Brighter CARS hypermicroscopy via "spectral surfing" of a Stokes supercontinuum.

We present a simple technique that significantly enhances the interaction of pump pulses with a supercontinuum Stokes generated by a particular nonlinear fiber for time-gated experiments such as coherent anti-Stokes Raman scattering (CARS). The enhancement is achieved through a synchronized power-tuning/time delay scheme that we call spectral surfing. In this Letter, we introduce spectral surfing and demonstrate how its application to an economical CARS hypermicroscopy scheme increases the brightness, contrast, and spectral scanning range, while potentially reducing sample light exposure.

Spatiotemporal distributions of intestinal helminths in female lesser scaup Aythya affinis during spring migration from the upper Midwest, USA.

We examined the associations between intestinal helminth infracommunity structure and infection parameters and the age, size, and year and region of collection of 130 female lesser scaup (Aythya affinis) during their 2014-2015 spring migrations through the upper Midwest, USA. We identified a total of 647,174 individual helminths from 40 taxa, including 20 trematodes, 14 cestodes, 4 nematodes and 2 acanthocephalans parasitizing lesser scaup within the study area. Lesser scaup were each infected with 2-23 helminth taxa. One digenean, Plenosoma minimum, is reported for the first time in lesser scaup and in the Midwest. Mean trematode abundance and total helminth abundance was significantly less in 2015 than 2014, and we suspect that colder weather late in 2015 impacted the intermediate host fauna and caused the observed differences. Brillouin's species diversity of helminths was greatest in the northernmost region of the study area, which coincides with the range of a non-indigenous snail that indirectly causes annual mortality events of lesser scaup. While host age and size were not determined to be influential factors of helminth infracommunity structure, non-parametric ordination and permutational analysis of co-variance revealed that year and region of collection explained differences in helminth infracommunities. Our results suggest that spatiotemporal variations play an important role in the structure of intestinal helminth infracommunities found in migrating lesser scaup hosts, and may therefore impact host ability to build endogenous reserves at certain stopover locations in the Midwest.

GAPEWORM (SYNGAMUS SPP.) PREVALENCE IN WISCONSIN GREATER PRAIRIE CHICKENS (TYMPANUCHUS CUPIDO PINNATUS).

Under Wisconsin state law, the greater prairie chicken (GRPC; Tympanuchus cupido pinnatus) has been listed as a threatened species since 1976. In 2014-15, we conducted a pilot study to determine the prevalence and intensity of gapeworms (Syngamus spp.) in female Wisconsin GRPCs collected from 2 monitored populations. We captured 62 female GRPCs using walk-in-style traps for females and night lighting for juveniles ≥45 days of age. From these individuals, we collected 15 carcasses of radio-marked birds, most of whom died due to predation events. Through dissection, we identified gapeworm in 20% of examined carcasses and report an intensity ranging between 4 and 74 worms.

Genetic diversity among sea otter isolates of Toxoplasma gondii.

Sea otters (Enhydra lutris) have been reported to become infected with Toxoplasma gondii and at times succumb to clinical disease. Here, we determined genotypes of 39 T. gondii isolates from 37 sea otters in two geographically distant locations (25 from California and 12 from Washington). Six genotypes were identified using 10 PCR-RFLP genetic markers including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico, and by DNA sequencing of loci SAG1 and GRA6 in 13 isolates. Of these 39 isolates, 13 (33%) were clonal Type II which can be further divided into two groups at the locus Apico. Two of the 39 isolates had Type II alleles at all loci except a Type I allele at locus L358. One isolate had Type II alleles at all loci except the Type I alleles at loci L358 and Apico. One isolate had Type III alleles at all loci except Type II alleles at SAG2 and Apico. Two sea otter isolates had a mixed infection. Twenty-one (54%) isolates had an unique allele at SAG1 locus. Further genotyping or DNA sequence analysis for 18 of these 21 isolates at loci SAG1 and GRA6 revealed that there were two different genotypes, including the previously identified Type X (four isolates) and a new genotype named Type A (14 isolates). The results from this study suggest that the sea otter isolates are genetically diverse.

In vitro cell injury by oxidized low density lipoprotein involves lipid hydroperoxide-induced formation of alkoxyl, lipid, and peroxyl radicals.

