Wavelength-dependent collagen fragmentation during mid-IR laser ablation

Mid-infrared free-electron lasers have proven adept in surgical applications. When tuned to wavelengths between 6 and 7 microm, such lasers remove defined volumes of soft tissue with very little collateral damage. Previous attempts to explain the wavelength-dependence of collateral damage have invoked a wavelength-dependent loss of protein structural integrity. However, the molecular nature of this structural failure has been heretofore ill-defined. In this report, we evaluate several candidates for the relevant transition by analyzing the nonvolatile debris ejected during ablation. Porcine corneas were ablated with a free-electron laser tuned to 2.77 or 6.45 microm-wavelengths with matched absorption coefficients for hydrated corneas that respectively target either tissue water or protein. The debris ejected during these ablations was characterized via gel electrophoresis, as well as Fourier transform infrared spectroscopy, micro-Raman and 13C-NMR spectroscopy. We find that high-fluence (240 J/cm2) ablation at 6.45 microm, but not at 2.77 microm, leads to protein fragmentation accompanied by the accumulation of nitrile and alkyne species. The candidate transition most consistent with these observations is scission of the collagen protein backbone at N-alkylamide bonds. Identifying this transition is a key step toward understanding the observed wavelength-dependence of collateral damage in mid-infrared laser ablation.     

The Pox in the North American Backyard: Volepox Virus Pathogenesis in California Mice (Peromyscus californicus)

Volepox virus (VPXV) was first isolated in 1985 from a hind foot scab of an otherwise healthy California vole (Microtus californicus). Subsequent surveys in San Mateo County, CA, revealed serological evidence suggesting that VPXV is endemic to this area, and a second viral isolate from a Pinyon mouse (Peromyscus truei) was collected in 1988. Since then, few studies have been conducted regarding the ecology, pathology, and pathogenicity of VPXV, and its prevalence and role as a potential zoonotic agent remain unknown. To increase our understanding of VPXV disease progression, we challenged 24 California mice (Peromyscus californicus) intranasally with 1.6×10(3) PFU of purified VPXV. By day five post infection (pi) we observed decreased activity level, conjunctivitis, ruffled hair, skin lesions, facial edema, and crusty noses. A mortality rate of 54% was noted by day eight pi. In addition, internal organ necrosis and hemorrhages were observed during necropsy of deceased or euthanized animals. Viral loads in tissues (brain, gonad, kidney, liver, lung, spleen, submandibular lymph node, and adrenal gland), bodily secretions (saliva, and tears), and excretions (urine, and/or feces) were evaluated and compared using real time-PCR and tissue culture. Viral loads measured as high as 2×10(9) PFU/mL in some organs. Our results suggest that VPXV can cause extreme morbidity and mortality within rodent populations sympatric with the known VPXV reservoirs.     

禽流感H5N1病毒感染BALB/c小鼠后多器官病变初探

为了研究禽流感H5N1病毒在各个器官的增殖和病理变化,在生物安全实验室,我们将禽流感H5N1病毒通过尾静脉接种BALB/C小鼠。结果小鼠在不经过适应的情况下,直接感染发病,甚至死亡。在观察的7天内,感染小鼠临床症状主要表现呼吸急促,体温、体重下降。尸检表现肺出血,心外膜坏死以及肝脏的坏死。组织病理检查表现心、肝、肺等多器官的病变。肺的病变伴有纤维化的弥漫性肺泡损伤;心肌外膜大量淋巴细胞浸润、坏死;肝细胞大量坏死,淋巴细胞浸润。心、肝的坏死病变在H5N1禽流感病毒相关的研究中未见报道。经过对各个组织器官的病毒载量的检测,未发现病毒在各个病变组织中的复制。免疫组化的检测,各个组织中也未检出阳性的细胞反应。因此,我们认为H5N1禽流感病毒感染小鼠引起多个器官组织的损伤,甚至死亡,不是病毒在器官的复制,而可能是病毒感染小鼠,产生炎症细胞因子的高度表达,损伤多个器官组织所致。     

N fate and transport under variable cropping history and fertilizer rate on loamy sand and clay loam soils: I. Calibration of the LEACHMN model

The need for efficient use of agricultural chemicals and their potential adverse impact on critical water resources have increased the use of simulation models of the soil and plant system. Nevertheless, there is currently little or no agreement concerning model validity and applicability in varied soils and environments. The research version of LEACHMN (the N subroutine of LEACHM) was calibrated using field data including soil physical, hydraulic, and chemical properties, and maize (Zea mays L.) N uptake collected from a 3-yr nitrate leaching experiment. The field site consisted of plot-size lysimeters on clay loam and loamy sand soils with N fertilizer rates of 22, 100 and 134 kg N ha^–1. The calibration involved adjusting nitrification, denitrification, and volatilization rate constants to optimize the fit between predicted and measured data. When calibrated for each treatment-year combination and soil type, the model simulations of soil profile NO₃–N distribution were generally successful. The N transformation rate constants yielded by the calibration efforts were similar or close to those used in other model simulation studies. At both sites, the calibrated rate constants for the first year (following sod plowdown) were different from those for the subsequent two years. Denitrification rate constants were consistently higher for the clay site than for the sand site, while the nitrification rate constants were lower. N rate of application appeared not to affect the rate constants within each year-site combination, suggesting that cropping history and soil type had the greatest effect on N transformation rates.     

