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BACKGROUNDCOVID-19 convalescent plasma (CCP) virus-specific antibody levels that translate into recipient posttransfusion antibody levels sufficient to prevent disease progression are not defined.METHODSThis secondary analysis correlated donor and recipient antibody levels to hospitalization risk among unvaccinated, seronegative CCP recipients within the outpatient, double-blind, randomized clinical trial that compared CCP to control plasma. The majority of COVID-19 CCP arm hospitalizations (15/17, 88%) occurred in this unvaccinated, seronegative subgroup. A functional cutoff to delineate recipient high versus low posttransfusion antibody levels was established by 2 methods: (i) analyzing virus neutralization-equivalent anti-Spike receptor-binding domain immunoglobulin G (anti-S-RBD IgG) responses in donors or (ii) receiver operating characteristic (ROC) curve analysis.RESULTSSARS-CoV-2 anti-S-RBD IgG antibody was volume diluted 21.3-fold into posttransfusion seronegative recipients from matched donor units. Virus-specific antibody delivered was approximately 1.2 mg. The high-antibody recipients transfused early (symptom onset within 5 days) had no hospitalizations. A CCP-recipient analysis for antibody thresholds correlated to reduced hospitalizations found a statistical significant association between early transfusion and high antibodies versus all other CCP recipients (or control plasma), with antibody cutoffs established by both methods-donor-based virus neutralization cutoffs in posttransfusion recipients (0/85 [0%] versus 15/276 [5.6%]; P = 0.03) or ROC-based cutoff (0/94 [0%] versus 15/267 [5.4%]; P = 0.01).CONCLUSIONIn unvaccinated, seronegative CCP recipients, early transfusion of plasma units in the upper 30% of study donors antibody levels reduced outpatient hospitalizations. High antibody level plasma units, given early, should be reserved for therapeutic use.TRIAL REGISTRATIONClinicalTrials.gov NCT04373460.FUNDINGDepartment of Defense (W911QY2090012); Defense Health Agency; Bloomberg Philanthropies; the State of Maryland; NIH (3R01AI152078-01S1, U24TR001609-S3, 1K23HL151826NIH); the Mental Wellness Foundation; the Moriah Fund; Octapharma; the Healthnetwork Foundation; the Shear Family Foundation; the NorthShore Research Institute; and the Rice Foundation.

Background/Objective: Ectopic cosecretion of corticotropin-releasing hormone (CRH) and adrenocorticotropic hormone (ACTH) in silent (ie, non–catecholamine-secreting) pheochromocytoma is a rare cause of Cushing syndrome. Case Report: A 57-year-old woman rapidly developed hypercortisolism, clinically manifesting as fatigue, muscle weakness, weight gain, and worsening hypertension and biochemically characterized by hypokalemia and marked increases in the serum cortisol and plasma ACTH levels. This acute presentation suggested a diagnosis of ectopic ACTH syndrome (EAS). Imaging studies revealed a right adrenal mass that enhanced after administration of the radioisotope gallium-68-DOTATATE. Plasma metanephrines were normal in 2 separate measurements. The possibility of a silent pheochromocytoma was considered. After controlling her hypercortisolism with metyrapone and surgical preparation with alpha blockade, the patient underwent elective right adrenalectomy. Pathology revealed a pheochromocytoma that stained focally for ACTH and CRH. Postoperatively, the cortisol levels normalized, the hypothalamic-pituitary-adrenal axis was not suppressed, and clinical symptoms from hypercortisolism abated. Discussion: Patients who exhibit a rapid progression of ACTH-dependent hypercortisolism should be screened for EAS. The use of functional imaging radioisotopes (eg, gallium DOTA-peptides) improves the detection of ACTH-secreting tumors. Preoperative treatment with steroidogenesis inhibitors helps control clinical and metabolic derangements associated with severe hypercortisolemia, whereas alpha blockade prevents the onset of an adrenergic crisis. Conclusion: We present a rare case of EAS due to a silent pheochromocytoma that cosecreted ACTH and CRH. Pheochromocytoma should be considered in patients with EAS who have an adrenal mass even in the absence of excessive catecholamine secretion.

Turfgrass systems hold significant climate change mitigation value, but their management often negates the beneficial effects due to the intense adoption of external inputs. The research objective in this paper was to assess the nitrogen fertilization rate able to maintain the ideal esthetic characteristics of Zoysia turfgrass, reducing the environmental impacts associated with greenhouse gas (GHG) emissions. A two-year open field experiment was conducted. Nitrogen was added to the soil at six rates (0, 50, 100, 150, 200, and 250 kg ha−1). The GHG emissions were monitored using a portable gas analyzer and the static chamber methodology. Cumulative environmental impacts were calculated from the inclusion of CO2, CH4, and, N2O using the Global Warming Potential (GWP). The quality assessment of the turf was assessed through a visual and instrumental approach. Higher CO2 and N2O fluxes were linked to high nitrogen rates, ranging from 83.55 to 87.50 and from 0.046 to 0.047 g N-N2O ha−1 day−1 for 200 and 250 kg N ha−1, respectively. CH4 emissions were not correlated to nitrogen rates. Higher GWP impacts were linked to high N rate treatments. A rate of 100 kg N ha−1 is recommended as the best strategy to reduce GHG emissions while maintaining high turf quality.