Mounting evidence supports current theories linking lipoprotein oxidation to atherosclerosis. We sought the cellular biochemical mechanism by which oxidized LDL inflicts cell injury. Inhibitors of candidate pathways of cell death were used to treat human fibroblast target cells exposed to oxidized LDL.. Ebselen, which degrades lipid hydroperoxides, inhibited oxidized LDL toxicity, consistent with our recent report that 7 beta-hydroperoxycholesterol (7 beta-OOH chol) is the major cytotoxin of oxidized LDL. Intracellular chelation of metal ions inhibited, while preloading cells with iron enhanced, toxicity, Inhibition of oxidized LDL and 7 beta-OOH chol toxicity by 2-keto-4-thiolmethyl butyric acid, a putative alkoxyl radical scavenger and by vitamin E, probucol and diphenylphenylenediamine, putative scavengers of peroxyl radicals was consistent with the involvement of these radicals in the lethal sequence. Cell death was thus postulated to occur due to lipid peroxidation via a sequence involving lipid hydroperoxide-induced, iron-mediated formation of alkoxyl, lipid, and peroxyl radicals. Pathways involving other reactive oxygen species, new protein synthesis, or altered cholesterol metabolism were considered less likely, since putative inhibitors failed to lessen toxicity. Understanding the mechanism of cell injury by oxidized LDL and its toxic moiety, 7 beta-OOH chol, may indicate specific interventions in the cell injury believed to accompany vascular lesion development.

Protozoal meningoencephalitis in sea otters (Enhydra lutris): a histopathological and immunohistochemical study of naturally occurring cases.

Protozoal meningoencephalitis is considered to be an important cause of mortality in the California sea otter (Enhydra lutris). Thirty nine of 344 (11.3%) California (CA) and Washington state (WA) sea otters examined from 1985 to 2004 had histopathological evidence of significant protozoal meningoencephalitis. The aetiological agents and histopathological changes associated with these protozoal infections are described. The morphology of the actively multiplicative life stages of the organisms (tachyzoites for Toxoplasma gondii and merozoites for Sarcocystis neurona) and immunohistochemical labelling were used to identify infection with S. neurona (n=22, 56.4%), T. gondii (n=5, 12.8%) or dual infection with both organisms (n=12, 30.8%). Active S. neurona was present in all dual infections, while most had only the latent form of T. gondii. In S. neurona meningoencephalitis, multifocal to diffuse gliosis was widespread in grey matter and consistently present in the molecular layer of the cerebellum. In T. gondii meningoencephalitis, discrete foci of gliosis and malacia were more widely separated, sometimes incorporated pigment-laden macrophages and mineral, and were found predominantly in the cerebral cortex. Quiescent tissue cysts of T. gondii were considered to be incidental and not a cause of clinical disease and mortality. Protozoal meningoencephalitis was diagnosed more frequently in the expanding population of WA sea otters (10 of 31, 32.3%) than in the declining CA population (29 of 313, 9.3%). Among sea otters with protozoal meningoencephalitis, those that had displayed neurological signs prior to death had active S. neurona encephalitis, supporting the conclusion that S. neurona is the most significant protozoal pathogen in the central nervous system of sea otters.

Blood glucose monitoring in symptomatic hypoglycemia.

The relationship between blood glucose levels and the onset of hypoglycemic symptoms was studied by continuous monitoring of blood glucose levels after an oral glucose load in nine adults with normal glucose tolerance, five with chemical diabetes without symptomatic hypoglycemia, and nine with chemical diabetes with symptomatic hypoglycemia. Symptoms were associated not only with a low level of blood glucose but with a rapid fall as well. These two parameters were used to calculate a "hypoglycemic index" (defined as the fall in blood glucose during a 90-minute period prior to reaching the minimum level, divided by the value of this minimum level). The hypoglycemic index was 2.3 +/- 0.6 (mean +/- S.D.) in the group of diabetic patients with symptomatic reactive hypoglycemia and 0.7 +/- 0.3 for the other groups. This index may aid in the diagnosis of patients with symptoms of hypoglycemia and equivocally low values of blood glucose.

Tandem repeats in extrachromosomal ribosomal DNA of Dictyostelium discoideum, resulting from chromosomal mutations.

