Continuous precipitation-filtration process for initial capture of a monoclonal antibody product using a four-stage countercurrent hollow fiber membrane washing step.

The significant increase in product titers, coupled with the growing focus on continuous bioprocessing, has renewed interest in using precipitation as a low-cost alternative to Protein A chromatography for the primary capture of monoclonal antibody (mAb) products. In this work, a commercially relevant mAb was purified from clarified cell culture fluid using a tubular flow precipitation reactor with dewatering and washing provided by tangential flow microfiltration. The particle morphology was evaluated using an inline high-resolution optical probe, providing quantitative data on the particle size distribution throughout the precipitation process. Data were obtained in both a lab-built 2-stage countercurrent washing system and a commercial countercurrent contacting skid that provided 4 stages of continuous washing. The processes were operated continuously for 2 h with overall mAb yield of 92 ± 3% and DNA removal of nearly 3 logs in the 4-stage system. The high DNA clearance was achieved by selective redissolution of the mAb using a low pH acetate buffer. Host cell protein clearance was 0.59 ± 0.08 logs, comparable to that based on model predictions. The process mass intensity was slightly better than typical Protein A processes and could be significantly improved by preconcentration of the antibody feed material.

Effects and significance of groundwater for vegetation: A systematic review.

Groundwater represents a critical water source for plants, especially during drought, with continuous groundwater availability widely associated with the presence of ecological refugia and the preservation of biodiversity during periods of adverse conditions. Here, we present a systematic quantitative literature review of global groundwater and ecosystem interactions to synthesise current knowledge and identify key knowledge gaps and research priorities through a management lens. Despite increasing research on groundwater dependent vegetation since the late 1990s, significant geographical and ecological biases are evident with papers focused on arid regions or areas with significant anthropogenic changes. Of the 140 papers reviewed, desert and steepe arid landscapes accounted for 50.7 % and desert and xeric shrublands were represented in 37.9 % of papers. A third of papers (34.4 %) quantified groundwater uptake by ecosystems and groundwater contributions to transpiration, with studies examining the influence of groundwater on vegetation productivity, distribution, and composition also well represented. In contrast, groundwater influences on other ecosystem functions are relatively poorly explored. The research biases introduce uncertainty in the transferability of findings between locations and ecosystems limiting the generality of our current understanding. This synthesis contributes to consolidating a solid knowledge base of the hydrological and ecological interrelationships for managers, planners, and other decision-makers that is relevant to the landscapes and environments they manage, so can more effectively deliver ecological and conservation outcomes.

Number of Attractors in the Critical Kauffman Model Is Exponential.

The Kauffman model is the archetypal model of genetic computation. It highlights the importance of criticality, at which many biological systems seem poised. In a series of advances, researchers have honed in on how the number of attractors in the critical regime grows with network size. But a definitive answer has remained elusive. We prove that, for the critical Kauffman model with connectivity one, the number of attractors grows at least, and at most, as (2/sqrt[e])^{N}. This is the first proof that the number of attractors in a critical Kauffman model grows exponentially.

Sperm granulomas: Predictive factors and impacts on patency post vasectomy reversal.

The objective of this study was to identify factors that predict for sperm granuloma formation and the impact of sperm granuloma presence and quantity on vasectomy reversal (VR) outcomes. A cross sectional retrospective review of prospectively collected data, on the impact of granuloma on VR outcomes from a single academic center was performed. The impact of age, obstructive interval, intraoperative vasal fluid findings, anastomosis type, body mass index, tobacco use and total motile count (TMC) was determined. A total of 1550 men underwent VR between January 2000 and August 2019. Granulomas were present unilaterally in 23.3% (n = 361) and bilaterally in 14.2% (n = 220). On univariate analysis, increasing patient age negatively correlated with a larger number of granulomas (p = .011). Granuloma presence was associated with finding intact and motile sperm from the vasal stump intraoperatively (p = .001), and vasoepididymostomy anastomosis (p < .001). However, granuloma presence (and quantity) did not correlate with obstructive interval or maximum TMC. Tobacco use and body mass index (BMI) were not associated with granuloma presence. On multivariate analysis, granuloma quantity was not associated with TMC. Obstructive interval and vasovasostomy anastomosis were associated with higher TMC, while BMI was negatively associated with TMC. In conclusion, increasing age was negatively correlated with granuloma formation. Granuloma presence was associated with more favourable intraoperative fluid findings and anastomosis type, but not post-VR TMC, suggesting men with and without granulomas undergoing skilled microsurgery will have similar patency rates. Heavier men should be encouraged for weight loss prior to vasectomy reversal as increasing BMI was associated with lower TMC.

