Methodology for Robotic In Vitro Testing of the Knee.

The knee joint plays a pivotal role in mobility and stability during ambulatory and standing activities of daily living (ADL). Increased incidence of knee joint pathologies and resulting surgeries has led to a growing need to understand the kinematics and kinetics of the knee. In vivo, in silico, and in vitro testing domains provide researchers different avenues to explore the effects of surgical interactions on the knee. Recent hardware and software advancements have increased the flexibility of in vitro testing, opening further opportunities to answer clinical questions. This paper describes best practices for conducting in vitro knee biomechanical testing by providing guidelines for future research. Prior to beginning an in vitro knee study, the clinical question must be identified by the research and clinical teams to determine if in vitro testing is necessary to answer the question and serve as the gold standard for problem resolution. After determining the clinical question, a series of questions (What surgical or experimental conditions should be varied to answer the clinical question, what measurements are needed for each surgical or experimental condition, what loading conditions will generate the desired measurements, and do the loading conditions require muscle actuation?) must be discussed to help dictate the type of hardware and software necessary to adequately answer the clinical question. Hardware (type of robot, load cell, actuators, fixtures, motion capture, ancillary sensors) and software (type of coordinate systems used for kinematics and kinetics, type of control) can then be acquired to create a testing system tailored to the desired testing conditions. Study design and verification steps should be decided upon prior to testing to maintain the accuracy of the collected data. Collected data should be reported with any supplementary metrics (RMS error, dynamic statistics) that help illuminate the reported results. An example study comparing two different anterior cruciate ligament reconstruction techniques is provided to demonstrate the application of these guidelines. Adoption of these guidelines may allow for better interlaboratory result comparison to improve clinical outcomes.

Thermal Profiles of Chainsaw Hollows and Natural Hollows during Extreme Heat Events.

Loss of hollow-bearing trees threatens many hollow-dependent wildlife. To mitigate this process, artificial chainsaw-carved hollows (CHs) are often created in dead trees, yet little is known about their thermal profiles. We measured temperatures inside 13 natural hollows (8 live and 5 dead trees) and 45 CHs (5 live and 40 dead trees) in the central west of NSW, Australia, over the course of 2 summers. Maximum temperatures and daily temperature ranges within natural hollows and artificial hollows were similar in 2017-2018. Hollow temperatures were lower in thicker-walled hollows than in thinner-walled hollows. During the January 2019 heatwave, temperatures inside CHs in dead trees exceeded 4-35 °C higher than the upper limit of the thermal neutral zone of sugar gliders-for 6.2 consecutive days (range 0-9 days). CHs in dead trees provided little buffering from thermal extremes; when air temperatures peaked at 44.6 °C, CHs in dead trees were on average 2.4 °C cooler than ambient (range: 5.5 °C cooler to 1.0 °C hotter than ambient). These results show that CHs created in dead trees may not provide suitable thermal conditions for hollow-dependent marsupials during summer heatwaves. Retention of large live trees, coupled with revegetation, is crucial for conserving hollow-dependent fauna in agricultural landscapes.

Artificial refuges for wildlife conservation: what is the state of the science?

Artificial refuges are human-made structures that aim to create safe places for animals to breed, hibernate, or take shelter in lieu of natural refuges. Artificial refuges are used across the globe to mitigate the impacts of a variety of threats on wildlife, such as habitat loss and degradation. However, there is little understanding of the science underpinning artificial refuges, and what comprises best practice for artificial refuge design and implementation for wildlife conservation. We address this gap by undertaking a systematic review of the current state of artificial refuge research for the conservation of wildlife. We identified 224 studies of artificial refuges being implemented in the field to conserve wildlife species. The current literature on artificial refuges is dominated by studies of arboreal species, primarily birds and bats. Threatening processes addressed by artificial refuges were biological resource use (26%), invasive or problematic species (20%), and agriculture (15%), yet few studies examined artificial refuges specifically for threatened (Vulnerable, Endangered, or Critically Endangered) species (7%). Studies often reported the characteristics of artificial refuges (i.e. refuge size, construction materials; 87%) and surrounding vegetation (35%), but fewer studies measured the thermal properties of artificial refuges (18%), predator activity (17%), or food availability (3%). Almost all studies measured occupancy of the artificial refuges by target species (98%), and over half measured breeding activity (54%), whereas fewer included more detailed measures of fitness, such as breeding productivity (34%) or animal body condition (4%). Evaluating the benefits and impacts of artificial refuges requires sound experimental design, but only 39% of studies compared artificial refuges to experimental controls, and only 10% of studies used a before-after-control-impact (BACI) design. As a consequence, few studies of artificial refuges can determine their overall effect on individuals or populations. We outline a series of key steps in the design, implementation, and monitoring of artificial refuges that are required to avoid perverse outcomes and maximise the chances of achieving conservation objectives. This review highlights a clear need for increased rigour in studies of artificial refuges if they are to play an important role in wildlife conservation.

