Efficacy of different carrier gases for barrier discharge plasma generation compared to chlorhexidine on the survival of Pseudomonas aeruginosa embedded in biofilm in vitro.

Because of its antimicrobial properties, nonthermal plasma could serve as an alternative to chemical antisepsis in wound treatment. Therefore, this study investigated the inactivation of biofilm-embedded Pseudomonas aeruginosa SG81 by a surface barrier-discharged (SBD) plasma for 30, 60, 150 and 300 s. In order to optimize the efficacy of the plasma, different carrier gases (argon, argon admixed with 1% oxygen, and argon with increased humidity up to approx. 80%) were tested and compared against 0.1% chlorhexidine digluconate (CHG) exposure for 600 s. The antimicrobial efficacy was determined by calculating the difference between the numbers of colony-forming units (CFU) of treated and untreated biofilms. Living bacteria were distinguished from dead by fluorescent staining and confocal laser scanning microscopy. Both SBD plasmas and CHG showed significant antimicrobial effects compared to the untreated control. However, plasma treatment led to a higher antimicrobial reduction (argon plasma 4.9 log10 CFU/cm(2), argon with admixed oxygen 3 log10 CFU/cm(2), and with increased gas humidity 2.7 log10 CFU/cm(2) after 300 s) compared to CHG. In conclusion, SBD plasma is suitable as an alternative to CHG for inactivation of Pseudomonas aeruginosa embedded in biofilm. Further development of SBD plasma sources and research on the role of carrier gases and humidity may allow their clinical application for wound management in the future.

Efficacy of chlorhexidine, polihexanide and tissue-tolerable plasma against Pseudomonas aeruginosa biofilms grown on polystyrene and silicone materials.

BACKGROUND: The formation of biofilms is crucial in the pathogenesis of many acute and subacute microbial infections, including chronic wounds and foreign-body-related infections. Topical antimicrobial therapy with chemical antiseptics or physical treatment with tissue-tolerable plasma (TTP) may be promising to control bacterial infection. METHODS: We assessed the efficacy of 0.1% chlorhexidine digluconate (CHX), 0.02 and 0.04% polihexanide (polyhexamethylene biguanide, PHMB) and of TTP against Pseudomonas aeruginosa SG81 biofilm grown in microtitre plates (polystyrene) and on silicone materials in an artificial wound fluid. RESULTS: Overall, PHMB was as effective as CHX in reducing the total amount of biofilm (gentian violet assay) and in reducing the bacterial metabolism in biofilms (XTT assay). TTP also led to a significant reduction in colony-forming units. CONCLUSION: The antimicrobial activity of PHMB in biofilms is comparable to that of CHX. TTP could become an interesting physical alternative to chemical antisepsis in the future.

Antiseptic efficacy and tolerance of tissue-tolerable plasma compared with two wound antiseptics on artificially bacterially contaminated eyes from commercially slaughtered pigs.

AIM: To compare the tissue tolerance and efficacy of two wound antiseptics with tissue-tolerable plasma (TTP) on enucleated contaminated eyes from slaughtered pigs in order to draw consequences for the use of TTP on wounds. METHOD: The corneas of extracted eyes were contaminated with Staphylococcus aureus or Pseudomonas aeruginosa. One and 10 min after application of 10% povidone (PVP)-iodine and 0.04% polyhexanide, respectively, the eyes were rinsed with inactivating solution. To test TTP, the plasma pen meandered over the eyes at a speed of 30 mm/s and a distance of 5 mm; the eyes were then rinsed with balanced salt solution. The reduction factor was calculated by the difference between the logarithm of colony-forming units in the rinse before and after antisepsis or TTP application. RESULTS: The efficacy of TTP (reduction factor 2.4-2.9) was significantly higher (p < 0.001) than that of PVP-iodine and polyhexanide (reduction factor 1.7-2.1). CONCLUSION: TTP is more effective than the tested wound antiseptics. The lack of histological damage to the eyes of slaughtered pigs would seem to make its use as a wound antiseptic a viable alternative. In contrast to antiseptics, it supplies additional energy in the form of heat, electric fields and radicals by TTP.