The Effect of Male Age Over 50 Years on Vasectomy Reversal Outcomes.

OBJECTIVE: Advancing paternal age is associated with impaired semen parameters. We sought to evaluate reproductive outcomes in men undergoing vasectomy reversal (VR) aged ≥50 vs <50 years. METHODS: Reproductive outcomes (obstructive interval, female age, anastomosis type, post-VR total motile count (TMC), and pregnancy) after VR were assessed for men aged <50 and ≥50 years. Statistical analysis was performed using Kruskal-Wallis rank sum or Chi-squared tests. Multiple logistic regression was used to identify factors associated with achieving pregnancy. RESULTS: A total of 2777 men <50 years and 353 men ≥50 years were included. The mean obstructive interval was 8.7 years less for men <50 years (8.9 vs 17.6 years, P <.001). The chances of needing a vasoepididymostomy were higher in men ≥50 years (19.5% vs 10.1%, P <.001). Post-VR total motile count was higher in men <50 years (59.3 vs 29.1 × 106/mL, P <.001). About 33.4% of men <50 years and 26.1% ≥50 years contributed to a pregnancy (P = .007). On multiple logistic regression analysis, obstructive interval <10 years (OR 1.295, P = .002) and female age <35 (OR 1.659, P <.001) were associated with achieving a pregnancy. A history of smoking was associated with decreased odds of achieving a pregnancy (OR 0.523, P <.001). Age <50 or ≥50 years at VR was not associated with achieving pregnancy (OR 0.852, P = 0.254). CONCLUSION: Compared to those ≥50 years, more men <50 years achieved a pregnancy after VR. However, on adjusted multivariable analysis, age at VR was not an independent predictor of achieving pregnancy. Shorter obstructive interval and female age were associated with achieving pregnancy, while a history of smoking was associated with decreased odds. Successful outcomes after VR can be achieved in older men, and VR should be considered in men ≥50 years, when performed by a trained microsurgeon.

Panmixia and Limited Interspecific Introgression in Coyotes (Canis latrans) from West Virginia and Virginia, USA.

The expansion of coyotes (Canis latrans) into the eastern United States has had major consequences for ecological communities and wildlife managers. Despite this, there has been little investigation of the genetics of coyotes across much of this region, especially outside of the northeast. Understanding patterns of genetic structure and interspecific introgression would provide insights into the colonization history of the species, its response to the modern environment, and interactions with other canids. We examined the genetic characteristics of 121 coyotes from the mid-Atlantic states of West Virginia and Virginia by genotyping 17 polymorphic nuclear DNA microsatellite loci. These genotypes were compared with those from other canid populations to evaluate the extent of genetic introgression. We conducted spatial clustering analyses and spatial autocorrelation to assess genetic structure among sampled coyotes. Coyotes across the 2 states had high genetic diversity, and we found no evidence of genetic structure. Six to sixteen percent of individuals displayed some evidence of genetic introgression from other species depending on the method and criteria used, but the population possessed predominantly coyote ancestry. Our findings suggested introgression from other canid populations has played less of a role in shaping the genetic character of coyotes in these states compared with populations closer to the Canadian border. Coyotes appear to display a panmictic population structure despite high habitat heterogeneity and heavy human influence in the spatial environment, underscoring the adaptability of the species.

Global versus local causes and health implications of high mercury concentrations in sharks from the east coast of South Africa.

Conservation concern regarding the overharvest of global shark populations for meat and fin consumption largely surrounds documented deleterious ecosystem effects, but may be further supported by improved knowledge of possibly high levels in their edible tissues (particularly meat) of the neurotoxin, methylmercury (CH3Hg). For many regions, however, little data exist on shark tissue Hg concentrations, and reasons for Hg variation within and among species or across regions are poorly understood. We quantified total Hg (THg) in 17 shark species (total n=283) from the east coast of South Africa, a top Hg emitter globally. Concentrations varied from means of around 0.1 mg kg(-1) dry weight (dw) THg in hardnose smoothhound (Mustelus mosis) and whale (Rhincodon typus) sharks to means of over 10 mg kg(-1) dw in shortfin mako (Isurus oxyrinchus), scalloped hammerhead (Sphyrna lewini), white (Carcharodon carcharias) and ragged-tooth (Carcharias taurus) sharks. These sharks had higher THg levels than conspecifics sampled from coastal waters of the North Atlantic and North, mid-, and South Pacific, and although sampling year and shark size may play a confounding role, this result suggests the potential importance of elevated local emissions. Values of THg showed strong, species-specific correlations with length, and nearly half the remaining variation was explained by trophic position (using nitrogen stable isotopes, δ(15)N), whereas measures of foraging habitat (using carbon stable isotopes, δ(13)C) were not significant. Mercury concentrations were above the regulatory guidelines for fish health effects and safe human consumption for 88% and 70% of species, respectively, suggesting on-going cause for concern for shark health, and human consumers of shark meat.