Increased Risk of Autopsy-Proven Pneumonia with Sex, Season and Neurodegenerative Disease.

There has been a markedly renewed interest in factors associated with pneumonia, a leading cause of death worldwide, due to its frequent concurrence with pandemics of influenza and Covid-19 disease. Reported predisposing factors to both bacterial pneumonia and pandemic viral lower respiratory infections are wintertime occurrence, older age, obesity, pre-existing cardiopulmonary conditions and diabetes. Also implicated are age-related neurodegenerative diseases that cause parkinsonism and dementia. We investigated the prevalence of autopsy-proven pneumonia in the Arizona Study of Aging and Neurodegenerative Disorders (AZSAND), a longitudinal clinicopathological study, between the years 2006 and 2019 and before the beginning of the Covid-19 pandemic. Of 691 subjects dying at advanced ages (mean 83.4), pneumonia was diagnosed postmortem in 343 (49.6%). There were 185 subjects without dementia or parkinsonism while clinicopathological diagnoses for the other subjects included 319 with Alzheimer's disease dementia, 127 with idiopathic Parkinson's disease, 72 with dementia with Lewy bodies, 49 with progressive supranuclear palsy and 78 with vascular dementia. Subjects with one or more of these neurodegenerative diseases all had higher pneumonia rates, ranging between 50 and 61%, as compared to those without dementia or parkinsonism (40%). In multivariable logistic regression models, male sex and a non-summer death both had independent contributions (ORs of 1.67 and 1.53) towards the presence of pneumonia at autopsy while the absence of parkinsonism or dementia was a significant negative predictor of pneumonia (OR 0.54). Male sex, dementia and parkinsonism may also be risk factors for Covid-19 pneumonia. The apolipoprotein E4 allele, as well as obesity, chronic obstructive pulmonary disease, diabetes, hypertension, congestive heart failure, cardiomegaly and cigarette smoking history, were not significantly associated with pneumonia, in contradistinction to what has been reported for Covid-19 disease.

Severe hyposmia distinguishes neuropathologically confirmed dementia with Lewy bodies from Alzheimer's disease dementia.

Many subjects with neuropathologically-confirmed dementia with Lewy bodies (DLB) are never diagnosed during life, instead being categorized as Alzheimer's disease dementia (ADD) or unspecified dementia. Unrecognized DLB therefore is a critical impediment to clinical studies and treatment trials of both ADD and DLB. There are studies that suggest that olfactory function tests may be able to distinguish DLB from ADD, but few of these had neuropathological confirmation of diagnosis. We compared University of Pennsylvania Smell Identification Test (UPSIT) results in 257 subjects that went on to autopsy and neuropathological examination. Consensus clinicopathological diagnostic criteria were used to define ADD and DLB, as well as Parkinson's disease with dementia (PDD), with (PDD+AD) or without (PDD-AD) concurrent AD; a group with ADD and Lewy body disease (LBD) not meeting criteria for DLB (ADLB) and a clinically normal control group were also included. The subjects with DLB, PDD+AD and PDD-AD all had lower (one-way ANOVA p < 0.0001, pairwise Bonferroni p < 0.05) first and mean UPSIT scores than the ADD, ADLB or control groups. For DLB subjects with first and mean UPSIT scores less than 20 and 17, respectively, Firth logistic regression analysis, adjusted for age, gender and mean MMSE score, conferred statistically significant odds ratios of 17.5 and 18.0 for the diagnosis, vs ADD. For other group comparisons (PDD+AD and PDD-AD vs ADD) and UPSIT cutoffs of 17, the same analyses resulted in odds ratios ranging from 16.3 to 31.6 (p < 0.0001). To our knowledge, this is the largest study to date comparing olfactory function in subjects with neuropathologically-confirmed LBD and ADD. Olfactory function testing may be a convenient and inexpensive strategy for enriching dementia studies or clinical trials with DLB subjects, or conversely, reducing the inclusion of DLB subjects in ADD studies or trials.

Cardiac sympathetic denervation and synucleinopathy in Alzheimer's disease with brain Lewy body disease.