Extrachromosomal ribosomal DNA in the simple eukaryote Dictyostelium discoideum is readily separated from chromosomal DNA by orthogonal field electrophoresis (OFAGE), forming a prominent band in the 110-kb region of the gel. Here we show that mutations in at least two chromosomal genes give rise to a ladder of rDNA bands increasing in size up to about 300 kb. One of these mutations, the rrcA350 allele, which is recessive to wild type and maps to the centromere-proximal region of linkage group II, has an unstable phenotype; spontaneous revertants, which no longer exhibit the rDNA ladder, have been recovered. Another mutation rrc-351, provisionally mapped to linkage group IV, is dominant to wild type. The rDNA ladder is caused by concatamerization of a 34-kb fragment in the nontranscribed central spacer region of the 88-kb linear rDNA palindrome. Restriction enzyme analysis has revealed that each concatamer is generated by crossovers between two rDNA molecules.

Stability of hemoglobin AIc levels on repetitive determination in diabetic out-patients.

Hemoglobin (Hb) AIc levels were measured biweekly for 14 weeks in 49 diabetic out-patients and 20 nondiabetic subjects. As conventional indices of diabetes control, urine tests were performed four times per day, and plasma glucose concentrations were measured 2 h post breakfast. The diabetic group ranged in age from 15-73 yr and in duration of diabetes from 6 months to 45 yr. Eleven subjects were on diet therapy alone, three were on oral hypoglycemic agents, and 35 were on insulin therapy. The nondiabetic group ranged in age from 18-65 yr. On entry to the study, the mean (+/-SD) Hb AIc and plasma glucose levels of the diabetic group (8.60 +/- 2.11% and 142.5 +/- 99.9 mg/dl) were significantly higher than in the control group (4.68 +/- 0.60% and 102.4 +/- 21.7 mg/dl; P less than 0.001) and remained so throughout the study. The mean coefficient of variation for Hb AIc did not differ significantly between the control group and either the diet therapy or insulin therapy diabetic groups. Urine test values averaged over 2-week periods for each diabetic subject showed a high degree of stability. The mean Hb AIc levels for individual diabetic subjects correlated with the mean plasma glucose levels (r = 0.544; P less than 0.001), proportion of 2% urine tests (r = -0.798; P less than 0.001). These data provide further support for Hb AIc as a measure of diabetes control and, in addition, provide the first direct evidence that a single Hb AIc determination in a 3-month period is adequate for this purpose when the subjects are on a stable therapeutic regimen.

Reorganization of corticospinal pathways following spinal cord injury.

To assess changes in the relationship between cortical motor representation areas and their target muscles following spinal cord lesions, we studied motor evoked potentials (MEPs) to transcranial magnetic stimulation in six patients with complete spinal cord injuries at low thoracic levels and eight healthy subjects. Magnetic stimulation at rest activated a larger fraction of the motoneuron pool and evoked MEPs with shorter latencies from a larger number of scalp positions in muscles immediately rostral to the level of a spinal cord injury than in corresponding muscles in controls. The MEPs associated with maximal voluntary activation were not significantly different in the two groups. These results suggest enhanced excitability of motor pathways targeting muscles rostral to the level of a spinal transection, reflecting reorganization of motor pathways either within cortical motor representation areas or at the level of the spinal cord. The data do not allow the determination of the contribution of spinal or cortical mechanisms. However, they support the notion of a limited flexible relationship between primary motor cortex and its target muscles following alterations of normal input-output patterns.

Leg paresthesias induced by magnetic brain stimulation in patients with thoracic spinal cord injury.

We studied the induction of leg paresthesias by magnetic stimulation of the brain in seven patients with thoracic T9-12 spinal cord injury and in four normal volunteers by delivering transcranial magnetic stimulation over scalp positions 1 cm apart with a Cadwell MES-10 magnetic stimulator and an 8-shaped magnetic coil at 100% stimulus intensity. We asked subjects to report sensations felt after each stimulus. In all normal subjects, magnetic stimulation evoked sensations described as tingling or a wave descending along the leg, usually accompanied by EMG responses in leg muscles. In three of the seven patients, stimulation evoked sensations of tingling, numbness, touch, or a wave descending along the leg, lasting up to 10 seconds and referred to different parts of the legs and toes. In the patients, sensations were felt more distally the closer the site of stimulation was to the midline. Patients with leg paresthesias had less motor reorganization in abdominal muscles than those without paresthesias. These findings suggest that portions of the cortical representation areas for body parts deafferented by a complete spinal cord injury can remain related to those body parts for up to several years. A central origin of these paresthesias is probable.