Folic acid treatment in males and females with fragile-(X)-syndrome.

Ten males with the fragile X (fra(X] syndrome were treated with folic acid (10 mg/day) for 4 months in a double-blind design study. To eight heterozygotes with mental impairment and fra(X), folic acid was given for 4 months (10 mg/day) in an effort to study possible beneficial effects of folic acid. Psychological and cytogenetic testing were carried out during the trial. There was no improvement in concentration, fine motor co-ordination, or comprehension in the adult male and female patients of the study. One patient showed improvement under a control medications. In the females, improvement was seen only in the youngest patient, a 5-year-old girl. Folate treatment does not seem to be effective in fra(X) adults, but may have some effect in children of both sexes with the disorder. Cytogenetic studies using peripheral lymphocytes showed that the fra(X) frequency decreased significantly (t = 0.00856; 1% level) only in cells cultured in a folic acid-free medium but not in cells cultured in a medium with added antifolate (methotrexate). This shows a "contamination effect" of folate-free culture medium after oral folic acid treatment of these patients. The decrease of fra(X) involves primarily the early-replicating X when culturing with folic acid-free medium. A synergistic suppression effect of "external folate" and BrdU is the most likely explanation of this phenomenon.

Splanchnic sympathetic nerve activity and circulating catecholamines in the hyperthermic rat.

The mechanisms responsible for the initial rise in splanchnic vascular resistance with environmental heating are controversial, and those responsible for the subsequent fall in splanchnic resistance in the severely hyperthermic animal are unknown. Thus we examined the effect of environmental heating on plasma catecholamine concentration, splanchnic sympathetic nerve activity (SNA), and select blood chemistries. In one study, 25 male Sprague-Dawley rats (270-300 g) were assigned to one of five groups on the basis of their core temperature (Tc, 37, 39, 41, 43, or 44 degrees C) at death. Heart rate (HR), mean arterial pressure (MAP), and Tc were monitored during heat stress under alpha-chloralose anesthesia (12.5 mg.ml-1.h-1). At each predetermined Tc, an aortic blood sample was drawn and analyzed for mean plasma concentration of norepinephrine (NE), epinephrine (E), Na+, K+, and lactate. From 41 to 43 degrees C, NE and E rose significantly, and the animals became hyperkalemic and lactacidemic. In a separate study, we quantitated SNA from the greater splanchnic nerve during heat exposure of artificially respired animals anesthetized with pentobarbital sodium (50 mg/kg). MAP, splanchnic SNA, and Tc were recorded. Tc was elevated from 37.0 +/- 0.12 to 41.3 +/- 0.18 degrees C in 70 min by increase of ambient temperature to 38 degrees C in an environmental chamber. Splanchnic SNA was 54 +/- 8 spikes/s at a Tc of 37 degrees C and increased significantly as Tc exceeded 39 degrees C (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Influence of exercise training on reactivity and contractility of arterial strips from hypertensive rats.

To determine whether the lowered resting blood pressure values in hypertensive rats were associated with changes in vascular reactivity and/or contractility, helical strips were obtained from different arteries from exercise-trained (running and swimming) hypertensive rats and from running normotensive rats. Each subgroup contained nontrained controls for comparison. Changes in muscle aerobic enzymes, maximum O2 consumption, and body weight indicated that a training effect had occurred. When norepinephrine was added in a dose-response manner to the testing chamber containing helical strips from either the descending aorta, femoral artery, or renal artery, there were no significant differences in reactivity (the negative log dose-response curve) attributable to the training of hypertensive or normotensive groups. However, the trained hypertensive rats exhibited a trend for lower contractility values (dyn/mm2) in some but not all of the strips tested. From these results, we concluded that the lowered resting blood pressures associated with exercise training of hypertensive rats could not be explained by changes in vascular reactivity of their arterial strips.

Heat acclimation does not alter rat mesenteric artery response to norepinephrine.