ABSTRACT Natural infection with the SARS-CoV-2 virus and vaccination increase the serum spike antibody levels. These levels, which differ among the various commercial assays, are used by the FDA to qualify individuals as potential COVID-19 convalescent plasma (CCP) donors. Over a 3-year period (April 2020–February 2023), we performed a retrospective analysis of spike IgG antibodies measured by a tertiary hospital clinical immunology laboratory using the Euroimmun SARS-CoV-2 IgG ELISA. The 3-year interval was classified into five periods based on the SARS-CoV-2 strain variant epidemiology. A total of 15,820 sera, derived from 11,022 individuals (6,362 females and 4,660 males) ranging from severe immunocompromised state to routine health visits, were tested. Spike IgG levels rose significantly over time, from a median ELISA 0.13 Arbitrary Units (AU) in the first period to 48.7 in the last period (P < 0.0001). The 80th percentile of the spike IgG distribution was 0.55, 8.1, 9.6, 64.9, and 151 AU in the five periods. Using 3.5 AU, the FDA threshold for qualifying CCP donors with this Euroimmun assay, the percentage of subjects eligible for CCP donation would have been 11%, 44%, 61%, 81%, and 91% in the five time periods. Overall, spike antibody levels have risen more than 100-fold during the pandemic, while SARS-CoV-2 variants have become resistant to monoclonal antibodies. Since CCP containing high titers of spike antibodies is known to be most effective against variants, restricting CCP donors to those with antibody values in the upper two deciles may allow greater therapeutic transfusion protection. IMPORTANCE Despite the evolution of SARS-CoV-2 variants of concern and ongoing transmission, COVID-19 hospitalization and mortality rates continue to decline. Both the percent seropositive and antibody levels have risen over the past 3 years. Here, we observe more than 90% seropositivity as well as more than a hundred-fold increase in spike IgG levels in a tertiary hospital clinical immunology laboratory setting. Antibody effector functions (such as neutralization, opsonization, and complement activation) and cell-mediated immunity all contribute to protection from COVID-19 progression to hospitalization, and all correlate to the total SARS-CoV-2 antibody levels. We recommend therapeutic COVID-19 convalescent plasma be restricted to the top 20% of potential donors to maintain activity against ongoing SARS-CoV-2 variant evolution.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of an ongoing pandemic that has infected over 36 million and killed over 1 million people. Informed implementation of government public health policies depends on accurate data on SARS-CoV-2 immunity at a population scale. We hypothesized that detection of SARS-CoV-2 salivary antibodies could serve as a noninvasive alternative to serological testing for monitoring of SARS-CoV-2 infection and seropositivity at a population scale. We developed a multiplex SARS-CoV-2 antibody immunoassay based on Luminex technology that comprised 12 CoV antigens, mostly derived from SARS-CoV-2 nucleocapsid (N) and spike (S). Saliva and sera collected from confirmed coronavirus disease 2019 (COVID-19) cases and from the pre-COVID-19 era were tested for IgG, IgA, and IgM to the antigen panel. Matched saliva and serum IgG responses (n = 28) were significantly correlated. The salivary anti-N IgG response resulted in the highest sensitivity (100%), exhibiting a positive response in 24/24 reverse transcription-PCR (RT-PCR)-confirmed COVID-19 cases sampled at >14 days post-symptom onset (DPSO), whereas the salivary anti-receptor binding domain (RBD) IgG response yielded 100% specificity. Temporal kinetics of IgG in saliva were consistent with those observed in blood and indicated that most individuals seroconvert at around 10 DPSO. Algorithms employing a combination of the IgG responses to N and S antigens result in high diagnostic accuracy (100%) by as early as 10 DPSO. These results support the use of saliva-based antibody testing as a noninvasive and scalable alternative to blood-based antibody testing.

Gaining information on the impact of lawn management with autonomous mowers on the floristic composition is crucial to improve their plant biodiversity. In this study, an autonomous mower with a reduced mowing frequency and a more sporadic mowing management system with a ride-on rotary mower were compared in terms of the effect on three dicotyledonous species (Phyla nodiflora, Lotus corniculatus and Sulla coronaria) transplanted onto stands of Bermuda and Manila grass. Regardless of the management system, P. nodiflora achieved the best results in terms of survival for both lawns (74.92 and 58.57% in Manila and Bermuda grass, respectively). In Bermuda grass, a higher percentage of surviving individuals was observed for the ordinary mower management system (42.59%), rather than with the autonomous mower (9.10%), while no differences emerged on Manila grass. On both Manila and Bermuda grass, a higher average percentage of coverage for single individual was observed for the ordinary mower management system (1.60 and 0.37%, respectively) compared to the autonomous mower system (0.55 and 0.08%, respectively). P. nodiflora had a higher percentage of individuals with flowers with the ordinary management system rather than with autonomous mower system both on Manila (60.73% and 33.90%, respectively) and Bermuda grass (48.66 and 3.32%, respectively). Despite a lower impact on the planted species being observed for the ordinary mower management system, encouraging results were obtained with the autonomous mower, for instance regarding the percentage of surviving individuals for P. nodiflora (33.95%) and L. corniculatus (22.08%) on Bermuda grass and the percentage of individuals with flowers for the same two species (33.90 and 13.59%, respectively) on Manila grass. Furthermore, the autonomous mower management system’s primary energy consumption over the year was lower compared to that of the ordinary system both on Manila (200.4 and 614.97 kWh ha−1 year−1, respectively) and Bermuda grass (177.82 and 510.99 kWh ha−1 year−1, respectively).

BackgroundVeno-venous extracorporeal membrane oxygenation (VV-ECMO) has been used in patients with COVID-19 acute respiratory distress syndrome (ARDS). We aim to assess the characteristics of delirium and describe its association with sedation and in-hospital mortality.MethodsWe retrospectively reviewed adult patients on VV-ECMO for severe COVID-19 ARDS in the Johns Hopkins Hospital ECMO registry in 2020–2021. Delirium was assessed by the Confusion Assessment Method for the ICU (CAM-ICU) when patients scored−3 or above on the Richmond Agitation-Sedation Scale (RASS). Primary outcomes were delirium prevalence and duration in the proportion of days on VV-ECMO.ResultsOf 47 patients (median age = 51), 6 were in a persistent coma and 40 of the remaining 41 patients (98%) had ICU delirium. Delirium in the survivors (n = 21) and non-survivors (n = 26) was first detected at a similar time point (VV-ECMO day 9.5(5,14) vs. 8.5(5,21), p = 0.56) with similar total delirium days on VV-ECMO (9.5[3.3, 16.8] vs. 9.0[4.3, 28.3] days, p = 0.43). Non-survivors had numerically lower RASS scores on VV-ECMO days (−3.72[−4.42, −2.96] vs. −3.10[−3.91, −2.21], p = 0.06) and significantly prolonged delirium-unassessable days on VV-ECMO with a RASS of −4/−5 (23.0[16.3, 38.3] vs. 17.0(6,23), p = 0.03), and total VV-ECMO days (44.5[20.5, 74.3] vs. 27.0[21, 38], p = 0.04). The proportion of delirium-present days correlated with RASS (r = 0.64, p < 0.001), the proportions of days on VV-ECMO with a neuromuscular blocker (r = −0.59, p = 0.001), and with delirium-unassessable exams (r = −0.69, p < 0.001) but not with overall ECMO duration (r = 0.01, p = 0.96). The average daily dosage of delirium-related medications on ECMO days did not differ significantly. On an exploratory multivariable logistic regression, the proportion of delirium days was not associated with mortality.ConclusionLonger duration of delirium was associated with lighter sedation and shorter paralysis, but it did not discern in-hospital mortality. Future studies should evaluate analgosedation and paralytic strategies to optimize delirium, sedation level, and outcomes.

