Spectroscopic investigations of fungal biomarkers after exposure to heavy ion irradiation.

The main objective of the ongoing and future space exploration missions is the search for traces of extant or extinct life (biomarkers) on Mars. One of the main limiting factors on the survival of Earth-like life is the presence of harmful space radiation, that could damage or modify also biomolecules, therefore understanding the effects of radiation on terrestrial biomolecules stability and detectability is of utmost importance. Which terrestrial molecules could be preserved in a Martian radiation scenario? Here, we investigated the potential endurance of fungal biomolecules, by exposing de-hydrated colonies of the Antarctic cryptoendolithic black fungus Cryomyces antarcticus mixed with Antarctic sandstone and with two Martian regolith analogues to increasing doses (0, 250 and 1000 Gy) of accelerated ions, namely iron (Fe), argon (Ar) and helium (He) ions. We analyzed the feasibility to detect fungal compounds with Raman and Infrared spectroscopies after exposure to these space-relevant radiations.

Modified Candy-Package technique vs Cerclage technique for refixation of the lesser trochanteric fragment in pertrochanteric femoral fractures. A biomechanical comparison of 10 specimens.

OBJECTIVES: Separation of the lesser trochanteric fragment in pertrochanteric 3-part fractures leads to a significant weakening of the medial cortical wall. Because of the attachment of the Iliopsoas muscle to this structure, the lesser trochanteric fragment tends to cranial dislocation along this muscle's action direction. Refixation of these fractures using an intramedullary nail and an additional wiring osteosynthesis can be considered an operative standard. Based on an intramedullary osteosynthesis procedure, the question was raised whether a 2-point fixation method was favourable over a 1-point method regarding the pull-out resistance of the lesser trochanteric fragment against the Iliopsoas muscle's force. METHODS: Based on an intramedullary osteosynthesis (PFNA, DePuy/Synthes/SUI), two groups á five human femora were defined depending on the refixation technique of the lesser trochanteric fragment (1-point supertrochanteric "Cable" vs 2-point super/subtrochanteric fixation "Candy-Package" performed with a 1.25-mm steel cerclage). The specimens were tested using a novel traction setup, simulating the activity pattern of the Iliopsoas muscle. The target value was the resistance of the refixated lesser trochanteric fragment against a defined pull-out force produced by the Iliopsoas muscle. The main parameters considered were the peak traction force (Fmax) and the maximum summative work (WFmax) at construct failure. RESULTS: The Fmax and WFmax displayed a significant difference in favour of the Candy-Package (2-point super/subtrochanteric fixation) group (822 N vs. 476 N, 13.8 k Nmm vs 4.4 k Nmm, respectively; P = 0.01). CONCLUSIONS: The Candy-Package technique is a method that displays significantly more resistance than a single cerclage osteosynthesis regarding fragment loosening under the application of a simulated Iliopsoas muscle force in the course of a biomechanical proximal femoral 3-part fracture model.

[Depressed fractures of the base of the middle phalanges of the fingers : Evaluation of a percutaneous, intramedullary surgical technique]. / Mittelgliedbasisimpressionsfrakturen der Finger : Evaluation eines perkutanen, intramedullären Operationsverfahrens.

BACKGROUND: Depressed fractures of the base of the middle phalanges are problematic because of frequent subluxations and centrally depressed fragments. There are two minimally invasive procedures available: 1) the less known intramedullary padding technique according to Hintringer and 2) the widely used distraction fixator of Suzuki. This article describes the technique and outcome of these two procedures. METHODOLOGY: The follow-up collective included 42 patients after treatment of a depressed fracture of the base of the middle phalanx. An intramedullary padding with percutaneous Kirschner wire retention was performed 28 times (group A) and treatment with a Suzuki fixator 14 times (group B). The study examined the hand function, the radiological results and the subjective pain level. RESULTS: According to the American Society for Surgery of the Hand (ASSH) classification 81% of the patients in group A achieved a good result but in group B the same result was achieved by only 50% of the patients. The median range of movement in the proximal interphalangeal joint was 82.5° after intramedullary padding and 47.5° after Suzuki fixator. In median, the impression was reduced from 2.35 mm to 0.5 mm in group A, but only from 1.6 mm to 1.15 mm in group B. Pain was a limiting factor in 2 out of 28 patients in group A and 1 out of 14 patients in group B. CONCLUSION: The intramedullary padding technique according to Hintringer enables good treatment of depressed fractures of the base of the middle phalanx of the finger. Repositioning of dorsal subluxations can be performed and centrally impressed fragments can be reduced better than by using the Suzuki dynamic fixator. In addition, the radiological course assessments can be assessed better than with the distraction fixator.

