Development and Validation of Indirect Enzyme-Linked Immunosorbent Assays for Detecting Antibodies to SARS-CoV-2 in Cattle, Swine, and Chicken.

Multiple domestic and wild animal species are susceptible to SARS-CoV-2 infection. Cattle and swine are susceptible to experimental SARS-CoV-2 infection. The unchecked transmission of SARS-CoV-2 in animal hosts could lead to virus adaptation and the emergence of novel variants. In addition, the spillover and subsequent adaptation of SARS-CoV-2 in livestock could significantly impact food security as well as animal and public health. Therefore, it is essential to monitor livestock species for SARS-CoV-2 spillover. We developed and optimized species-specific indirect ELISAs (iELISAs) to detect anti-SARS-CoV-2 antibodies in cattle, swine, and chickens using the spike protein receptor-binding domain (RBD) antigen. Serum samples collected prior to the COVID-19 pandemic were used to determine the cut-off threshold. RBD hyperimmunized sera from cattle (n = 3), swine (n = 6), and chicken (n = 3) were used as the positive controls. The iELISAs were evaluated compared to a live virus neutralization test using cattle (n = 150), swine (n = 150), and chicken (n = 150) serum samples collected during the COVID-19 pandemic. The iELISAs for cattle, swine, and chicken were found to have 100% sensitivity and specificity. These tools facilitate the surveillance that is necessary to quickly identify spillovers into the three most important agricultural species worldwide.

[Hyperosmolar Hyperglycemic State (HHS) with extreme hyperglycemia and hyperglycemia-induced hyponatremia in a patient with previously known diabetes mellitus type 2]. / Förväntad hyponatremi vid extrem hyperglykemi ­ begränsad evidens.

This case report covers a patient presenting unconscious to the emergency department with a serum glucose level of 125.6 mmol/L and an associated hyperglycemia-induced hyponatremia with a sodium level of 101 mmol/L. The glucose level is, to our knowledge, one of the most extreme values described in medical literature. Glucose and sodium levels were normalized following treatment with intravenous fluids, mainly Ringer's lactate solution and insulin infusion with an initial rate of 0,5 IU/kg/h. The patient needed ten days of hospital care, including five days at the intensive care unit. Shortly after discharge the patient had a complete recovery. However, the extreme values led to difficulties regarding correction rates, choice of treatment and monitoring, which is further discussed in the article.

Effects of Human Electro-Muscular Incapacitation (HEMI) Devices on Cardiovascular Changes in Anesthetized Swine as Measured by Transesophageal Echocardiography (TEE).

The abundance of, and reliance upon, human electro-muscular incapacitation (HEMI) devices, especially in law enforcement, has generated scrutiny and examination of these technologies. The purpose of this study was to examine cardiovascular effects resulting from typical (5 sec) and longer activation (20 sec) HEMI applications studying myocardial function and peripheral vascular system using a combination of invasive cardiovascular catheters and transesophageal echocardiography (TEE). Six healthy swine (Sus scrofa) 3-5 months in age and weighing between 60 and 86 kg were anesthetized and exposed to the TASER Model X26 waveform while transesophageal echocardiography was performed. Stroke volume was shown to statistically decrease during HEMI application indicating an increase in systemic vascular resistance, but HEMI application did not result in myocardial dysfunction ("cardiac stunning").

Transdermal Delivery of Enfuvirtide in a Porcine Model Using a Low-Frequency, Low-Power Ultrasound Transducer Patch.

Ultrasound-mediated transdermal delivery is a promising parenteral administration method for large-molecule or unstable medications. This study evaluated skin health and systemic delivery when administering enfuvirtide, an injectable anti-retroviral medication, over a 1-mo period in a porcine model using a low-frequency cymbal transducer. Three groups received twice-daily treatments: (i) enfuvirtide injection control (n = 12); (ii) saline ultrasound control (n = 6); and (iii) enfuvirtide ultrasound treatment (n = 13). Ultrasound parameters were as follows: 30-min exposure, 90 mW/cm², 24-26 kHz and 15% duty cycle. No statistical difference in trans-epidermal water loss, a measure of skin health and function, was seen between ultrasound-treated and control skin sites for either saline (p = 0.50) or enfuvirtide (p = 0.29) groups. Average trough plasma concentrations of enfuvirtide were 0.6 ± 0.2 and 2.8 ± 0.8 µg/mL for ultrasound and injection, respectively. Tolerability and efficacy results indicate that chronic, low-frequency ultrasound exposure can be a practical means for transdermal delivery of medications such as enfuvirtide.