Long-term botulinum toxin treatment of focal hand dystonia.

We treated focal hand dystonia in 53 patients with botulinum toxin injections for up to 6 years. Eighty-one percent of the patients improved with at least one injection session. Sixty-five percent of the injections produced transient weakness. We followed 37 of the patients for at least 2 years from the start of treatment, 24 of whom discontinued treatment because of inadequate response, loss of response, inaccessibility of a treatment provider, or the expense of the toxin. Women, who had a greater extent and longer duration of benefit than men, were more likely to continue treatment. The mean interval between injection sessions was 6 months. In most patients, we injected the toxin into the same combination of muscles at each session. The dose of toxin generally fluctuated within a range of 20 units. Side effects were mild and transient and unrelated to the long-term use of botulinum toxin. Botulinum toxin injection is safe and effective for the long-term management of focal hand dystonia.

High-dose proton pump inhibitor plus amoxycillin for the treatment or retreatment of Helicobacter pylori infection.

BACKGROUND: The combination of 120 mg of omeprazole (40 mg t.d.s.) and amoxycillin has been reported to be effective for treating H. pylori infections. METHODS: Normal volunteers with H. pylori infection received high-dose omeprazole (40 mg t.d.s.) or lansoprazole (60 mg t.d.s.) plus amoxycillin 750 mg t.d.s. for 14 days. The studies were open label and not randomized as those receiving omeprazole plus amoxycillin had previously failed lower dose omeprazole (20 mg b.d.) plus amoxycillin therapy more than 6 months previously. Those receiving lansoprazole plus amoxycillin had not been previously treated. Four to 6 weeks after ending antimicrobial therapy, H. pylori status was determined by Genta stain of gastric mucosal biopsies. RESULTS: Forty-three volunteers entered the study and 41 completed it. The overall success with high-dose proton pump inhibitor plus amoxycillin was 34.9%. For the individual regimens the per-protocol results were 48% (95% CI = 28-69%) with lansoprazole and 12.5% (95% CI = 2-38%) with omeprazole. Compliance was > 95% for both regimens. Side-effects were experienced by four lansoprazole and three omeprazole subjects, and caused two omeprazole subjects to withdraw. Cure rates were similar among different races and ethnic groups, between men and women, and between smokers and non-smokers. The level of the pre-treatment urea breath test also did not predict outcome. CONCLUSION: High-dose proton pump inhibitor plus amoxycillin combinations for treatment of H. pylori infection yielded unacceptable results, as the 95% confidence intervals did not include an 80% cure rate. These combinations do not yield consistent results worldwide and cannot be recommended as primary therapy.

Metronidazole, ranitidine and clarithromycin combination for treatment of Helicobacter pylori infection (modified Bazzoli's triple therapy).

BACKGROUND: Multi-drug regimens are generally required to reliably cure Helicobacter pylori infection. Metronidazole, clarithromycin and omeprazole has proven to be an effective combination therapy with a cure rate of 90% or greater. METHODS: We evaluated a 14-day combination regimen for H.pylori infection consisting of metronidazole 500 mg b.d., clarithromycin 250 mg b.d. and ranitidine 300 mg b.d. (MRC) instead of omeprazole. Ranitidine alone was continued for an additional 4 weeks. H. pylori status was determined by rapid urease testing, histopathology using the Genta stain, and by culture at entry and 4 weeks after completing antimicrobial therapy. RESULTS: Twenty-seven patients with documented peptic ulcer disease and H. pylori infection were treated. Five had previously failed macrolide-based antimicrobial therapy; none had received metronidazole. All ulcers were healed at week 6 except one patient taking naproxen; his H. pylori infection was cured. Overall, H. pylori infection was cured in 78% (95% CI = 58-91%). In patients with clarithromycin-sensitive isolates, the cure rate was 20 of 23 (87%, 95% C.I. = 66-97%); only one of four patients (25%) with clarithromycin-resistant isolates was cured. In contrast, four of five patients with metronidazole-resistant isolates were cured (80%). In patients with isolates sensitive to both antibiotics, the cure rate was 16 of 18 (89%, 95% C.I. = 65-99%). Mild side effects were reported by 27%, including diarrhoea and altered taste. Compliance averaged 98%. CONCLUSION: These results suggest that the combination of metronidazole, ranitidine and clarithromycin results in high cure rates in patients with clarithromycin-sensitive isolates. Omeprazole may not be required for Bazzoli's triple therapy; and large multicentre comparative trials are indicated.