Comorbid Lewy body pathology is very common in Alzheimer's disease and may confound clinical trial design, yet there is no in vivo test to identify patients with this. Tissue (and/or radioligand imaging) studies have shown cardiac sympathetic denervation in Parkinson's disease and dementia with Lewy bodies, but this has not been explored in Alzheimer's subjects with Lewy bodies not meeting dementia with Lewy bodies clinicopathological criteria. To determine if Alzheimer's disease with Lewy bodies subjects show sympathetic cardiac denervation, we analysed epicardial and myocardial tissue from autopsy-confirmed cases using tyrosine hydroxylase and neurofilament immunostaining. Comparison of tyrosine hydroxylase fibre density in 19 subjects with Alzheimer's disease/dementia with Lewy bodies, 20 Alzheimer's disease with Lewy bodies, 12 Alzheimer's disease subjects without Lewy body disease, 19 Parkinson's disease, 30 incidental Lewy body disease and 22 cognitively normal without Alzheimer's disease or Lewy body disease indicated a significant group difference (P < 0.01; Kruskal-Wallis analysis of variance) and subsequent pair-wise Mann-Whitney U tests showed that Parkinson's disease (P < 0.05) and Alzheimer's disease/dementia with Lewy bodies (P < 0.01) subjects, but not Alzheimer's disease with Lewy bodies subjects, had significantly reduced tyrosine hydroxylase fibre density as compared with cognitively normal. Both Parkinson's disease and Alzheimer's disease/dementia with Lewy bodies subjects also showed significant epicardial losses of neurofilament protein-immunoreactive nerve fibre densities within the fibre bundles as compared with cognitively normal subjects (P < 0.01) and both groups showed high pathologic alpha-synuclein densities (P < 0.0001). Cardiac alpha-synuclein densities correlated significantly with brain alpha-synuclein (P < 0.001), while cardiac tyrosine hydroxylase and neurofilament immunoreactive nerve fibre densities were negatively correlated with the densities of both brain and cardiac alpha-synuclein, as well as Unified Parkinson's Disease Rating Scale scores (P < 0.05). The clear separation of Alzheimer's disease/dementia with Lewy bodies subjects from Alzheimer's disease and cognitively normal, based on cardiac tyrosine hydroxylase fibre density, is the first report of a statistically significant difference between these groups. Our data do not show significant sympathetic cardiac denervation in Alzheimer's disease with Lewy bodies, but strongly confirm that cardiac nuclear imaging with a noradrenergic radioligand is worthy of further study as a potential means to separate Alzheimer's disease from Alzheimer's disease/dementia with Lewy bodies during life.

Prevalence of REM sleep behavior disorder in Sun City, Arizona.

OBJECTIVE: To determine prevalence of REM sleep behavior disorder (RBD) [prodromal Lewy body disease] in Sun City, Arizona. PATIENTS AND METHODS: We attempted, by telephone and mail, a survey using the RBD single item question for probable RBD (pRBD) and the Innsbruck RBD Inventory. Individuals answering "yes" to 4/5 Inventory questions were considered to have high likelihood RBD (HL-RBD.). RESULTS: Response rate was 484/3000 individuals contacted (16%), mean age 78; 48 (9.9%) endorsed pRBD by RBD1Q; 16 (3.3%) had HL-pRBD. Prevalence of idiopathic cases (without neurodegenerative disease) was 8.8% pRBD and 2.8% HL-RBD. CONCLUSION: Our estimated definite RBD prevalence of 1.7% (61.3% of HL-RBD) was similar to previous community-based studies.

Faster cognitive decline in dementia due to Alzheimer disease with clinically undiagnosed Lewy body disease.