[Treatment of flexor tendon injuries in children]. / Behandlung von Beugesehnenverletzungen bei Kindern.

Injuries to the flexor tendons in children are less common than in adults. The clinical examination and diagnostics require extensive experience. Leaving flexor tendons untreated will result in growth disorders of the affected finger. Therefore, the indications for operative exploration of any type of open injury in a child's hand should be liberally applied. As for adults the primary treatment of flexor tendon injuries as an emergency is rarely indicated. In the recent literature various tendon suture techniques and rehabilitation protocols have been differently assessed. According to the Ulm algorithm flexor tendon injuries in children are treated by a 2-strand core stitch technique followed by a continuous circular suture. Children under 6 years of age are postoperatively immobilized for 3 weeks with a fist bandage. Children older than 6 years are treated like adults with a dynamic aftercare as described by Kleinert for 5 weeks. The results are comparable with those of other aftercare protocols.

The cluster model of ultraviolet disinfection explains tailing kinetics.

AIMS: To develop a new mathematical model derived from first principles to define the kinetics of ultraviolet disinfection and to explain the phenomenon known as tailing. The theory presented interprets tailing as the result of photoprotection due to cumulative Mie scattering effects in clustered populations of micro-organisms. METHODS AND RESULTS: Mie scattering effects at ultraviolet wavelengths are used to compute a shielding constant for each micro-organism based on the average projected diameter. An intrinsic rate constant, hypothesized to be a characteristic property of the microbial genome alone, is computed. The cluster model is fitted to tailing data from 30 ultraviolet inactivation studies and results are compared with the classic two stage multihit model. CONCLUSIONS: The cluster model demonstrates a statistically significant improvement in the mean adjusted R2 values of the tested data sets (P < 0·0001). Tailing in survival curves is the direct consequence of the Gaussian distribution of cluster sizes and the intrinsic rate constant is a real and critical parameter that defines ultraviolet susceptibility. SIGNIFICANCE AND IMPACT OF THE STUDY: The ultraviolet dose-response behaviour of micro-organisms can now be explained in terms of parameters that have physical meaning and provide deep insight into the disinfection process.

Wind-Driven Saltation: An Overlooked Challenge for Life on Mars.

Numerous studies have demonstrated that the martian surface environment is hostile to life because of its rough radiation climate and the reactive chemistry of the regolith. Physical processes such as erosion and transport of mineral particles by wind-driven saltation have hitherto not been considered as a life hazard. We report a series of experiments where bacterial endospores (spores of Bacillus subtilis) were exposed to a simulated saltating martian environment. We observed that 50% of the spores that are known to be highly resistant to radiation and oxidizing chemicals were destroyed by saltation-mediated abrasion within one minute. Scanning electron micrographs show that the spores were not only damaged by abrasion but were eradicated during the saltation process. We suggest that abrasion mediated by wind-driven saltation should be included as a factor that defines the habitability of the martian surface environment. The process may efficiently protect the martian surface from forward contamination with terrestrial microbial life-forms. Abrasion mediated by wind-driven saltation should also be considered as a major challenge to indigenous martian surface life if it exists/existed.

Pure and Oxidized Copper Materials as Potential Antimicrobial Surfaces for Spaceflight Activities.