Recovery of Native Renal Function in Patients with Hepatorenal Syndrome Following Combined Liver and Kidney Transplant with Mercaptoacetyltriglycine-3 Renogram: Developing a Methodology.

Many patients with hepatorenal syndrome (HRS) end up receiving a combined liver and kidney transplant (CKLT) with preservation of native kidneys, specially type 1 HRS since is characterizes by a very rapid deterioration of renal function. Eventually, most of the patients regain renal function, but it is unknown if this is due to the transplanted kidney, the recovery of native renal function, or both. The aim of this study is to evaluate if there is recovery of native renal function in patients with HRS following CKLT. 22 patients (16 men; 6 women) with history of HRS and status post CKLT were studied. Mercapto-acetyltriglycine-3 renograms in the anterior and posterior views with the three kidneys in the field of view were simultaneously acquired. The renograms were analyzed by creating regions of interest around the transplanted and native kidneys. Relative contribution to the renal function, clearance, and effective renal plasma flow for the transplanted and native kidneys were obtained. 1/22 (4.5%) patients presented with a very poor functioning transplanted kidney, in 15/22 (68%) cases the combined native renal function was markedly poorer than the transplanted renal function and in 6/22 (27%) native kidneys showed a contribution to the renal function similar to the transplanted kidney. In conclusion, our series show that around 32% of the HRS patients recovered their native renal function after CKLT. Identification of common factors that affect recovery of native renal function may help to avoid unnecessary renal transplants, significantly reducing morbidity and cost, while facilitating a reallocation of scarce donor resources.

A pipeline for completing bacterial genomes using in silico and wet lab approaches.

BACKGROUND: Despite the large volume of genome sequencing data produced by next-generation sequencing technologies and the highly sophisticated software dedicated to handling these types of data, gaps are commonly found in draft genome assemblies. The existence of gaps compromises our ability to take full advantage of the genome data. This study aims to identify a practical approach for biologists to complete their own genome assemblies using commonly available tools and resources. RESULTS: A pipeline was developed to assemble complete genomes primarily from the next generation sequencing (NGS) data. The input of the pipeline is paired-end Illumina sequence reads, and the output is a high quality complete genome sequence. The pipeline alternates the employment of computational and biological methods in seven steps. It combines the strengths of de novo assembly, reference-based assembly, customized programming, public databases utilization, and wet lab experimentation. The application of the pipeline is demonstrated by the completion of a bacterial genome, Thermotoga sp. strain RQ7, a hydrogen-producing strain. CONCLUSIONS: The developed pipeline provides an example of effective integration of computational and biological principles. It highlights the complementary roles that in silico and wet lab methodologies play in bioinformatical studies. The constituting principles and methods are applicable to similar studies on both prokaryotic and eukaryotic genomes.

The effects of continuous application of the TASER X26 waveform on Sus scrofa.

This study investigated and evaluated the safety margins of the continuous long duration (up to 30 min) effect of the TASER X26 waveform, using a Sus scrofa model. Long duration continuous stimulus has not been evaluated on humans or human surrogates prior to this study. Swine were used as models due to similarities with humans in their skin and cardiovascular systems. Very long duration was used to determine both exposure dose and possible adverse physiological effects of dose. The trial began with an application of 10 min, and subsequent animals received increasing exposure time up to a survived maximum duration of 30 min. At the onset of this work, it was hypothesized that there would be a time limit after which most animals would not survive consistent with increased dose response. However, this hypothesis was not supported by the experimental results. All animals (10 of 10) survived up to 3 min. Seven of the 10 animals survived up to a 10-min exposure and 3 of 5 animals with a 30-min target exposure survived the full exposure. Surviving animals were recovered and observed for 24 h, with no postrecovery deaths. This suggests that swine (based on physiology) will not experience a fatal event when exposed to the TASER X26 for a continuous 3 min. Conclusions regarding longer duration (10-30 min) are not as certain due to the small sample sizes at these time intervals.