Several trials have been carried out by various authors concerning autonomous mowers, which are battery-powered machines. The effects of these machines on turfgrass quality and energy consumption have been thoroughly investigated. However, there are still some aspects that have not been studied. Among these, random trajectory overlapping is one of the most important. To investigate these aspects, two RTK-GPS devices along with the custom-built software used for previous trials has been upgraded in order to precisely calculate how many times the mower drives over the same spot using random trajectories. This parameter, the number of passages in the same position, was hypothesized to explain the autonomous mower’s overlapping and trampling action. The trial has been carried out testing a commercial autonomous mower on three areas with different levels of complexity to assess its performances. The following variables were examined: the percentage of mowed area, the distance travelled, the number of intersections, the number of passages, and the autonomous mower’s work efficiency. The average percentage of area mown (average value for the three areas) was 54.64% after one hour and 80.15% after two hours of work. Percentage of area mown was 15% higher for the area with no obstacles after two hours of work. The number of passages was slightly different among the three garden designs. The garden with no obstacles obtained the highest number of passages with an average of 37 passages. The highest working efficiency was obtained in the garden with an intermediate number of obstacles with a value of 0.40 after two hours of work. The estimated energy consumption resulted 0.31 Wh m−2 after one hour and 0.42 Wh m−2 after two hours of working. These results highlight how the correct settings of cutting time may be crucial to consistently save energy during the long period and may be useful for a complete automation of the maintenance of green areas.

Objectives: The objectives were to characterize the liver kidney microsome (LKM) antibody profile of a 14-month-old girl with autoimmune hepatitis and analyze the laboratory prevalence of LKM positivity. Design and methods: This is retrospective analysis of the LKM antibody immunofluorescence tests performed by the Immunology Laboratory of Johns Hopkins Hospital from September 8, 2020 to July 31, 2022. LKM positive sera were also tested by an ELISA for LKM1 antibodies, which recognize the cytochrome P450 2D6 antigen. In silico analysis of 2D6 mRNA expression across anatomical sites was performed using Bgee and GTEx Portal databases. Results: Of the total of 1598 patients (893 F, 705 M, ages 0.8–94 years) tested for LKM antibodies, 3 were positive, yielding a 0.2% period prevalence. The clinical diagnosis was autoimmune hepatitis in the index case, acute viral hepatitis in a 3-yo male, and hepatocellular carcinoma in a 54-yo male. LKM antibodies yielded the classical homogenous staining pattern in the liver cytosol and proximal kidney tubular cells. The first two patients were also positive for LKM1 antibodies, whereas the third was negative. 2D6 mRNA was expressed highly in the liver, moderately in the duodenum, and minimally in other tissues. Conclusions: Overall, LKM antibodies are rare. They contribute to establish a diagnosis of autoimmune hepatitis, although they are also found in other liver diseases. The cytochrome P450 2D6 is one of the antigens recognized by LKM antibodies, but other antigens are likely targeted considering that 2D6 is minimally expressed in the kidney and yet LKM antibodies bind to kidney tubuli.

Background The response of solid tumors such as papillary thyroid cancer (PTC) to immune checkpoint inhibitors (ICIs) is highly variable. The biological basis of this variability remains unknown.Methods To test the hypothesis that preconditioning of the immune system modulates the therapeutic effect of ICIs, we used a murine model where PTC and iodine exacerbated thyroiditis (IET) can be induced in a temporally predictable fashion. A total of 122 mice were divided into 3 experimental groups. In the first one, named concomitant IET and PTC (No.=40), IET, and PTC were induced at the same time; in the second one, named pre-existing IET (No.=44), IET was induced prior to the induction of PTC; in the third one, named no IET (No.=38), only PTC was induced. Following disease induction, mice of each group were treated with anti-PD-1 antibody, anti-lymphocyte activation gene 3 antibody (anti-Lag3), anti-T-cell immunoglobulin and mucin domain 3 antibody (anti-Tim3), or IgG control. Ten weeks after the initial ICI injection, mice were sacrificed to collect the thyroid gland for histological analysis, to quantify the incidence and burden of PTC, and to perform high-throughput single-cell RNA sequencing of infiltrating CD45+ cells.Results In the concomitant IET and PTC group, ICI treatment reduced PTC incidence (p=0.002 comparing treatment with any ICI vs control), while it had no effect in the pre-existing IET and no IET groups. Single-cell sequencing of thyroidal CD45+ cells showed that the different ICIs tested had both specific and shared effects on all the components of the thyroidal immune cell infiltrate. The shared effect of the tested ICIs was dependent on the presence of pre-existing versus concomitant IET. In the context of concomitant IET, ICI treatment resulted in the modulation of a greater number of pathways related to both innate and adaptive immunity.Conclusions Response to ICIs depends on the status of the immune system of the treated individual. Modulation of the immune system should be explored as a tool to improve response to ICIs in patients with PTC or other forms of cancer.

ABSTRACT SARS-CoV-2 antibody levels wane following two-doses of mRNA vaccination. An mRNA booster dose provides increased protection against hospitalization and death. We demonstrated that a booster dose provides a significant increase in the neutralization of the Beta, Delta and Omicron variants in addition to an increased neutralization of the vaccine strain. The total spike IgG measurements, obtained by using commercial kits that target the spike protein from the vaccine strain, may not reflect serum neutralization against variants of concern. IMPORTANCE This study found little to no neutralizing capability following a 2-dose mRNA vaccine series against the omicron variant, and neutralizing capacity to any variant strain tested was lost by 8-months post 2-dose series. However, the mRNA booster dose eliminated the immune escape observed by the Omicron variant, following the 2-dose series. Even more, the neutralizing titers were significantly higher for all variants post-boost, compared to the titers from the post-two-dose series. Our data are unique, using paired samples that eliminate potential confounders that may impact vaccine response. Notably, as seen after the primary two-dose vaccine series, total antibody levels did not correlate perfectly with variant neutralization activity, suggesting that simply testing titers as a measure of protection may not be a long-term solution. Therefore, it is important to reassess the utility of SARS-CoV-2 antibody testing, as current vaccine strain-based testing may not reliably detect reactive antibodies to Omicron or other variants of concern.