Microbial biofilms can lead to persistent infections and degrade a variety of materials, and they are notorious for their persistence and resistance to eradication. During long-duration space missions, microbial biofilms present a danger to crew health and spacecraft integrity. The use of antimicrobial surfaces provides an alternative strategy for inhibiting microbial growth and biofilm formation to conventional cleaning procedures and the use of disinfectants. Antimicrobial surfaces contain organic or inorganic compounds, such as antimicrobial peptides or copper and silver, that inhibit microbial growth. The efficacy of wetted oxidized copper layers and pure copper surfaces as antimicrobial agents was tested by applying cultures of Escherichia coli and Staphylococcus cohnii to these metallic surfaces. Stainless steel surfaces were used as non-inhibitory control surfaces. The production of reactive oxygen species and membrane damage increased rapidly within 1 h of exposure on pure copper surfaces, but the effect on cell survival was negligible even after 2 h of exposure. However, longer exposure times of up to 4 h led to a rapid decrease in cell survival, whereby the survival of cells was additionally dependent on the exposed cell density. Finally, the release of metal ions was determined to identify a possible correlation between copper ions in suspension and cell survival. These measurements indicated a steady increase of free copper ions, which were released indirectly by cells presumably through excreted complexing agents. These data indicate that the application of antimicrobial surfaces in spaceflight facilities could improve crew health and mitigate material damage caused by microbial contamination and biofilm formation. Furthermore, the results of this study indicate that cuprous oxide layers were superior to pure copper surfaces related to the antimicrobial effect and that cell density is a significant factor that influences the time dependence of antimicrobial activity. Key Words: Contact killing-E. coli-S. cohnii-Antimicrobial copper surfaces-Copper oxide layers-Human health-Planetary protection. Astrobiology 17, 1183-1191.

Contralateral Hypoplastic Venous Draining Sinuses Are Associated with Elevated Intracranial Pressure in Unilateral Cerebral Sinovenous Thrombosis.

Variations in cerebral venous development can influence the ability to regulate drainage. In cerebral sinovenous thrombosis, these variations can be associated with elevated intracranial pressure. We present a series of pediatric patients with unilateral cerebral sinovenous thrombosis and investigate whether the contralateral venous sinus size increases the risk of developing elevated intracranial pressure. Patients diagnosed with a unilateral cerebral sinovenous thrombosis were identified by querying our institutional radiology data base. The difference in the occurrence of elevated intracranial pressure in patients with cerebral sinovenous thrombosis with and without hypoplastic venous sinuses was studied. Twelve cases of unilateral cerebral sinovenous thrombosis met the inclusion criteria and had sufficient images. Six patients had hypoplastic contralateral venous sinuses. The presence of hypoplastic contralateral venous sinus in the setting of thrombosis of a dominant sinus was associated with elevation of intracranial pressure (83% versus 0%, P = .015). Patients with cerebral sinovenous thrombosis and contralateral hypoplastic venous sinuses are at higher risk of developing elevated ICP and may benefit from screening with an ophthalmologic examination.

An Unusual Outbreak of Chlamydiosis in Commercial Turkeys Involving the Nasal Glands.

An unusual outbreak of chlamydiosis was diagnosed in 15,000, 13-wk-old organically grown turkeys housed in a semiconfinement housing system. The disease was characterized by unilateral or bilateral swelling above the eye due to mild-to-severe inflammation of the nasal glands in 3%-5% of the birds. Except for a slight drop in feed and water consumption, the birds did not exhibit any respiratory signs, morbidity, and mortality. Chlamydiosis in the turkeys was confirmed by immunofluorescence, immunohistochemistry, and PCR assay of the nasal glands. Other samples such as conjunctiva, lungs, air sacs, heart, liver, spleen, and feces were negative for chlamydia by florescence antibody test in birds submitted over several weeks. Chlamydia psittaci strain B was isolated in chicken egg embryos and typed by multilocus sequence variable number of tandem repeats analysis, multilocus sequence typing, and ompA gene sequencing as a CP3-like strain. This is the first report of a naturally occurring chlamydiosis affecting the nasal glands in turkeys.

Bovine herpesvirus-1: evaluation of genetic diversity of subtypes derived from field strains of varied clinical syndromes and their relationship to vaccine strains.