Metronidazole, omeprazole and clarithromycin: an effective combination therapy for Helicobacter pylori infection.

BACKGROUND: Successful treatment of Helicobacter pylori infection results in cure of peptic ulcer disease. Multi-drug regimens are needed to cure this infection. We studied the effectiveness and side effect profile of two antibiotics active against Helicobacter pylori, metronidazole and clarithromycin, combined with omeprazole. METHODS: We evaluated a combination therapy for H. pylori infection consisting of metronidazole (500 mg b.d.), omeprazole (20 mg b.d.), and clarithromycin (250 mg b.d.) for 2 weeks, followed by ranitidine 300 mg daily for 4 weeks. RESULTS: Thirty-three patients with documented H. pylori infection were studied. Twenty had previously failed antimicrobial therapy, including one with metronidazole-based triple therapy and eight with macrolide-based therapy (five with clarithromycin-based therapy), and 11 with amoxycillin, tetracycline, and bismuth. H. pylori status was determined by histopathology using the Genta stain and by culture. H. pylori status was determined at entry and 4 weeks after completing antimicrobial therapy. The H. pylori infection was cured in 88% (95% CI = 72%-96%) including 90% of those who had failed previous anti-H. pylori therapies. Mild side effects were reported by 18%. CONCLUSION: We conclude that the combination of metronidazole, omeprazole and clarithromycin is an effective treatment for H. pylori infection.

Further development and automation of a high pressure liquid chromatography method for the determination of glycosylated hemoglobins.

A previously described high pressure liquid chromatography system for the determination of glycosylated hemoglobin concentrations has been automated and simplified. With this methodology it is possible to perform up to 60 analyses per day for hemoglobin A1a+b% and hemoglobin A1c%. Where an estimate of the total fast hemoglobin alone is required, then a considerably greater number of analyses can be performed. The individual values are calculated directly with an electronic integrator. The mean coefficient of variation of the duplicate determinations of 48 samples was 0.63 +/- 0.83% (mean +/- SD). Aliquots of pooled hemolysates have been maintained in liquid nitrogen at -90 degrees C and run at the beginning and end of each daily analytical run over an 18-mo period. Both the inter- and intra-run coefficients of variation of these values have remained consistently less than 3%. Therefore, the methodology offers a reliable and accurate method of containing glycosylated hemoglobin values for clinical use.

A rapid method for the determination of glycosylated hemoglobins using high pressure liquid chromatography.

Hemoglobin (Hb) Alc is a minor component of Hb found in normal individuals but elevated two or threefold in patients with diabetes mellitus. Limited studies have suggested that the level of Hb Alc is proportional to the integrated concentration of glucose over time. Thus it could serve as an index of hyperglycemia. Its measurement may enable a more objective approach to assessing whether or not the control of hyperglycemia can be correlated with the severity of complications of diabetes. Large scale clinicab studies of Hb Alc have not been undertaken for lack of a rapid assay system. This article describes a method of high pressure liquid chromatography (HPLC) which enables the isolation of Hb Alc in 27 min using only 12 microgram of Hb (100 microliter of blood) and a second method for the isolation of total fast Hb components (also elevated in diabetes) in 11 min. Using the first method, a total of 36 assays were performed on the blood of a single normal volunteer over a one month period. the mean level of Hb Alc was 4.95 +/- 0.12% (SD) +/- 0.02% (SEM), while the coefficient of variation (C.V.) was 2.4%. The mean Hb Alc & b level was 1.65 +/- 0.06% +/- 0.01% (C.V. = 3.6%). Values for Hb Alc in 10 normal individuals were 5.06 (mean) +/- 0.32% (SD) +/- 0.01% (SEM). Hb Alc values in 15 patients with diabetes mellitus ranged from 6.8 to 20.0%. The second method was designed to assay Hb Ala, Hb Alb, and Hb Alc as a single peak and yielded results identical to the sum of these components as determined by the first method ( r = 0.98; p less than 0.001).