Previous studies have shown that heat acclimation raises the temperature threshold for heat-induced splanchnic vasoconstriction in the rat (W. Haddad and M. Horowitz. Thermal Balance in Health and Disease, Advances in Pharmacological Sciences. Basel: Birkhauser, 1994, p. 203-208; M. Shochina, W. Haddad, U. Meiri, and M. Horo-witz. J. Therm. Biol. 21: 289-295, 1996). We tested the hypothesis that heat acclimation alters splanchnic resistance artery sensitivity to norepinephrine (NE). Male Sprague-Dawley rats (n = 5) were acclimated to 35 degreesC ambient temperature for 5-8 wk. Control rats (n = 5) were maintained at 22-23 degreesC ambient temperature for 5-7 wk. Small mesenteric artery segments (2- to 3-mm length, 100- to 340-micrometer ID) were isolated, cannulated at both ends, and pressurized to 50 mmHg. Artery luminal diameter was measured in response to cumulative doses of NE (10(-9) to 10(-5) M) by using video microscopy. NE dose response was measured at 37 and 43 degreesC bath temperatures. There were no differences in constriction responses to NE between acclimated and control rat arteries at either 37 or 43 degreesC. We conclude that acclimation does not alter rat mesenteric artery sensitivity to NE.

Functional and morphological changes in the eccrine sweat gland with heat acclimation.

Three adult male patas monkeys (11-15 kg) were heat acclimated by continuous exposure to an ambient temperature of 33 +/- 1 degree C at 13% relative humidity for 9 mo. During the last month, they were also exposed to 45 degrees C at 10% relative humidity for 4 h/day and 5 days/wk. Before and after 3 wk of acclimation, the animals were given a heat-tolerance test in which rectal (Tre) and mean skin (Tsk) temperatures, heart rate, and sweat rate (msw) were monitored during a 90-min exposure to 45 degrees C heat with 24% relative humidity under lenperone (1.0-1.4 mg/kg im) tranquilization. Maximal in vivo msw was also determined in response to subcutaneous injections (1 and 10% solutions) of methacholine (MCh). Before and after 9 wk and 9 mo of acclimation, sweat glands were dissected from biopsy specimens of the lateral calf, cannulated, and stimulated in vitro with MCh. Morphological measurements of isolated tubules were compared with maximal secretory rates produced by MCh stimulation. Three weeks of acclimation 1) reduced Tre and Tsk and increased msw during the heat tolerance test and 2) significantly increased maximal msw in response to MCh stimulation. Acclimation also increased (P less than 0.05) sweat gland size, as measured by tubular length and tubular volume. Maximal in vitro msw produced by MCh stimulation and msw per unit length of secretory coil also increased significantly. We conclude that heat acclimation increases the size of eccrine sweat glands and that these larger glands produce more sweat. They are also more efficient because they produce more sweat per unit length of secretory coil.

Effect of cerebrospinal fluid hyperosmolality on sweating in the heat-stressed patas monkey.

Dehydration increases the osmolality of body fluids and decreases the rate of sweating during thermal stress. By localizing osmotic stimuli to central nervous system tissues, this study assessed the role of central stimulation on sweating in a heat-stressed nonhuman primate. Lenperone-tranquilized patas monkeys (Erythrocebus patas n = 5), exposed to 41 +/- 2 degrees C, were monitored for calf sweat rate, rectal and mean skin temperatures, oxygen consumption, and heart rate during infusions (255-413 microliters) of hypertonic artificial cerebrospinal fluid (ACSF) into the third cerebral ventricle. ACSF made hypertonic with NaCl to yield osmolalities of 800 and 1,000 mosmol/kgH2O significantly decreased sweat rate compared with control ACSF (285 mosmol/kgH2O), achieving maximal reductions during infusion of 37 and 53%, respectively. Rectal temperature significantly increased during the recovery period, reaching elevations of 0.69 and 0.72 degrees C, respectively, at 20 min postinfusion. In contrast, ACSF made hypertonic with sucrose (800 mosmol/kgH2O) failed to change sweat rate or rectal temperature during infusion in three animals. Thus, intracerebroventricular infusions of hypertonic ACSF mimicked dehydration-induced effects on thermoregulation. The reduction in heat loss during infusion appeared to depend on an elevation in cerebrospinal fluid [Na+] and not osmolality per se.