Comparative organochlorine accumulation in two ecologically similar shark species (Carcharodon carcharias and Carcharhinus obscurus) with divergent uptake based on different life history.

Trophic position and body mass are traits commonly used to predict organochlorine burdens. Sharks, however, have a variety of feeding and life history strategies and metabolize lipid uniquely. Because of this diversity, and the lipid-association of organochlorines, the dynamics of organochlorine accumulation in sharks may be predicted ineffectively by stable isotope-derived trophic position and body mass, as is typical for other taxa. The present study compared ontogenetic organochlorine profiles in the dusky shark (Carcharhinus obscurus) and white shark (Carcharodon carcharias), which differ in metabolic thermoregulation and trophic position throughout their ontogeny. Although greater organochlorine concentrations were observed in the larger bodied and higher trophic position white shark (e.g., p,p'-dichlorodiphenyldichloroethylene: 20.2 ± 2.7 ng/g vs 9.3 ± 2.2 ng/g in the dusky shark), slopes of growth-dilution corrected concentrations with age were equal to those of the dusky shark. Similar ontogenetic trophic position increases in both species, less frequent white shark seal predation than previously assumed, or inaccurate species-specific growth parameters are possible explanations. Inshore habitat use (indicated by δ(13)C values) and mass were important predictors in white and dusky sharks, respectively, of both overall compound profiles and select organochlorine concentrations. The present study clarified understanding of trophic position and body mass as reliable predictors of interspecific organochlorine accumulation in sharks, whereas regional endothermy and diet shifting were shown to have less impact on overall rates of accumulation.

Handheld Diffuse Reflectance Spectral Imaging (DRSi) for in-vivo characterization of skin.

Diffuse reflectance spectroscopy provides a noninvasive means to measure optical and physiological properties of tissues. To expand on these measurements, we have developed a handheld diffuse reflectance spectral imaging (DRSi) system capable of acquiring wide field hyperspectral images of tissue. The image acquisition time was approximately 50 seconds for a 50x50 pixel image. A transport model was used to fit each spectra for reduced scattering coefficient, hemoglobin concentration and melanin concentration resulting in optical property maps. The system was validated across biologically relevant levels of reduced scattering (5.14% error) and absorption (8.34% error) using tissue simulating phantoms. DRSi optical property maps of a pigmented skin lesion were acquired in vivo. These trends in optical properties were consistent with previous observations using point probe devices.

Effect of sample preparation techniques for carbon and nitrogen stable isotope analysis of hydroxyapatite structures in the form of elasmobranch vertebral centra.

RATIONALE: Bulk stable isotope analysis (SIA) provides an important tool for the study of animal ecology. Elasmobranch vertebral centra can be serially sampled to obtain an isotopic history of an individual over ontogeny. The measured total δ(13)C value, however, may be misinterpreted due to the inclusion of the (13)C-rich inorganic portion. Hydrochloric acid (HCl) is commonly used to remove the inorganic portion of hydroxyapatite structures before undertaking SIA, but more recently ethylenediaminetetraacetic acid (EDTA) has been recommended for elasmobranch vertebrae. These acid treatments may introduce uncertainty on measured δ(13)C and δ(15)N values above instrument precision and the effect of small sample size remains untested for elasmobranch vertebrae. METHODS: Using a non-dilution program on an isotope ratio mass spectrometer the minimum sample weight of vertebrae required to obtain accurate isotopic values was determined for three shark species: white (Carcharodon carcharias), tiger (Galeocerdo cuvier), and sand tiger (Carcharias taurus). To examine if acid treatment completely removes the inorganic component of the vertebrae or whether the technique introduces its own uncertainty on measured δ(13)C and δ(15)N values, vertebrae samples were analyzed untreated and following EDTA treatment. RESULTS: The minimum sample weight required for accurate stable isotope values and the percentage sample yield following EDTA treatment varied within and among species. After EDTA treatment, white shark vertebrae were all enriched in (13)C and depleted in (15) N, tiger shark vertebrae showed both enrichment and depletion of (13)C and (15)N, and sand tiger shark vertebrae were all depleted in (13)C and (15)N. CONCLUSIONS: EDTA treatment of elasmobranch vertebrae produces unpredictable effects (i.e. non-linear and non-correctable) among species in both the percentage sample yield and the measured δ(13)C and δ(15)N values. Prior to initiating a large-scale study, we strongly recommend investigating (i) the minimum weight of vertebral material required to obtain consistent isotopic values and (ii) the effects of EDTA treatment, specific to the study species and the isotope ratio mass spectrometer employed.