Robotic solutions and technological advances for turf management demonstrated excellent results in terms of quality, energy, and time consumption. Two battery-powered autonomous mowers (2 WD and 4 WD) with random patterns were evaluated according to different trampling levels (control, low, medium, high) on a typical warm season turfgrass at the DAFE, University of Pisa, Italy. Data on the percentage of area mowed, the distance traveled, the number of passages, and the number of intersections were collected through RTK devices and processed by a custom-built software (1.8.0.0). The main quality parameters of the turfgrass were also analyzed by visual and instrumental assessments. Soil penetration resistance was measured through a digital penetrometer. The efficiency significantly decreased as the trampling level increased (from 0.29 to 0.11). The over-trampled areas were mainly detected by the edges (on average for the medium level: 18 passages for the edges vs. 14 in the central area). The trampling activity caused a reduction in turf height (from about 2.2 cm to about 1.5 cm). The energy consumption was low and varied from 0.0047 to 0.048 kWh per cutting session. Results from this trial demonstrated suitable quality for a residential turf of the Mediterranean area (NDVI values from 0.5 to 0.6), despite the over-trampling activity. Soil penetration data were low due to the reduced weight of the machines, but slightly higher for the 4 WD model (at 5 cm of depth, about 802 kPa vs. 670 kPa).

Background: The small intestine is one of the most sensitive organs to ischemia. L-arginine has been shown to reduce damage from ischemia and reperfusion injury. We hypothesize that explanted intestinal segments from rats will demonstrate reduced susceptibility to ischemic injury when perfused with l-arginine. Methods: 45 small intestinal segments were harvested from male Sprague-Dawley rats and connected to an ex vivo intestinal perfusion device. Ischemic damage was induced by perfusing the extraluminal side with Ringer-HEPES buffer saturated with 100% N2. All segments were then perfused intraluminally with and without l-arginine. We conducted a set of experiments with intraluminal perfusion with both l-arginine and N-nitroarginine methyl ester (L-NAME), an inhibitor of the nitric oxide – arginine pathway. Control segments were perfused extraluminally under non-ischemic conditions and intraluminally with and without l-arginine. The intraluminal perfusate contained FITC-inulin, and the fluorescence signal of FITC-inulin was measured to calculate average fluid secretion, which directly corresponds to the extent of ischemic injury. Results: Intestinal segments perfused with l-arginine had significantly decreased secretion over time in comparison to intestinal segments perfused without l-arginine (p

Summary: Background: Comprehensive characterization of exposures and immune responses to viral infections is critical to a basic understanding of human health and disease. We previously developed the VirScan system, a programmable phage-display technology for profiling antibody binding to a library of peptides designed to span the human virome. Previous VirScan analytical approaches did not carefully account for antibody cross-reactivity among sequences shared by related viruses or for the disproportionate representation of individual viruses in the library. Methods: Here we present the AntiViral Antibody Response Deconvolution Algorithm (AVARDA), a multi-module software package for analyzing VirScan datasets. AVARDA provides a probabilistic assessment of infection with species-level resolution by considering sequence alignment of all library peptides to each other and to all human viruses. We employed AVARDA to analyze VirScan data from a cohort of encephalitis patients with either known viral infections or undiagnosed etiologies. We further assessed AVARDA's utility in associating viral infection with type 1 diabetes and lupus. Findings: By comparing acute and convalescent sera, AVARDA successfully confirmed or detected encephalitis-associated responses to human herpesviruses 1, 3, 4, 5, and 6, improving the rate of diagnosing viral encephalitis in this cohort by 44%. AVARDA analyses of VirScan data from the type 1 diabetes and lupus cohorts implicated enterovirus and herpesvirus infections, respectively. Interpretation: AVARDA, in combination with VirScan and other pan-pathogen serological techniques, is likely to find broad utility in the epidemiology and diagnosis of infectious diseases. Funding: This work was made possible by support from the National Institutes of Health (NIH), the US Army Research Office, the Singapore Infectious Diseases Initiative (SIDI), the Singapore Ministry of Health's National Medical Research Council (NMRC) and the Singapore National Research Foundation (NRF).

The association between COVID-19 symptoms and antibody responses against SARS-CoV-2 is poorly characterized. We analyzed antibody levels in individuals with known SARS-CoV-2 infection to identify potential antibody-symptom associations. Convalescent plasma from 216 SARS-CoV-2 RNA+ individuals with symptomatology information were tested for the presence of IgG to the spike S1 subunit (Euroimmun ELISA), IgG to receptor binding domain (RBD, CoronaCHEK rapid test), and for IgG, IgA, and IgM to nucleocapsid (N, Bio-Rad ELISA). Logistic regression was used to estimate the odds of having a COVID-19 symptom from the antibody response, adjusting for sex and age. Cough strongly associated with antibodies against S1 (adjusted odds ratio [aOR] = 5.33; 95% CI from 1.51 to 18.86) and RBD (aOR = 4.36; CI 1.49, 12.78). In contrast, sore throat significantly associated with the absence of antibodies to S1 and N (aOR = 0.25; CI 0.08, 0.80 and aOR = 0.31; 0.11, 0.91). Similarly, lack of symptoms associated with the absence of antibodies to N and RBD (aOR = 0.16; CI 0.03, 0.97 and aOR = 0.16; CI 0.03, 1.01). Cough appeared to be correlated with a seropositive result, suggesting that SARS-CoV-2 infected individuals exhibiting lower respiratory symptoms generate a robust antibody response. Conversely, those without symptoms or limited to a sore throat while infected with SARS-CoV-2 were likely to lack a detectable antibody response. These findings strongly support the notion that severity of infection correlates with robust antibody response.