Bovine herpesvirus-1 (BoHV-1) causes significant disease in cattle. Control programs in North America incorporate vaccination with modified live viral (MLV) or killed (KV) vaccine. BoHV-1 strains are isolated from diseased animals or fetuses after vaccination. There are markers for differentiating MLV from field strains using whole-genome sequencing and analysis identifying single nucleotide polymorphisms (SNPs). Using multiple primer sets and sequencing of products permits association of BoHV-1 isolates with vaccines. To determine association between vaccine virus and strains isolated from clinical cases following vaccination, we analyzed 12 BoHV-1 isolates from animals with various clinical syndromes; 9 corresponded to BoHV-1.1 respiratory group. The remaining three corresponded to BoHV-1.2b, typically found in genital tracts of cattle. Four BoHV-1 isolates were identical to a vaccine strain; three were from post-vaccination abortion episodes with typical herpetic lesions whose dams had received MLV vaccine during pregnancy, and one from a heifer given a related MLV vaccine; Sequences of two respiratory isolates perfectly matched mutations characterizing RLB106 strain, a temperature sensitive mutant used in intranasal and parenteral vaccines. The last three respiratory strains clearly appeared related to a group of MLV vaccines. Previously the MLV vaccines were grouped into four groups based on SNPs patterns. In contrast with above-mentioned isolates that closely matched SNP patterns of their respective MLV vaccine virus, these 3 strains both lacked some and possessed a number of additional mutations compared to a group of MLV vaccine viral genome. Finding BoHV-1.2b in respiratory cases indicates focus should be given BoHV-1.2b as an emerging virus or a virus not recognized nor fully characterized in BRD.

Relationship between hair fatty acid profile, reproduction, and milk performance in Holstein Friesian cows.

The aim of this study was to analyze the relationship between the fatty acid profiles of hair of first parity cows and their reproduction and milk performance. Hair samples from 46 German Holstein cows, 50 to 62 d postpartum, were analyzed to assess the relationship. All cows were fed the same diet. The lipid composition of hair was determined by gas chromatography. The fat content in the hair was 0.86%. Hair lipids contributed 75.14% SFA, 15.81% MUFA, and 9.92% PUFA. Cows were classified into a high and a low performance group according to their a) length of the interval from calving to first insemination (CFI), b) 100-d protein yield (100-d PY), and c) length of the interval from calving to conception (CC) and 100-d PY. Cows with short CFI or high 100-d PY or short CC and high 100-d PY had significantly greater proportions of de novo synthesized C12:0 (P < 0.05) compared with the corresponding low performing groups. Increased proportions of C12:0 in the hair were associated with shorter CFI (r = -0.32, P = 0.033) and greater 100-d PY (r = 0.34, P = 0.021). Furthermore, a discriminant analysis with C12:0 as a variable best predicted short CC and high 100-d PY (error rate 33.5%). Greater proportions of de novo synthesized fatty acids in the hair, in particular C12:0, are likely to reflect a sufficient energy availability that is required for fatty acid synthesis in ruminants.

Effect of radioprotective agents in sporulation medium on Bacillus subtilis spore resistance to hydrogen peroxide, wet heat and germicidal and environmentally relevant UV radiation.

AIMS: To determine the effects of cysteine, cystine, proline and thioproline as sporulation medium supplements on Bacillus subtilis spore resistance to hydrogen peroxide (H(2)O(2)), wet heat, and germicidal 254 nm and simulated environmental UV radiation. METHODS AND RESULTS: Bacillus subtilis spores were prepared in a chemically defined liquid medium, with and without supplementation of cysteine, cystine, proline or thioproline. Spores produced with thioproline, cysteine or cystine were more resistant to environmentally relevant UV radiation at 280-400 and 320-400 nm, while proline supplementation had no effect. Spores prepared with cysteine, cystine or thioproline were also more resistant to H(2)O(2) but not to wet heat or 254-nm UV radiation. The increases in spore resistance attributed to the sporulation supplements were eliminated if spores were chemically decoated. CONCLUSIONS: Supplementation of sporulation medium with cysteine, cystine or thioproline increases spore resistance to solar UV radiation reaching the Earth's surface and to H(2)O(2). These effects were eliminated if the spores were decoated, indicating that alterations in coat proteins by different sporulation conditions can affect spore resistance to some agents. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides further evidence that the composition of the sporulation medium can have significant effects on B. subtilis spore resistance to UV radiation and H(2)O(2). This knowledge provides further insight into factors influencing spore resistance and inactivation.

Short communication: Attempts to identify Clostridium botulinum toxin in milk from three experimentally intoxicated Holstein cows.