First record of an hypopus (Acari: Hypoderatidae) from a jaeger (Aves: Charadriiformes: Stercorariidae).

Thalassornectes (Alcidectes) aukletae, originally described from two species of auklets (Charadriiformes: Alcidae) from maritime eastern Russia, is reported from a third species of pelagic charadriiform (Stercorariidae), the pomarine jaeger, Stercorarius pomarinus (Temminck), from Florida. The specimens from the jaeger are slightly smaller, the genital apodeme is more heavily sclerotized, paired setae gm are twice as long and there are other minor variations in the idiosomal and leg chaetotaxy. These differences are not considered sufficient to warrant taxonomic separation at the species or subspecies level from the nominate species T. (A.) aukletae. The same hypopus occurring across different families of birds is unusual in the Hypoderatidae. The diversity in hosts from several orders of birds, low intensities of infection in the two species from Africa, low prevalences in alcids from Russia, and rarity of these hypoderatids in all surveyed hosts leads us to speculate that the true host affinities of species in the genus Thalassornectes are unknown. The alternative consideration is that these are simply uncommon species that are very host specific.

New records of subcutaneous mites (Acari:Hypoderatidae) in birds, with examples of potential host colonization events.

New host, geographic records, or both are established for 14 species of hypoderatid deutonymphs from 14 species of birds in North America. Ten of these records are regarded as examples of a potential host colonization event where these hypopi have become established in hosts other than those with which they are normally associated. Herein, potential host colonization events by hypoderatid deutonymphs are regarded as more of an ecologically determined than physiologically specific phenomenon, often specifically related to sharing of nesting sites in the same rookeries by different host taxa. Neottialges ibisicola Young & Pence is placed as a junior synonym of Neottialges plegadicola Fain. The taxonomic status of Hypodectes propus from columbid versus ardeid hosts needs further study.

Detection of Neospora caninum in tissue sections using a murine monoclonal antibody.

A murine monoclonal antibody (MAb 6G7), isotype IgG2a, produced against tachyzoites of Neospora caninum (isolate NC-1) reacted specifically with tachyzoites of N. caninum in an indirect immunofluorescent antibody test. MAb 6G7 did not react with tachyzoites of Toxoplasma gondii, sporozoites of Isospora suis, Eimeria bovis, or E. tenella, or merozoites of E. bovis in the indirect immunofluorescent antibody test. MAb 6G7 reacted positively with both tachyzoites and bradyzoites of N. caninum in an avidin-biotin peroxidase complex immunohistochemical test on formalin-fixed paraffin-embedded tissues. No reaction was observed with the following: tachyzoites and bradyzoites of T. gondii, T. gondii-like parasites, or Hammondia hammondi; bradyzoites of Frenkelia microti; schizonts and merozoites of Sarcocystis-like organisms; schizonts, sarcocysts, and oocysts/sporocysts of S. cruzi; schizonts and merozoites of S. canis; schizonts of S. hirsuta, S. tenella, and S. capracanis; merozoites of S. neurona and S. neurona-like organisms, E. bovis, or Haemoproteus sp.; bradyzoites and merozoites of S. montanaensis; bradyzoites of S. odocoileocanis, S. cruzi, and S. tenella; meronts, sexual stages, and caryocysts of Caryospora sp. and C. bigenetica; micromerozoites, macromerozoites, and schizonts of Hepatozoon canis; sporozoites, sexual stages, and oocysts of Cryptosporidium parvum and C. baileyi; trophozoites of Monocystis lumbrici, Tritrichomonas foetus, and Balantidium coli; tissue cysts and bradyzoites of Besnoitia sp. and B. jellisoni; amastigotes of Leishmania sp.; and trophic theronts of Ichthyopthirius multifilis. MAb 6G7 reacted with tachyzoites and bradyzoites of N. caninum in natural and experimental infections in dogs, cattle, mice, rats, sheep, and goats, indicating that host origin of the tissues did not affect the performance of the test.