Objectives: To compare gel (Hydrasys 2 from Sebia) and capillary (Capillarys III Tera, Sebia) electrophoresis for the characterization of human serum proteins. Design and methods: 304 sera tested by gel electrophoresis during 8 routine laboratory days were concurrently tested by capillary electrophoresis. Gels were read by an experienced medical technologist while capillary profiles by a Sebia representative and the same technologist. Most sera (214 of 304, 70%) were also analyzed by immunofixation electrophoresis, used here as the gold standard to calculate sensitivity and specificity of the gel and capillary systems. Results: Gel and capillary estimated the concentration of albumin, gamma region, and M-spikes nearly perfectly, and that of beta, alpha-2, and alpha-1 regions with excellent correlation. The two systems classified concordantly 268 of 304 sera (88% agreement) as having no, one, or two M-spikes, but differed in the remaining 36 sera (12%). Gel electrophoresis correctly identified M-spikes in 82 of 112 sera that were shown to have monoclonal band(s) by immunofixation (73% sensitivity), and correctly did not reveal M-spikes in 97 of the 102 sera that had no immunofixation bands (95% specificity). Capillary achieved slightly higher sensitivity (85 of 112, 76%) and slightly lower specificity (94 of 102, 92%), but the two areas under the ROC curves were nearly identical at 0.84. Conclusions: Gel and capillary electrophoresis systems perform similarly to estimate the concentration of serum protein fractions and detect M-spikes.

Programmed cell death protein 1/programmed cell death protein ligand 1 (PD-1/PD-L1) and cytotoxic T-lymphocyte antigen 4/B7 (CTLA-4/B7) pathways are key regulators in T-cell activation and tolerance. Nivolumab, pembrolizumab (PD-1 inhibitors), atezolizumab (PD-L1 inhibitor) and ipilimumab (CTLA-4 inhibitor) are monoclonal antibodies approved for treatment of several advanced cancers. Immune checkpoint inhibitors (ICIs)-related hypophysitis is described more frequently in patients treated with anti-CTLA-4; however, recent studies reported an increasing prevalence of anti-PD-1/PD-L1-induced hypophysitis which also exhibits slightly different clinical features. We report our experience on hypophysitis induced by anti-PD-1/anti-PD-L1 treatment. We present four cases, diagnosed in the past 12 months, of hypophysitis occurring in two patients receiving anti-PD-1, in one patient receiving anti-PD-1 and anti-CTLA-4 combined therapy and in one patient receiving anti-PD-L1. In this case series, timing, clinical presentation and association with other immune-related adverse events appeared to be extremely variable; central hypoadrenalism and hyponatremia were constantly detected although sellar magnetic resonance imaging did not reveal specific signs of pituitary inflammation. These differences highlight the complexity of ICI-related hypophysitis and the existence of different mechanisms of action leading to heterogeneity of clinical presentation in patients receiving immunotherapy.

Objectives:. Current studies lack robust information on the prevalence and associated factors of cerebral microbleeds in patients who underwent extracorporeal membrane oxygenation. Design:. Retrospective analysis. Setting:. We reviewed patients who underwent (extracorporeal membrane oxygenation) and subsequent brain autopsy with gross and microscopic examinations from January 2009 to December 2018 from a single tertiary center. Patients:. Twenty-five extracorporeal membrane oxygenation patients (median age, 53 yr; interquartile range, 36–61 yr; 17 women and 8 men) underwent brain autopsy. Interventions:. Descriptive analysis of neuropathologic findings. Cerebral microbleed was defined as a small focus (< 10 mm diameter) of accumulation of blood product in the brain tissue. Macrohemorrhage was defined as any of the grossly identifiable epidural, subdural, subarachnoid, or intraparenchymal hemorrhages larger than 10 mm. Measurement and Main Results:. Of 25 (22 venoarterial extracorporeal membrane oxygenation; three venovenous extracorporeal membrane oxygenation), 15 patients (60%) were found to have cerebral microbleeds, whereas 13 (52%) had macrohemorrhages, of whom five (20%) had both. Overall, 92% of brains demonstrated the presence of either cerebral microbleeds or macrohemorrhages after extracorporeal membrane oxygenation support. Of the patients with cerebral microbleeds, lobar cerebral microbleeds (80%) occurred more frequently than deep cerebral microbleeds (60%), with 40% of patients having both types. The cases of macrohemorrhages consisted of one epidural (8%), two subdural (15%), and 10 subarachnoid hemorrhages (77%). In univariate analyses, the presence of macrohemorrhages was significantly associated with the presence of cerebral microbleeds (p = 0.03) with odds ratio of 0.13 (CI, 0.02–0.82). Age, sex, extracorporeal membrane oxygenation duration, extracorporeal membrane oxygenation type, use of aspirin or dialysis during extracorporeal membrane oxygenation support, bloodstream infections, hemoglobin, platelets, and coagulopathy profiles were not associated with cerebral microbleeds. Conclusions:. In patients with postmortem neuropathologic evaluation, 92% sustained acute cerebral microbleeds or macrohemorrhages after extracorporeal membrane oxygenation support. Cerebral microbleeds were commonly present in the majority of extracorporeal membrane oxygenation nonsurvivors. Further research is necessary to study the long-term sequelae, such as cognitive outcome of extracorporeal membrane oxygenation-associated cerebral microbleeds in extracorporeal membrane oxygenation survivors.

In transition areas, cool season turfgrasses are overseeded in autumn to maintain the high quality of dormant warm season turfgrasses, while in spring several agronomic methods (scalping, coring, topdressing, verticutting, irrigation, and targeted fertilization) or chemical desiccation are adopted to remove the cool season turfgrasses from the stand. To reduce chemical applications, several methods of “thermal weeding” have been experimented with, but little is known about these methods in zoysiagrass (Zoysia spp. Willd) spring transition. A study was conducted at the University of Pisa, Italy, on Manila grass (Zoysia matrella (L.) Merr., cv “Diamond”) (Zm) overseeded with perennial ryegrass (Lolium perenne L.) (Lp) with the aim of comparing different methods of cool season grass suppression (scalping and hot foam) and different application rates of nitrogen. To assess treatment effect, green cover, turf quality, turf color, shoot density, and some vegetation indices (GLI, DGCI and NDVI) were determined. An average green cover of at least 90% was obtained on all plots seven weeks after the treatments. While scalping had minor effects on turf appearance and on polystand composition, hot foam had a stronger effect on turf color, green cover, and turf quality in the weeks following application. Once it had recovered from the hot foam treatments, the turf had a greater number of Zm shoots and a relevant reduction of Lp shoots. The hot foam was very effective in suppressing Lp while maintaining Zm recovery capacity.