Three adult lactating Holstein cows were injected in the subcutaneous abdominal vein with 175 ng/kg of body weight of Clostridium botulinum type C toxin (451 cow median toxic doses) to determine if this botulinum toxin crosses the blood-milk barrier. Whole blood (in sodium heparin) and clotted blood serum samples were taken at 0 min, 10 min, and 3, 6, 9, and 12 h postinoculation. Milk samples were taken at 0 min and at 3, 6, 9 and 12 h postinoculation. All samples were tested for the presence of the toxin using the mouse bioassay and immunostick ELISA test. The immunostick ELISA identified the toxin in whole blood and the mouse bioassay identified the toxin in serum at all times examined in all 3 animals. Toxin was not identified by either detection method in milk samples collected from the 3 animals. From these results, it appears that Clostridium botulinum type C toxin does not cross from the blood to the milk in detectable concentrations.

A method for extracting RNA from dormant and germinating Bacillus subtilis strain 168 endospores.

RNA was extracted from dormant and germinating Bacillus subtilis 168 spores (intact spores and chemically decoated spores) by using rapid rupture followed by acid-phenol extraction. Spore germination progress was monitored by assaying colony forming ability before and after heat shock and by reading the optical density at 600 nm. The purity, yield, and composition of the extracted RNA were determined spectrophotometrically from the ratio of absorption at 260 nm to that at 280 nm; in a 2100 BioAnalyzer, giving the RNA yield/10(8) spores or cells and the distribution pattern of rRNA components. The method reported here for the extraction of RNA from dormant spores, as well as during different phases of germination and outgrowth, has proven to be fast, efficient and simple to handle. RNA of a high purity was obtained from dormant spores and during all phases of germination and growth. There was a significant increase in RNA yield during the transition from dormant spores to germination and subsequent outgrowth. Chemically decoated spores were retarded in germination and outgrowth compared with intact spores, and less RNA was extracted; however, the differences were not significant. This method for RNA isolation of dormant, germinating, and outgrowing bacterial endospores is a valuable prerequisite for gene expression studies, especially in studies on the responses of spores to hostile environmental conditions.

Chip-based electrical detection of DNA.

A variety of methods have been developed for the detection of the binding of the complementary strand of DNA to a gene chip using electrical rather than the established optical signal techniques. Chip-based DNA sensors offer sensitivity, specificity, parallelisation and miniaturisation for the detection of selected DNA sequences or mutated genes associated with human diseases. Problems associated with the established fluorescence-based optical detection technique include the high equipment costs and the need to use sophisticated numerical algorithms to interpret the data. These problems generally limit its use to research laboratories and make it hard to adapt this detection scheme for on-site or point-of-care use. An electrical readout might be a solution to these problems. A review of a number of different approaches to achieve an electrical readout for a DNA chip is presented. The review covers various methods that are based on the use of metal nanoparticles as labels and also electrochemical methods that use polymer-modified electrodes, DNA-specific redox reporters, and DNA-mediated charge transport techniques.

Neonatal-mouse infectivity of intact Cryptosporidium parvum oocysts isolated after optimized in vitro excystation.

We reexamined the finding of Neumann et al. that intact Cryptosporidium parvum oocysts obtained after in vitro excystation were infectious for neonatal CD-1 mice. We used both established excystation protocols and our own protocol that maximized excystation. Although intact oocysts isolated after any of three protocols were infectious for neonatal CD-1 mice, the infectivity of intact oocysts isolated with our optimized excystation protocol was significantly lower than the infectivity of intact oocysts isolated after established protocols or from fresh oocysts. Excystation should not be considered a valid measure of C. parvum viability, given that it is biologically implausible for oocysts to be nonviable and yet infectious.

Hepatitis associated with a Sarcocystis canis-like protozoan in a Hawaiian monk seal (Monachus schauinslandi).

A Hawaiian monk seal (Monachus schauinslandi) died in captivity at the National Marine Fisheries Service, Kewalo Basin Facility in Honolulu, Hawaii. The animal was icteric, and the liver was friable. Microscopic lesions were detected in the colon and liver. Colonic lesions included multifocal, necrohemorrhagic colitis associated with gram-negative bacilli. The liver lesions included random hepatic necrosis and cholestasis. Asexual stages of a Sarcocystis canis-like apicomplexan were detected in hepatocytes. The parasite divided by endopolygeny. Merozoites occasionally formed rosettes around a central residual body. Ultrastructurally, merozoites lacked rhoptries. This is the first report of S. canis infection in M. schauinslandi, which is an endangered pinniped in U.S. waters.