BACKGROUND: Neuropathology has demonstrated a high rate of comorbid pathology in dementia due to Alzheimer's disease (ADD). The most common major comorbidity is Lewy body disease (LBD), either as dementia with Lewy bodies (AD-DLB) or Alzheimer's disease with Lewy bodies (AD-LB), the latter representing subjects with ADD and LBD not meeting neuropathological distribution and density thresholds for DLB. Although it has been established that ADD subjects with undifferentiated LBD have a more rapid cognitive decline than those with ADD alone, it is still unknown whether AD-LB subjects, who represent the majority of LBD and approximately one-third of all those with ADD, have a different clinical course. METHODS: Subjects with dementia included those with "pure" ADD (n = 137), AD-DLB (n = 64) and AD-LB (n = 114), all with two or more complete Mini Mental State Examinations (MMSE) and a full neuropathological examination. RESULTS: Linear mixed models assessing MMSE change showed that the AD-LB group had significantly greater decline compared to the ADD group (ß = -0.69, 95% CI: -1.05, -0.33, p<0.001) while the AD-DLB group did not (ß = -0.30, 95% CI: -0.73, 0.14, p = 0.18). Of those with AD-DLB and AD-LB, only 66% and 2.1%, respectively, had been diagnosed with LBD at any point during their clinical course. Compared with clinically-diagnosed AD-DLB subjects, those that were clinically undetected had significantly lower prevalences of parkinsonism (p = 0.046), visual hallucinations (p = 0.0008) and dream enactment behavior (0.013). CONCLUSIONS: The probable cause of LBD clinical detection failure is the lack of a sufficient set of characteristic core clinical features. Core DLB clinical features were not more common in AD-LB as compared to ADD. Clinical identification of ADD with LBD would allow stratified analyses of ADD clinical trials, potentially improving the probability of trial success.

Postharvest management of gray mold and brown rot on surfaces of peaches and grapes using electrolyzed oxidizing water.

This study evaluated the potential use of near-neutral (pH=6.3-6.5) electrolyzed oxidizing water (EO water) to inactivate pure cultures of Botrytis cinerea and Monilinia fructicola and to mitigate fungal infection of these organisms on fruit surfaces. Treatment of these organisms, in pure culture, with EO water at concentrations of 25, 50, 75, and 100ppm total residual chlorine (TRC) and 10 min of contact time resulted in a 6 log(10) spores/mL reduction of both organisms. A dip treatment or a dip and daily spray treatment of EO water were used to evaluate its ability to prevent or delay the onset of surface infection on fruit during postharvest packaging and in retail shelf environments. A 10 minute dip treatment of surface inoculated peaches (M. fructicola) in EO water prevented infection for 3 days and resulted in a 12.5% incidence of infection and a disease severity rating of 6% after 5 days of storage at 25 degrees C. Dipping of green table grapes inoculated with B. cinerea into EO water prevented infection for 7 days and resulted in a 1% incidence of infection and a disease severity rating of 2% after 10 days of storage at 25 degrees C. A dip and daily spray of peaches with EO water prevented infection for 12 days and resulted in a 10% incidence of infection and a 6% disease severity after 14 days of storage at 25 degrees C. A dip and daily spray of grapes with EO water prevented infection for 24 days and resulted in a 2% incidence of infection and a disease severity rating of 2% after 26 days of storage at 25 degrees C. The results from this study suggest that these solutions may prove to be effective for postharvest sanitation of fruit surfaces prior to packaging and may increase the shelf life of the fruit in commercial settings.

Reduction of bacteria on spinach, lettuce, and surfaces in food service areas using neutral electrolyzed oxidizing water.

Food safety issues and increases in food borne illnesses have promulgated the development of new sanitation methods to eliminate pathogenic organisms on foods and surfaces in food service areas. Electrolyzed oxidizing water (EO water) shows promise as an environmentally friendly broad spectrum microbial decontamination agent. EO water is generated by the passage of a dilute salt solution ( approximately 1% NaCl) through an electrochemical cell. This electrolytic process converts chloride ions and water molecules into chlorine oxidants (Cl(2), HOCl/ClO(-)). At a near-neutral pH (pH 6.3-6.5), the predominant chemical species is the highly biocidal hypochlorous acid species (HOCl) with the oxidation reduction potential (ORP) of the solution ranging from 800 to 900mV. The biocidal activity of near-neutral EO water was evaluated at 25 degrees C using pure cultures of Escherichia coli, Salmonella typhimurium, Staphylococcus aureus, Listeria monocytogenes, and Enterococcus faecalis. Treatment of these organisms, in pure culture, with EO water at concentrations of 20, 50, 100, and 120ppm total residual chlorine (TRC) and 10min of contact time resulted in 100% inactivation of all five organisms (reduction of 6.1-6.7log(10)CFU/mL). Spray treatment of surfaces in food service areas with EO water containing 278-310ppm TRC (pH 6.38) resulted in a 79-100% reduction of microbial growth. Dip (10min) treatment of spinach at 100 and 120ppm TRC resulted in a 4.0-5.0log(10)CFU/mL reduction of bacterial counts for all organisms tested. Dipping (10min) of lettuce at 100 and 120ppm TRC reduced bacterial counts of E. coli by 0.24-0.25log(10)CFU/mL and reduced all other organisms by 2.43-3.81log(10)CFU/mL.