Early in the COVID-19 pandemic (March–July 2020 in Baltimore), emergency department (ED) healthcare workers (HCWs) were considered to be at greater risk of contracting SARS-CoV-2. Limited data existed, however, on the prevalence of SARS-CoV-2 infection and its impact in this workforce population. We enrolled 191 ED HCWs from a tertiary academic center, administered baseline and weekly surveys, and tested them twice (July and December 2020) for serum antibodies against SARS-CoV-2 spike protein. Approximately 6% (11 of 191, 5.8%) of ED HCWs had spike antibodies in July, a prevalence that doubled by December (21 of 174, 12.1%). A positive PCR test was self-reported by 15 of 21 (71%) seropositive and 6 of 153 (4%) seronegative HCWs (p < 0.001). Of the total 27 HCWs who had antibodies and/or were PCR positive, none required hospitalization, 18 (67%) had a self-perceived COVID-19 illness, and 12 of the 18 reported symptoms. The median number of missed workdays was 8.5 (ranging from 2 to 21). While most seropositive ED HCWs who reported symptoms took work absences, none required hospitalization, indicating that COVID-19’s impact on staffing prior to vaccination was not as great as feared.

Background: Neonatal thyrotropin (TSH) on dried blood spot (DBS), the most common screening strategy for primary congenital hypothyroidism (CH), is influenced by numerous factors that may hinder a true CH diagnosis. A second test can thus be performed to clarify the initial findings, although its application varies among screening programs.Objectives: The aim of this study was to evaluate the effect of maternal and neonatal factors on neonatal TSH levels and offer practical screening recommendations.Methods: We retrospectively analyzed screening data of 62,132 neonates born in Abruzzo, an Italian region considered mildly iodine deficient, between 2011 and 2016. We then performed a multiple linear regression to model the relationship between TSH (the dependent variable) and 13 independent variables extracted from blood collection cards.Results: Most neonates (53,551 of 62,132, 86%) had normal TSH and no clinical indications for a second screening. A minority (1,423, 2.3%) had elevated TSH in the initial DBS, which was confirmed in 97 cases (7%) on a second screen. The remaining neonates (6,594, 10.6%) had a normal initial TSH but underwent a second test in accordance with screening protocols, and were found to have delayed TSH elevation in 23 cases (0.4%). Those 120 newborns (97 + 23), considered highly suspicious for primary CH, were referred to a pediatrician for confirmatory testing and excluded from subsequent analysis of factors influencing TSH levels. Sex (β regression coefficient, β = 1.11 female to male, 95% CI 1.09, 1.12) and age at collection (β = 0.78 day 5 to days 2–3, 95% CI 0.74, 0.83) affected neonatal TSH, suggesting the utility of specific nomograms. In addition, prematurity (β = 0.85 term to preterm, 95% CI 0.80, 0.91), dopamine use (β = 0.71, 95% CI 0.62, 0.81), and birth weight (β = 1.40 normal vs. very low, 95% CI 1.05, 1.89) strongly influenced neonatal TSH.Conclusions: Neonatal TSH is influenced by several factors supporting the delineation of local sex- and age-adjusted TSH cutoffs, and the universal adoption of a second TSH test in neonates at risk of missed primary CH diagnosis.

Abstract Background Malignant extrarenal rhabdoid tumor of the gastrointestinal tract is rarely reported in the literature. It is characterized by poor prognosis and aggressive metastatic features. A literature review evidenced only 19 cases, with poor outcome. Case presentation We report a case of a colonic “pure” malignant extrarenal rhabdoid tumor with metastatic nodes in a 65-year-old Caucasian man. He was treated surgically with no recurrence, no adjuvant chemotherapy, and with 4-year survival without disease at the time of the submission of this article. Conclusions We present an extraordinary case of long-term survival due to the extended surgical treatment. We believe that the absence of organ metastasis at presentation is a positive prognostic factor, although pathology confirmed node involvement (13/38 positive) on microscopy.

Tall fescue (Schedonorus arundinaceus (Schreb.) Dumort.) is often managed with a cutting height ranging from 70 to 100 mm in ornamental lawns. Some autonomous mowers have been specifically designed to maintain mowing height in the same range. Generally, autonomous mowers operate by following random trajectories, and substantial overlapping is needed to obtain full coverage of the working area. In the case of tall grass, this may cause lodging of grass plants, which in turn may reduce turf quality. The introduction of a navigation system based on systematic trajectories has the potential to improve the performances of autonomous mowers with respect to machine efficiency and turf quality. With the aim of determining the effects of reduced mowing frequency and systematic navigation systems on turf quality and mower performances in terms of working time, energy consumption and overlapping, the performances of two autonomous mowers working with random and systematic trajectories were tested on a mature tall fescue lawn at 90 mm cutting height. The working efficiency was approximately 80% for the systematic trajectories and approximately 35% for the random trajectories; this was mainly due to the lower overlapping associated with systematic trajectories. Turf quality was slightly higher for the mower working systematically (a score of 8 using a 1–9 score with 1 = poor, 6 = acceptable and 9 = best) compared to the one working randomly (quality of 7 and 6 on a 1–9 scale with 1 = poor and 9 = best). No appreciable lodging was observed in either case. For tall, managed lawns, systematic trajectories may improve autonomous mowers’ overall performances.

Purpose: Diagnostic criteria for monoclonal gammopathy of undetermined significance (MGUS) do not currently include ocular phenotypic changes. Here, we offer a new diagnostic approach that is useful in patients with posteriorly located corneal depositions and present evidence to support the theory that the aqueous humor is a source for monoclonal proteins accumulated in the cornea. Observations: A 77-year-old woman presented to the clinic with a gradual decrease in visual acuity over 6 months. Slit lamp examination revealed bilateral central guttae consistent with Fuchs corneal dystrophy, peripheral circular band-like corneal opacities in the deep stroma, and bilateral nuclear sclerotic and cortical cataracts. Anterior segment optical coherence tomography confirmed corneal opacities in the posterior stroma and Descemet membrane. Immunological studies revealed increased serum IgG levels of 3220 mg/dL and serum electrophoresis showed an abnormal monoclonal band of 2.4 g/dL identified as IgG lambda by immunofixation electrophoresis. The patient was referred to the hematology clinic where she underwent further systemic workup and was diagnosed with MGUS. Immunofixation electrophoresis of aqueous sampling, which was performed at the time of cataract surgery, confirmed the presence of the IgG lambda gammopathy in the anterior chamber. Conclusions and importance: Monoclonal gammopathy, although rare, should be included in the differential diagnosis of corneal opacities, as the ocular finding can be the initial manifestation of a systemic disease that can potentially be life-threatening. When corneal biopsy is not feasible due to the location of corneal pathology, aqueous sampling may be an alternative approach towards a clinical diagnosis. We propose a new terminology, “monoclonal gammopathy of ocular significance,” for patients diagnosed with MGUS, however, their only significant clinical finding is ocular manifestation. Keywords: Cornea, Deposition, Monoclonal gammopathy of undetermined significance, Paraproteinemic keratopathy, Lambda, Immunofixation