Rescaling the trophic structure of marine food webs.

Measures of trophic position (TP) are critical for understanding food web interactions and human-mediated ecosystem disturbance. Nitrogen stable isotopes (δ(15) N) provide a powerful tool to estimate TP but are limited by a pragmatic assumption that isotope discrimination is constant (change in δ(15) N between predator and prey, Δ(15) N = 3.4‰), resulting in an additive framework that omits known Δ(15) N variation. Through meta-analysis, we determine narrowing discrimination from an empirical linear relationship between experimental Δ(15) N and δ(15) N values of prey consumed. The resulting scaled Δ(15) N framework estimated reliable TPs of zooplanktivores to tertiary piscivores congruent with known feeding relationships that radically alters the conventional structure of marine food webs. Apex predator TP estimates were markedly higher than currently assumed by whole-ecosystem models, indicating perceived food webs have been truncated and species-interactions over simplified. The scaled Δ(15) N framework will greatly improve the accuracy of trophic estimates widely used in ecosystem-based management.

A coastal seawater temperature dataset for biogeographical studies: large biases between in situ and remotely-sensed data sets around the Coast of South Africa.

Gridded SST products developed particularly for offshore regions are increasingly being applied close to the coast for biogeographical applications. The purpose of this paper is to demonstrate the dangers of doing so through a comparison of reprocessed MODIS Terra and Pathfinder v5.2 SSTs, both at 4 km resolution, with instrumental in situ temperatures taken within 400 m from the coast. We report large biases of up to +6°C in places between satellite-derived and in situ climatological temperatures for 87 sites spanning the entire ca. 2 700 km of the South African coastline. Although biases are predominantly warm (i.e. the satellite SSTs being higher), smaller or even cold biases also appear in places, especially along the southern and western coasts of the country. We also demonstrate the presence of gradients in temperature biases along shore-normal transects - generally SSTs extracted close to the shore demonstrate a smaller bias with respect to the in situ temperatures. Contributing towards the magnitude of the biases are factors such as SST data source, proximity to the shore, the presence/absence of upwelling cells or coastal embayments. Despite the generally large biases, from a biogeographical perspective, species distribution retains a correlative relationship with underlying spatial patterns in SST, but in order to arrive at a causal understanding of the determinants of biogeographical patterns we suggest that in shallow, inshore marine habitats, temperature is best measured directly.

Development of a noncontact diffuse optical spectroscopy probe for measuring tissue optical properties.

Optical reflectance probes are often used as tools to obtain optical spectra from superficial tissues and subsequently determine optical and physiological properties associated with early stage cancer. These probes, when placed directly on the tissue, are known to cause significant pressure-dependent changes in local optical properties. To address this, we fit the probe with an optical device that images the illumination and collection fibers onto the tissue surface, eliminating the influence of contact probe pressure on the sampling area. The noncontact probe addition addresses new optical conditions that may affect its performance such as tissue surface contour, and specular reflections by implementing an autofocusing mechanism and cross polarization. Extracted optical properties of tissue simulating phantoms yield errors of 3.46% in reduced scattering and 8.62% in absorbance. Autofocusing has extended the depth of field from 4 mm to throughout the 12 mm range of autofocus travel, while cross polarization has removed the incidence angle dependent specular reflection component from the collected signal.

Default mode network connectivity in stable vs progressive mild cognitive impairment.

