four minutes. Moreover, strains not reacting in the O-agglutination method or reacting only with polyvalent anti-O serum can be grouped by O-coagglutination, and cross reactions between reference strains of different O-groups do not occur. This method facilitates O-grouping of Pseudomonas aeruginosa in epidemiological investigations.