Weed control is crucial to ensure that turfgrass is established effectively. Although herbicides are commonly used to control weeds in turfgrasses, environmental and public health concerns have led to limiting or banning the use of synthetic herbicides in urban areas. The species seashore paspalum (Paspalum vaginatumSw.) is susceptible to such herbicides. Flame weeding could be an alternative to the use of synthetic herbicides for selective weed control in seashore paspalum. In this study, five different liquefied petroleum gas (LPG) doses of flaming (0, 61, 91, 157 and 237 kg ha-1) were tested in order to find the optimal dose, in terms of weed control and costs. The aim was to maintain a seashore paspalum (cultivar ‘Salam’) turf free of weeds during spring green-up, and at the same time avoid damaging the turfgrass. Using a self-propelled machine designed and built at the University of Pisa, flaming was applied three times when weeds started growing and the turfgrass started green-up. Our results highlight that an LPG dose of 157 kg ha-1was the most economic dose that led to a significant reduction in initial weed cover and density, enabling the turfgrass to recover three weeks after the third application.

BackgroundThe calcium-sensing receptor (CaSR) has been localized and characterized in numerous tissues throughout the body. In the mammalian gastrointestinal tract, the CaSR is known to act as a nutrient sensor and has recently been found to play a role in intestinal fluid and electrolyte balance. This study aims to demonstrate the functionality of the CaSR as a modulator of fluid secretion and absorption along the small intestine.MethodsSmall intestine regions (proximal, middle, and distal) were isolated from Sprague Dawley rats and loaded into an ex vivo intestinal perfusion device that provides independent intraluminal and extraluminal (serosa/basolateral) perfusion. The regions were perfused with 5 and 7 mM of Ca2+, both in the presence and absence of forskolin (FSK), a potent secretagogue. Control experiments were conducted with intraluminal perfusate containing standard Ringer-HEPES buffer with a physiological concentration of Ca2+ (1 mM). A second set of comparison experiments was performed with intraluminal perfusates containing AC-265347, a CaSR activator and agonist, in the presence of FSK. In all experimental conditions, the intraluminal perfusate contained fluorescein isothiocyanate (FITC)-inulin, a nonabsorbable fluorescent marker of secretion and/or absorption. Intraluminal fluorescence signal was utilized as a measure of water movement at the start of the experiment and every 15 min for 90 min.ResultsUnder physiological conditions, increasing the concentration of Ca2+ in the luminal perfusate reduced intestinal fluid secretion in all regions. At a Ca2+ concentration of 7 mM, net fluid absorption was observed in all regions. In the presence of FSK, 5 mM Ca2+ significantly decreased fluid secretion and 7 mM Ca2+ abolished FSK-induced fluid secretion. Intraluminal perfusion with 5 mM Ca2+ was as effective as AC-265347, in reducing secretagogue-induced fluid hypersecretion in the proximal and middle regions.ConclusionThis study concludes that apical CaSR is active along the small intestine. Its activation by Ca2+ and/or calcimimetics reduces fluid secretion in a dose-dependent manner, with higher Ca2+ concentrations, or application of a calcimimetic, leading to fluid absorption. We furthermore show that, in the presence of FSK, receptor activation abates FSK secretagogue-induced fluid secretion. This presents a new therapeutic target to address secretory diarrheal illnesses.

Warm season turfgrasses can be grown successfully in the transition zone, but dormancy occurs to some extent during the winter. Overseeding with cool-season turfgrasses is necessary if winter dormancy of warm season turfgrasses is not tolerated. The increasing availability of zoysiagrass cultivars has enabled this genus to be considered suitable for low-maintenance golf courses, especially for golf tees and golf fairways. On the other hand, zoysiagrasses have the most rigid leaves of all turfgrass species, followed by bermudagrasses and by the other warm season turf species. Thus, to have a high mowing quality, mowers working constantly on zoysiagrasses require more sharpening than mowers working on other grasses. Rotary mowers are not suitable for mowing at low heights and often result in scalping, while reel mowers perform optimal mowing at a short height (below 2.5 cm) but require accurate management and frequent sharpening. Autonomous mowers have proven to produce a superior turf quality compared with traditional walk-behind rotary mowers, but no autonomous mower has ever been tested at a low mowing height on an overseeded warm season turfgrass. Because of this, the trial was carried out to simulate a golf tee overseeded with cool season turfgrasses, with low input fertilisation rates and with one of the most difficult turf species to mow; i.e. Zoysia matrella (L.) Merr. The trial was carried out in San Piero a Grado (Pisa, Italy) from October 2016 to October 2018. After a two-year period the best turf quality was achieved with Festuca rubra L. ssp. cultivars among the overseeded species, especially during fall. In many cases turf quality increased after manila grass green up since the combination of both cool season and warm season species gave a higher quality to the turfgrass, due to the finer leaf texture and higher shoot density of some cool season species. Moreover, recovery of manila grass ground cover was satisficing. In conclusion, a polystand of manila grass and Festuca rubra ssp. could be suitable for golf tees with low-input management.

The use of warm season turfgrasses is a consolidated trend in the climatic transition zone of Mediterranean countries, in particular St. Augustinegrass (Stenotaphrum secundatum (Walt.) Kuntze) begins to be widespread in warm coastal areas. However, little is known about the performance of the different cultivars of this species in southern Europe. In 2016-2017 a trial was carried out in three locations in Italy, Padova, Pisa, and Palermo, located in the north, center and south of the country respectively. Four cultivars (Floratine, Captiva, Sapphire, Palmetto) and five ecotypes (CeRTES 201, CeRTES 202, CeRTES 203, CeRTES 204, CeRTES 205) were compared in terms of their growth characteristics and morphological traits during establishment. The results highlighted that stolon growth was significantly affected by the location, as well as green colour retention. Stolon growth rate, internode length and internode volume and turf quality were, however, significantly determined by the accession effect. The quality of the ecotypes was also in some cases comparable to that of the cultivars. In Padova, winterkill occurred in most of the accessions, while in Pisa and Palermo, all the entries survived. In conclusion, St. Augustinegrass is suitable for turf use in the central and southern coastal area of Italy.