Human electromuscular incapacitation devices characterization: a comparative study on stress and the physiological effects on swine.

Human electromuscular incapacitation devices or electromuscular disruption (EMD) devices are increasingly used in police and military applications. Most individuals who experience electromuscular incapacitation are in a stress-filled state, and the effects of prolonged or repeated exposures are not well understood. Three different commercially available EMD devices were tested randomly on 6 anesthetized pigs each for a total of 18 pigs. Each animal was exposed to an initial 60-second application of the EMD device as an initial stressor. The animals were then allowed to rest under anesthesia for 60 minutes followed immediately by a 180-second application of the same device. Arterial blood gases and serum samples were collected throughout the experiment to measure catecholamines (epinephrine, norepinephrine, and dopamine) and cortisol. All the devices produced some level of muscle tetany as a result of the electrical delivery to the animal. All the pigs showed a mixed metabolic and respiratory acidosis. Cortisol tended to decrease after the initial exposure and slightly increased over the rest period. The extreme muscular work caused by the electrical stimulation resulting in muscle contractions did not result in a strong stress response but did result in an immediate sympathetic response during both applications of the device leading to the conclusion that initial stressor followed by rest and prolonged EMD device application did not exhaust the sympathetic system. For healthy adult animals, despite the prolonged muscular exertion and physiological stress caused by EMD devices, the body should be able to mount an appropriate sympathetic response and recover normally.

Immune regulation of a chronic bacteria infection and consequences for pathogen transmission.

BACKGROUND: The role of host immunity has been recognized as not only playing a fundamental role in the interaction between the host and pathogen but also in influencing host infectiousness and the ability to shed pathogens. Despite the interest in this area of study, and the development of theoretical work on the immuno-epidemiology of infections, little is known about the immunological processes that influence pathogen shedding patterns. RESULTS: We used the respiratory bacterium Bordetella bronchiseptica and its common natural host, the rabbit, to examine the intensity and duration of oro-nasal bacteria shedding in relation to changes in the level of serum antibodies, blood cells, cytokine expression and number of bacteria colonies in the respiratory tract. Findings show that infected rabbits shed B. bronchiseptica by contact up to 4.5 months post infection. Shedding was positively affected by number of bacteria in the nasal cavity (CFU/g) but negatively influenced by serum IgG, which also contributed to the initial reduction of bacteria in the nasal cavity. Three main patterns of shedding were identified: i- bacteria were shed intermittently (46% of individuals), ii- bacteria shedding fell with the progression of the infection (31%) and iii- individuals never shed bacteria despite being infected (23%). Differences in the initial number of bacteria shed between the first two groups were associated with differences in the level of serum antibodies and white blood cells. These results suggest that the immunological conditions at the early stage of the infection may play a role in modulating the long term dynamics of B. bronchiseptica shedding. CONCLUSIONS: We propose that IgG influences the threshold of bacteria in the oro-nasal cavity which then affects the intensity and duration of individual shedding. In addition, we suggest that a threshold level of infection is required for shedding, below this value individuals never shed bacteria despite being infected. The mechanisms regulating these interactions are still obscure and more studies are needed to understand the persistence of bacteria in the upper respiratory tract and the processes controlling the intensity and duration of shedding.

Feasibility of in vivo transesophageal cardiac ablation using a phased ultrasound array.