N rate and transport under variable cropping history and fertilizer rate on loamy sand and clay loam soils: II. Performance of LEACHMN using different calibration scenarios

Testing of existing agronomic models is needed to ensure their validity and applicability to different soils, cropping systems and environments. Data collected from a 3-year field experiment of maize (zea mays L.) on a loamy sand and a clay loam soil were used to validate the research version of the LEACHMN model for water flow and N fate and transport. Three calibration scenarios with increasing levels of generalization for transformation rate coefficients were used based on: (i) each year, treatment and soil type (ii) 3-year average values for each treatment and soil type, and (iii) average over years and soil types. Model accuracy was tested using both graphical and statistical methods including 1:1 scale plot, root mean square error and normalized root mean square error, and correlation coefficient values. The model accurately predicted drainage water flow rate and volume under both sites. Calibrated N transformation rate constants for each treatment, year and soil type provided satisfactory predictions of growing season cumulative NO₃–N leaching losses, and accurate predictions of growing season cumulative maize N uptake at both sites. The use of 3-year average rate constant values for each site resulted in fairly satisfactory predictions of NO₃–N leaching losses on the clay loam site, but inaccurate predictions on the loamy sand site. The model provided accurate predictions of cumulative maize N uptake for both sites. Using the rate constant values averaged over years and soil types resulted mostly in inaccurate predictions. Use of year and soil type-specific N rate coefficients results in accurate LEACHMN predictions of N leaching and maize N uptake. When rate coefficients are generalized over years for each soil type, satisfactory model predictions may be expected when N dynamics are not strongly affected by yearly variations in organic N inputs.     

Variation in Resistance to High Hydrostatic Pressure and rpoS Heterogeneity in Natural Isolates of Escherichia coli O157:H7

Several natural isolates of Escherichia coli O157:H7 have previously been shown to exhibit stationary-phase-dependent variation in their resistance to inactivation by high hydrostatic pressure. In this report we demonstrate that loss of the stationary-phase-inducible sigma factor RpoS resulted in decreased resistance to pressure in E. coli O157:H7 and in a commensal strain. Furthermore, variation in the RpoS activity of the natural isolates of O157:H7 correlated with the pressure resistance of those strains. Heterogeneity was noted in the rpoS alleles of the natural isolates that may explain the differences in RpoS activity. These results are consistent with a role for rpoS in mediating resistance to high hydrostatic pressure in E. coli O157:H7.     

Time course of NADH oxidase, inducible nitric oxide synthase and peroxynitrite in diabetic retinopathy in the BBZ/WOR rat.

This study investigated the time course of NADH oxidase, a source of superoxide in the vascular endothelium, inducible nitric oxide synthase (iNOS), and peroxynitrite (ONOO(-)) in the BBZ/Wor rat, a spontaneous model of noninsulin dependent diabetes (NIDDM). Colloidal gold-labeled immunocytochemical studies of iNOS and nitrotyrosine, a marker for OONO(-), were done on sections of retinas from male BBZ/Wor rats in which NADH oxidase was localized by cerium derived cytochemistry at three time points: pre-diabetes (prior to the onset of hyperglycemia); new onset diabetes (2-6 days after onset of hyperglycemia); and chronic diabetes (4-18 months after onset of hyperglycemia). Control retinas were from age matched non-diabetic BB(DR)/Wor rats. The percentage of blood vessels positive for NADH oxidase increased significantly (P = 0.05) in new onset (64.2 +/- 6.5%) and chronic diabetes (83.2 +/- 11.4%), as compared to pre-diabetes (25.8 +/- 5.6%) and nondiabetic controls (33.6 +/- 15.9%). The percentage of blood vessels positive for iNOS immunoreactivity was significantly higher in new onset diabetic retinas (69.6 +/- 5.88%, P = 0.0001; 8.9 +/- 3.29 colloidal gold particles (cgp) /50 microm(2)) than in chronic diabetic retinas (49.9 +/- 9.75%; 7.9 +/- 5.12 cgp) and both were significantly higher (P = 0.0001) than in prediabetic (3.7 +/- 0.81%; 0.4 +/- 0.56 cgp) and nondiabetic control retinas (8.7 +/- 4.66%; 1.2 +/- 1.40 cgp). In new onset diabetes, levels of nitrotyrosine immunoreactivity (60.8 +/- 16.91 cgp) were significantly higher (P = 0.0001) than those in chronic diabetes (29.5 +/- 4.31 cgp); both were significantly higher (P = 0.0001) than those in prediabetic (8.2 +/- 1.70 cgp) and nondiabetic retinas (9.0 +/- 1.87 cgp). There was no cumulative increase in nitrotyrosine in the chronic diabetic retinas as a function of time. In rats with diabetes there was disruption of the inner blood-retinal barrier. These results suggest that iNOS and ONOO(-) may contribute to retinal damage in diabetes from the onset of hyperglycemia in NIDDM.     

An approach to practical persistence

 A basic problem in community ecology is determining whether a community of interacting species will survive in the long term. A criterion ensuring this is that of permanence (or uniform persistence), which is based on the idea that species densities for large time are above minimum non-zero levels. There are various mathematical techniques for investigating permanence, but they do not yield an estimate for the minimum levels, and these may lie below minimum viability levels in the biological sense of ‘Practical Persistence’. Here we study a technique for obtaining explicit expressions for the minimum levels when one species is ‘slow’. This is illustrated for a predator–prey problem governed by difference equations, and we note that the technique is applicable even when the dynamics is chaotic.. Received: 7 April 1997 / Revised version: 25 March 1998