Enhancing vineyards sustainability and reducing herbicides usage is a crucial theme, thus alternative weed management methods are starting to be studied. Cover crops have been shown to provide for several environmental services such as performing an efficient weed control and promoting biodiversity, thus improving the sustainability of the overall management system. However, the use of cover crops is usually confined to the interrow area in order to avoid competition with vines. Under-trellis weed management in vineyards is an important challenge, conventionally fulfilled by cultivation or repeated herbicides applications. Autonomous mowers are small autonomous machines that have shown a great efficiency when employed in agricultural contexts. Due to their reduced size, they can easily prevent the excessive vertical growth of weeds both in the interrows and under trellis without the use of chemical applications. The aim of this trial was to evaluate if the combination of cover crop species and autonomous mowers management could improve vineyard sustainability. Vineyard floor cover that was managed with an autonomous mower had a shorter canopy height and a lower weed dry biomass compared to the conventionally managed vineyard floor cover. Vineyard floor management providing cover crops and autonomous mowing had a significantly lower weed cover percentage compared to conventional floor management, especially under-trellis. The results of this trial suggest that the combination of cover crops and autonomous mowers may be a sustainable and reliable technique to include in vineyards floor management.

The establishment of permanent cover crops is becoming a common practice in vineyard floor management. Turfgrass science may provide species and techniques with a high potential for improving the sustainability of vineyard floor management. Based on this assumption, an experiment was carried out during 2018 and 2019 at the Donna Olimpia Vineyard, Bolgheri, Italy. The trial aimed at comparing an innovative floor management system based on a turf-type cultivar of bermudagrass mown with an autonomous mower with a conventional floor management system. Ground cover percentage, energy consumption, CO2 emissions, grapevine water status, leaf nitrogen content, fruit yield and must composition have been assessed in order to perform the comparison. The innovative vineyard floor management produced an almost complete ground cover (98%) at the end of the second growing season, with the resident species reduced to a small percentage (4%). Resident species growing under-trellis were efficiently controlled without herbicide applications. A lower primary energy consumption and a reduction in CO2 emissions were observed for the innovative management system compared to the conventional management system. Grapevine water status, leaf chlorophyll content, soil–plant analyses development (SPAD), fruit yields and must composition were similar between the different soil management systems. Based on results obtained in this trial, turf-type bermudagrass and innovative mowing machines may contribute to enhance the sustainability of vineyard floor management.

Autonomous mowers are becoming increasingly common in public and private greenspaces. Autonomous mowers can provide several advantages since these machines help to save time and energy and prevent operators from possible injuries. Current autonomous mowers operate by following random trajectories within areas defined by a shallow-buried boundary wire that has the purpose to generate an electro-magnetic field. Once the electro-magnetic field is perceived by the autonomous mower, the machine will stop and change direction. Mowing along random trajectories is considered an efficient solution to manage areas with a variable number of obstacles. In agriculture, autonomous technologies are becoming increasingly popular since they can help to increase both the quantity and quality of agricultural products by reducing productive cost and improving the production process. Thus, even autonomous mowers may be useful to carry out some of the agricultural operations that are highly time consuming. In fact, some autonomous mowers designed and realized to work in vineyards and home vegetable gardens are already available on the market. The aim of this study was to compare the work capacity of six autonomous mowers that move along random trajectories in areas with a high number of obstacles to assess if these machines may be employed in some agricultural contexts. The six autonomous mowers were split in three groups based on their size (large, medium, and small) and were left to work in two areas with equal number of obstacles but different layouts. The first area (Site A) had a square shape and an extension of 23.04 m2, in order to keep the autonomous mowers enclosed inside it. The second area (Site B) had a square shape and an extension of 84.64 m2, in order to have a part of the area with no obstacles. The layout and the size of the two areas affected the autonomous mowers performances in different ways. The six autonomous mowers working on Site A obtained similar results and higher performances compared to the same mowers working on Site B. All the autonomous mowers proved to be able to mow more than 89% of Site A after 2 h and more than 98% of Site A after 5 h. On Site B small size autonomous mowers obtained the best results mowing more than 83% of the area with obstacles after 2 h and more than 98% of the area with obstacles after 5 h. However, specific work settings allowed larger autonomous mowers to improve their efficiency, obtaining similar results compared to smaller autonomous mowers.

Pancreatic intraductal papillary-mucinous neoplasm is a rare primary neoplasm of unknown pathogenesis. This kind of tumor represents 0.2–2.7% of all pancreatic cancers and they may proceed to malignant lesions. In this study, we describe a case of pancreatic intraductal papillary-mucinous tumor (4.3 cm) with normal tumoral markers and nuclear atypia. We perform also a systematic review of the literature on MEDLINE and find only one relevant study that used microwave ablation for the palliative treatment of pancreatic tumor. We describe the case of a 70-year-old Caucasian male who was diagnosed with a pancreatic tumor with biliary tree dilatation. The patient underwent computed tomography (CT), percutaneous biopsy, and an endoscopic positioning of prosthesis in the biliary tree. Due to the worsening of jaundice and cholestasis, and considering the severe systemic disease status, palliative surgery with microwave thermoablation in the head of pancreas was performed. No complications were observed. The hospitalization lasted for 11 days after surgery, with normal liver and pancreatic lab tests at discharge. The patient followed a line of chemotherapy for 6 months with a complete response for 8 months. One month after the treatment, a staging CT scan was performed showing the size of the cephalopancreatic lesion had decreased from 43 to 35 mm with signs of complete ablation. The patient had a total response at the imaging of 10 months. One year later, a CT scan follow-up showed progression of the pancreatic disease. The disease remained stable for 18 months. The patient died due to cardiovascular complications with an overall survival of 30 months. Microwave ablation in our case report has been demonstrated to be feasible and safe without complications. It can be used as a phase of multimodality treatment in patients with severe systemic disease status and advanced intraductal papillary-mucinous neoplasm.