Over 2.2 million Americans suffer from atrial fibrillation making it one of the most common arrhythmias. Cardiac ablation has shown a high rate of success in treating paroxysmal atrial fibrillation. Prevailing modalities for this treatment are catheter based radio-frequency ablation or surgery. However, there is measurable morbidity and significant costs and time associated with these invasive procedures. Due to these issues, developing a method that is less invasive to treat atrial fibrillation is needed. In the development of such a device, a transesophageal ultrasound applicator for cardiac ablation was designed, constructed and evaluated. A goal of this research was to create lesions in myocardial tissue using a phased array. Based on multiple factors from array simulations, transesophageal imaging devices and throat anatomy, a phased ultrasound transducer that can be inserted into the esophagus was designed and tested. In this research, a two-dimensional sparse phased array with the aperture size of 20.7 mm x 10.2 mm with flat tapered elements as a transesophageal ultrasound applicator was fabricated and evaluated with in vivo experiments. Five pigs were anesthetized; the array was passed through the esophagus and positioned over the heart. The array was operated for 8-15 min at 1.6 MHz with the acoustic intensity of 150-300 W/cm(2) resulting in both single and multiple lesions on atrial and ventricular myocardium. The average size of lesions was 5.1 +/- 2.1 mm in diameter and 7.8 +/- 2.5 mm in length. Based on the experimental results, the array delivered sufficient power to the focal point to produce ablation while not grossly damaging nearby tissue outside the target area. These results demonstrate a potential application of the ultrasound applicator to transesophageal cardiac surgery in atrial fibrillation treatment.

Noninvasive ultrasonic glucose sensing with large pigs (approximately 200 pounds) using a lightweight cymbal transducer array and biosensors.

BACKGROUND: To prevent complications in diabetes, the proper management of blood glucose levels is essential. Since conventional glucose meters require pricking fingers or other areas of the skin, a noninvasive method for monitoring blood glucose levels is desired. Using a lightweight cymbal transducer array, this study was conducted to noninvasively determine the glucose levels of pigs having a similar size to humans. METHOD: In vivo experiments using eight pigs (approximately 200 pounds) were performed in five groups. A cymbal array with four biosensors was attached to the axillary area of the pig. The array was operated at 20 kHz at special peak-temporal peak intensity (I(sptp)) equal to 50 or 100 mW/cm(2) for 5, 10, or 20 minutes. After the ultrasound exposure, glucose concentrations of the interstitial fluid were determined using biosensors. For comparison, glucose levels of blood samples collected from the ear vein were measured by a commercial glucose meter. RESULT: In comparison, glucose levels determined by a cymbal array and biosensor system were close to those measured by a glucose meter. After a 20-minute ultrasound exposure at I(sptp) = 100 mW/cm(2), the average glucose level determined by the ultrasound system was 175 +/- 7 mg/dl, which is close to 166 +/- 5 mg/dl measured by the glucose meter. CONCLUSION: Results indicate the feasibility of using a cymbal array for noninvasive glucose sensing on pigs having a similar size to humans. Further studies on the ultrasound conditions, such as frequency, intensity, and exposure time, will be continued for effective glucose sensing.

Ultrasound mediated transdermal insulin delivery in pigs using a lightweight transducer.

PURPOSE: In previous studies, ultrasound mediated transdermal drug delivery has shown a promising potential as a method for noninvasive drug administration. For prospective future human application, this study was designed to determine the feasibility of lightweight cymbal transducer array as a practical device for noninvasive transdermal insulin delivery in large pigs. MATERIALS AND METHODS: Six Yorkshire pigs (100-140 lbs) were divided into two groups. As the control (n = 3), the first group did not receive any ultrasound exposure with the insulin. The second group (n = 3) was treated with ultrasound and insulin at 20 kHz with an I(sptp) = 100 mW/cm(2) at a 20% duty cycle for 60 min. With the pigs in lateral recumbency after anesthesia, the ultrasound transducer with insulin was placed on the axillary area of the pig. At the beginning and every 15 min up to 90 min, the blood glucose level was determined using a glucose monitoring system. To compare the results of individual animals, the change of blood glucose level was normalized to each animal's initial glucose value at the start of the experiment. RESULTS: Although each animal had a different initial glucose level, the mean and standard error for the six animals was 146 +/- 13 mg/dl. For the control group, the blood glucose level increased to 31 +/- 21 mg/dl compared to the initial baseline over the 90 min experiment. However for the ultrasound with insulin treated group, the glucose level decreased to -72 +/- 5 mg/dl at 60 min (p < 0.05) and continued to decrease to -91 +/- 23 mg/dl in 90 min (p < 0.05). CONCLUSION: The results indicate the feasibility of ultrasound mediated transdermal insulin delivery using the cymbal transducer array in animal with a similar size and weight to a human. Based on these result, the cymbal array has potential as a practical ultrasound system for noninvasive transdermal insulin delivery for diabetes management.