Effects of CSF ANG II and AVP on sweating in the heat-stressed patas monkey.

Increasing cerebrospinal fluid [Na+] reduces sweat rate (msw) in the heat-stressed patas monkey (Erythrocebus patas). This study determined the potential role of two neuropeptides, angiotensin II (ANG II) and arginine vasopressin (AVP), in mediating this response. Artificial cerebrospinal fluid, containing either ANG II or AVP, was infused into the third cerebral ventricle of lenperone-tranquilized monkeys (n = 4) exposed to 41 +/- 2 degrees C. Solutions were infused at 16.5 microliters/min for 25 min (total vol approximately 413 microliters). ANG II (1.25, 2.5, 5, and 10 ng/microliters) tended to decrease .msw. However, during infusion, only the decline at 10 min associated with the 1.25-ng/microliters dose (26%) was different (P less than 0.004) from control. This dose elevated (P less than 0.004) core rectal temperature by 1.14 degrees C at 20 min postinfusion. In contrast, AVP (0.5 and 1.5 micrograms/microliters artificial cerebrospinal fluid) had no significant effect on .msw compared with control infusions. Both doses of AVP produced a slight but significant increase in rectal temperature of 0.14 and 0.22 degrees C, respectively, at 20 min postinfusion. In conclusion, the magnitude and time course of the change in .msw with central ANG II suggest that it does not act as the sole mediator of the decline in .msw observed with elevated cerebrospinal fluid [Na+]. The minimal effects produced by third ventricular AVP exclude this route as a means by which AVP could modulate .msw during dehydration.

Hyperthermia stimulates nitric oxide formation: electron paramagnetic resonance detection of .NO-heme in blood.

Previous experiments from our laboratory have demonstrated that severe hyperthermia results in a selective loss of splanchnic vasoconstriction. Using electron paramagnetic resonance spectroscopy to scan whole blood samples collected in vivo from the portal vein and femoral artery of conscious unrestrained rats, we observed an increase in the concentration of spectroscopy-detectable species in portal venous blood of all heat-stressed animals. These spectra consisted of at least three distinct species: one with a broad feature having an effective g factor for the unpaired electron (g) of 2.06 assigned to the copper-binding acute phase protein ceruloplasmin, and two with narrower features that evolved at core temperatures > 39 degrees C representing a semiquinone radical and .NO-heme. This heat-induced signal displays the classic nitrogen triplet hyperfine structure (nitrogen hyperfine splitting constant = 17.5 gauss, centered at g = 2.012) that is consistent with a five-coordinate heme complex and is characteristic of an unpaired electron coupled to nitrogen in the ferrous .NO-heme adduct [(alpha 2+NO) beta 3+]2. The intensity of this signal increased approximately twofold as core temperature rose to > 39 degrees C, peaking 1 h post-heat exposure at greater than threefold basal concentration. This species was not seen in corresponding arterial blood samples. This is the first demonstration that whole body hyperthermia produces increased concentrations of radicals and metal binding proteins in the venous blood of the rat and suggests that severe hyperthermia stimulates an enhanced local release of .NO within the splanchnic circulation.

Voluntary exercise and its effects on young SHR and stroke-prone hypertensive rats.