Identification of neuropeptide-like protein gene families in Caenorhabditiselegans and other species.

Neuropeptides play critical roles in synaptic signaling in all nervous systems. Unlike classical neurotransmitters, peptidergic neurotransmitters are encoded as preproproteins that are posttranslationally processed to yield bioactive neuropeptides. To identify novel peptidergic neurotransmitters, the Caenorhabditis elegans genome was searched for predicted proteins with the structural hallmarks of neuropeptide preproproteins. Thirty-two C. elegans neuropeptide-like protein (nlp) genes were identified. The nlp genes define at least 11 families of putative neuropeptides with unique motifs; similar expressed sequence tags were identified in other invertebrate species for all 11 families. Six of these families are defined by putative bioactive motifs (FAFA, GGxYamide, MRxamide, LQFamide, LxDxamide, and GGARAF); the remaining five families are related to allatostatin, myomodulin, buccalin/drosulfakinin, orcokinin, and APGWamide neuropeptides (MGL/Famide, FRPamide, MSFamide, GFxGF, and YGGWamide families, respectively). Most C. elegans nlp gene expression is in neurons. The C. elegans nlp genes and similar genes encoding putative neuropeptides in other species are likely to play diverse roles in nervous system function.

Causes of caprine abortion: diagnostic assessment of 211 cases (1991-1998).

In an 8-year period, 1991-1998, 217 accessions of caprine abortions were submitted to the California Veterinary Diagnostic Laboratory System. Of these 217 submissions, 211 were suitable for examination in this study (6 had insufficient data). Infectious agents as the cause of abortions were found in 37% of the cases: bacterial agents were identified in 30.5%, viral agents in 2%, fungal agents in 0.5%, and protozoal agents in 4% of the cases submitted. The most common causes of abortions were Chlamydia psittaci and Coxiella burnetii infection, which accounted for 23% of all goat abortions. Mineral deficiencies were observed in 4%, fetal anomalies accounted for 3%, and leukoencephalomalacia of the brain (probable oxygen deprivation) accounted for 3% of the submissions. No diagnosis was made in 112 of the 211 submissions (53%). No lesions were noted in 104 of the submissions (49%). The other 8 submissions (4%) had histologic lesions suggestive of a bacterial agent; however, no infectious agents were identified in these cases.

Hormonal response of female goats to active immunization against a recombinant human inhibin alpha-subunit, and establishment of an enzyme-linked immunosorbent assay for caprine follicle-stimulating hormone.

The effect of selective immunosuppression of endogenous inhibin in goats on FSH, LH, progesterone and estradiol-17beta profiles was studied during the breeding and nonbreeding seasons. Eighteen adult female Boer goats were immunized against the recombinant human inhibin alpha-subunit (hINH-alpha). With the exception of estradiol, which was determined by radio-immunoassay (RIA), all plasma hormone concentrations were determined by ELISA. The ELISA for FSH presented in this paper was established in the authors' laboratory, based on an existing RIA. Mean basal concentrations of FSH were not affected by immunosuppression of endogenous inhibin, nor was there a difference in the amplitude of the pre-ovulatory FSH surge. Immunization against inhibin appears to eliminate the slight secondary rise of FSH occurring 12-20 h after the major surge associated with ovulation. The LH profiles of the immunized goats were characterized by lower basal concentrations both before and after the pre-ovulatory LH surge which itself was reduced by 50% in immunized does. By contrast, concentrations of circulating estradiol were significantly elevated after inhibin-immunization. Progesterone profiles were not affected. Extending immunization into the anoestrous season by a booster injection of hINH-alpha, implicating oestrus induction with a progestagen and eCG, produced no discernible differences in FSH and LH profiles in comparison with nonimmunized control goats. The findings suggest that in goats, paracrine factors may play a more significant role in controlling follicular activity than a feedback mechanism acting via the pituitary.