OBJECTIVE: Dysfunction of the default mode network (DMN) has been identified in prior cross-sectional fMRI studies of Alzheimer disease (AD) and mild cognitive impairment (MCI); however, no studies have examined its utility in predicting future cognitive decline. METHODS: fMRI scans during a face-name memory task were acquired from a cohort of 68 subjects (25 normal control, 31 MCI, and 12 AD). Subjects with MCI were followed for 2.4 years (±0.8) to determine progression to AD. Maps of DMN connectivity were compared with a template DMN map constructed from elderly normal controls to obtain goodness-of-fit (GOF) indices of DMN expression. Indices were compared between groups and correlated with cognitive decline. RESULTS: GOF indices were highest in normal controls, intermediate in MCI, and lowest in AD (p < 0.0001). In a predictive model (that included baseline GOF indices, age, education, Mini-Mental State Examination score, and an index of DMN gray matter volume), the effect of GOF index on progression from MCI to dementia was significant. In MCI, baseline GOF indices were correlated with change from baseline in functional status (Clinical Dementia Rating-sum of boxes) (r = -0.40, p < 0.04). However, there was no additional predictive value for DMN connectivity when baseline delayed recall was included in the models. CONCLUSIONS: fMRI connectivity indices distinguish patients with MCI who undergo cognitive decline and conversion to AD from those who remain stable over a 2- to 3-year follow-up period. Our data support the notion of different functional brain connectivity endophenotypes for "early" vs "late" MCI, which are associated with different baseline memory scores and different rates of progression and conversion.

Maternal investment and size-specific reproductive output in carcharhinid sharks.

1. Life-history theory predicts that organisms will provide an optimal level of parental investment for offspring survival balanced against the effects on their own survival and future reproductive potential. 2. Optimal resource allocation models also predict an increase in reproductive output with age as expected future reproductive effort decreases. To date, maternal investment in sharks has received limited attention. 3. We found that neonatal dusky sharks (Carcharhinus obscurus) are not independent from maternal resource allocation at the point of parturition but instead are provisioned with energy reserves in the form of an enlarged liver that constitutes approximately 20% of total body mass. 4. Analysis of long-term archived data sets showed that a large proportion of this enlarged liver is utilized during the first weeks or months of life suggesting that the reported weight loss of newborn sharks signifies a natural orientation process and is not necessarily related to prey abundance and/or indicative of high mortality rates. 5. Interrogation of near-term pup mass in two carcharhinids, the dusky and spinner shark (Carcharhinus brevipinna), further revealed an increase in reproductive output with maternal size, with evidence for a moderate decline in the largest mothers. 6. For the dusky shark, there was a trade-off between increasing litter size and near-term pup mass in support of optimal offspring size theory. 7. For both the dusky and spinner shark, there was a linear increase in near-term pup mass with month, which may indicate variable parturition strategies and/or that carcharhinids are able to adjust the length of the gestation period. 8. The identification of optimal size-specific reproductive output has direct implications for improving the reproductive potential of exploited shark populations and for structuring future management strategies.

Ammonia and carbon dioxide concentrations in disposable and reusable ventilated mouse cages.

This study compares resuable and disposable individually ventilated mouse cages in terms of the formation of intracage CO2 and NH3. Crl:CD-1(ICR) female mice were placed in either disposable or reusable ventilated cages in a positive pressure animal rack. Intracage CO2 and NH3 were measured once daily for 9 d; temperature and relative humidity were monitored for the first 7 d. Results indicated higher CO2 levels in the rear of the disposable cages and in the front of the reusable cages. This pattern corresponded to where the mice tended to congregate. However, CO2 concentrations did not differ significantly between the 2 cage types. Average CO2 levels in both cage types never exceeded approximately 3000 ppm. Intracage NH3 began to rise in the reusable cages on day 4, reached approximately 50 ppm by day 5 and by day 9 was greater than 150 ppm at the cages' rear sampling port while remaining at approximately 70 ppm at the front sampling port. Intracage NH3 levels in the disposable cages remained less than or equal to 3.2 ppm. Intracage temperature and relative humidity were approximately the same in both cage types. We concluded that the disposable ventilated cage performed satisfactorily under the conditions of the study.

State of security at US colleges and universities: a national stakeholder assessment and recommendations.

In 2004 the US Department of Justice, Office of Community Oriented Policing Services, sponsored a National Summit on Campus Public Safety. The summit brought together various stakeholders including campus police and security officials, local police chiefs, college and university faculty and administrators, federal officials, students and parents, and community leaders to address the issues and complexities of campus safety. Delegates to the summit identified key issues in campus safety and security, which included establishing a national center on campus safety, balancing traditional open environments with the need to secure vulnerable sites, improving coordination with state and local police, reducing internal fragmentation, elevating professionalism, and increasing eligibility of campus police and security agencies to compete for federal law enforcement funds. Focus on "active shooters" on campus, resulting from the Virginia Tech incident, should not diminish attention placed on the broader, more prevalent safety and security issues facing the nation's educational campuses. Recommendations resulting from the summit called for establishing a national agenda on campus safety, formation of a national center on campus public safety, and increased opportunity for campus police and security agencies to compete for federal and state funds.