To determine whether voluntary exercise would lower resting blood pressure in spontaneously hypertensive rats (SHR) and stroke-prone spontaneously hypertensive rats (SP-SHR), two separate but interrelated investigations were undertaken. The studies were initiated when the animals were 28-35 days of age and after they were assigned to either activity or sedentary cages. The activity cages were connected to transducers and recorders that allowed the monitoring and calculation of frequency, duration, and running speed. The SHR group ran 3-7 km/day intermittently for 12 wk at high speeds (48-68 m/min), which resulted in heart rates in excess of 500 beats/min. When the SHR exercised, they seldom exceeded 33 revolutions/bout (37 m) with the majority being less than 22 revolutions/bout. This type of exercise training significantly lowered, but did not normalize, resting blood pressure by approximately 20 mmHg [nontrained (NT) = 185 +/- 5; trained (T) = 163 +/- 5 mmHg] while increasing maximum O2 consumption (VO2max) (NT = 78 +/- 2.6; T = 95 +/- 2.2 ml X min-1 X kg-1) and endurance run time (NT = 62 +/- 9.0; T = 286 +/- 15.0 min), respectively. Although SP-SHR exhibited comparable patterns of voluntary activity, the effects were not similar. First, after approximately 5 wk of consuming a special Japanese rat chow and a 1% NaCl drinking solution, cerebrovascular lesions occurred and deaths ultimately resulted in both exercising and sedentary groups. Second, although there was statistical evidence for a training effect (higher VO2max, longer VO2 test run times), voluntary exercise had no advantage in either male or female runners in lowering resting blood pressures or in improving their life-spans. Whereas voluntary activity wheel exercise or moderate forced treadmill exercise will lower resting blood pressures in young SHR populations, similar generalizations cannot be made with young SP-SHR rats.

Influence of training on the blood pressure changes during lower body negative pressure in rats.

The responses of non-trained and endurance-trained rats to conditions of lower body negative pressure (LBNP) was evaluated in normotensive, borderline hypertensive, and genetic hypertensive groups, as well as in sub-groups subjected to conditions of ventilation with 100% oxygen, systematic hemorrhaging, or sino-aortic denervations. Compared to their non-trained controls, normotensive trained rats exhibited significantly greater and faster falls in arterial blood pressure. This finding suggested a change in baroreceptor sensitivity. Related, but not statistically significant trends were observed with the hypertensive groups. Borderline hypertensive rats (DOCA injections) did not demonstrate any of these differences. Measurements of blood changes during the LBNP procedure and the effects of inspiring 100% oxygen indicated that the aortic and carotid chemoreceptors were not responsible for this training effect. After baroreceptor denervation, the group differences were abolished. In addition, the training effects were generally absent when hemorrhaging was performed, a result suggesting a difference in compliance. We have concluded from these results that endurance training will be associated with greater decreases in arterial blood pressure during LBNP than will be experienced by non-trained populations. However, the responsible mechanisms are unclear and will require further investigation.

Heat stress does not sensitize rats to the toxic effects of bacterial lipopolysaccharide.

To determine whether heat stress sensitizes rats to lipopolysaccharide (LPS), four groups were examined: saline, LPS, heat stressed+saline, and heat stressed+LPS treated rats. Saline or LPS (Escherichia coli, 5 mg.kg-1 body weight, i.v.) was given after exposure to heat and at the same time of day for nonheated rats. Survival was monitored for 24 or 48 h; samples of liver and small intestine were obtained at 24 h for histological analysis. Thermal responses were similar (P > 0.05) for the heat stressed saline and LPS treated rats: mean values for maximum colon temperature were 43.0 +/- 0.1 and 42.9 +/- 0.1 degrees C, respectively. Mortality was similar for rats exposed to heat stress+saline (11%, 2/19) and heat stress+LPS (32%, 6/19). No lethality was observed in nonheated rats given saline or LPS. Tissue damage was similar in heat stress+saline and heat stress+LPS treated rats. Liver showed mild to severe degrees of coagulative necrosis while duodenum exhibited damage to the villous tips. These findings show that severe heat stress does not markedly sensitize the rat to the lethal activity of LPS.

Influence of age and sex on the strength of bone-ligament junctions in knee joints of rats.

The bone-ligament junction strength of femur-medial collateral ligament-tibia complexes in rats was measured in situ at various ages during a two-year period. Male rats had a higher junction strength than female rats, a difference that became apparent when the animals were sixty days old and in male but not female animals subsequently paralleled the changes in body weight. However, on a bodyweight basis, the junctions were stronger in female than in male rats and this sex difference was evident at fifteen days old and persisted thereafter. Regression analysis between body weight and junction strength indicated that female rats had a significantly higher slope than males, which suggested that the sex differences were due to a hormonal factor or factors. Other measurements showed that elastic stiffness, failure energy, and collagen concentration in the ligament increased, whereas the water content of the ligament decreased with age. Most of these changes could be attriubted to the aging process and not the sex of the animal. It was concluded, however, that the strength of the insertion sites of ligaments on bones are responsive to the hormonal fluctuations that occur with aging.

[Combined cryocoagulation of ciliary body and retina in neovascular glaucoma. Initial experience]. / Kombinierte Kryokoagulation von Ziliarkörper und Retina bei Neovaskularisationsglaukom. Erste Erfahrungen.

UNLABELLED: Therapeutic success in neovascular glaucoma with conservative and surgical methods is sadly inadequate. Based on the encouraging results we have obtained with transconjunctival cyclocryothermy in primary glaucoma and with peripheral cryopexy (transconjunctival cryotherapy), we applied these two methods in combination to treat neovascular glaucoma. PATIENTS: A total of 25 eyes in 23 patients were treated with the above method. The average preoperative bulbar pressure was 43 mmHg. RESULTS: The highest average reduction of pressure was obtained in weeks 2-4 after treatment (20 mmHg); 40% of the eyes were still compensated after 4 months (below 22 mmHg), 35% showed bulbar pressures between 26-28 mmHg, and 25% showed bulbar pressures about 35 mmHg. These patients had no troublesome symptoms. In all cases, a good improvement of rubeosis iridis was observed. The combined cryotherapeutic method of treating neovascular glaucoma is efficient in controlling the internal bulbar pressure.

Knee-ligament loading properties as influenced by gravity: I. Junction with bone of 3-G rodents.

After chronic 3-G centrifugation of rats, their bone-to-ligament junctions exhibited 95 +/- 12% of the control junctions' force-sustaining capacity (F). F was actually 29 +/- 5% greater for centrifuged rats than for control rats of comparable size, as experimental animals grew to smaller body mass. This suggests that gravity determines part of F's magnitude. These junctions are, therefore, hypothesized to be weaker after development in a weightless environment. The effect was less measurable for mice. F was measured in situ as load needed to separate the knee's medial collateral ligament from the tibia of 34 male rats (Sprague-Dawley, 27-320 d of age, exposed 4-65 d), 30 control rats, 22 male mice (Swiss Webster, 35-166 d, exposed 9-56 d), and 15 control mice.

[understanding and radiation hygiene assessment of solar UV exposure in Germany]. / Erfassung und strahlenhygienische Bewertung der solaren UV-Exposition in Deutschland.

In order to monitor the natural ultraviolet (UV) radiation and to assess it from the point of view of radiation hygiene, the Federal Office for Radiation Protection (BfS) and the Federal Environmental Agency (UBA) have built up a common network for UV monitoring. Since 1993, the ground-level solar UV radiation has been measured at four sites in Germany every 6 minutes, with spectroradiometers in the wavelength range between 290 and 400 nm. Because erythema (sunborn) is of special interest in terms of radiation hygiene, the erythemal weighted irradiance was used as a measure for the biological effect of UV exposure. For this purpose, the measured spectral irradiance was multiplied by the relative erythemal weighting function, and the resulting function integrated from 290 to 400 nm. In the summer months of June and July 1999, the erythemal irradiance exceeded regularly the value of 175 mW/m2 in the south of Germany. In the north, values of not less than 160 mW/m2 were measured during this period. These values correspond to an UV index of more than 7 or 6, respectively--a high exposure regarding to the classification of the German Radiation Protection Commission (SSK), for which (absolute) sun protection is recommend. Between 1995 and 1999, the four sites were characterized by a small increase of the ground-level UV exposure of approx. 10%. Nevertheless, in consideration of the manifold factors affecting the acquired data, the measuring period of 5 years is still insufficient to demonstrate an ozone-dependent effect.