[18F]-FDG PET/CT in oncologic patients with unsuspected asymptomatic infection with SARS-CoV-2.

PURPOSE: Spain has been one of the most affected countries by the COVID-19 pandemic, being among the countries with worse numbers, including the death rate. However, most patients are asymptomatic, although they are very contagious. The objective of this study was to investigate the incidence in oncological patients infected with SARS-CoV-2 that are asymptomatic for COVID-19 and at home and that undergo PET/CT for oncologic indications, nonrelated to COVID-19, finding in the PET/CT lung alterations that are suggestive of SARS-CoV-2 infection. METHODS: During the period of maximum incidence of the global pandemic in one of the most affected regions of Spain, there were 145 patients that met inclusion and exclusion criteria and were included in the study. Imaging findings previously described such as ground-glass opacities with low [18F]-FDG uptake were considered images suspicious for SARS-CoV-2 infection. Patients with these findings were referred to RT-PCR testing and close follow-up to confirm the presence or absence of COVID-19. RESULTS: Suspicious lung imaging findings were present in 7 of 145 patients (4.8%). Five of these 7 patients were confirmed as presenting SARS-CoV-2 infection, this is, COVID-19. In the remaining two, it was not possible to confirm the presence of COVID-19 with RT-PCR, although in one of them, PET/CT allowed an early diagnosis of a lung infection related to a bacterial pneumonic infection that was promptly and adequately treated with antibiotics. CONCLUSION: These results confirm that the prevalence of SARS-CoV-2 infection is higher than suspected and that there are asymptomatic patients that are attending imaging departments to be explored for their baseline oncologic processes. In these patients, PET/CT allows an early diagnosis of COVID-19.

Effects of cannabis tetrahydrocannabinol on endocannabinoid homeostasis in human placenta.

Cannabis use has become a hot topic in several countries due to the debate about its legalization for medical purposes. However, data are limited regarding adverse events, safety and potential impact on reproductive health. Cannabis consumption during pregnancy has been associated with gestational disorders such as preterm birth, intrauterine growth restriction, low birth weight and increased risk of miscarriage, though the underlying biochemical mechanisms are still unknown. Given that the endocannabinoid system (ECS) is involved in several reproductive processes, we tested the hypothesis that the negative outcomes may result from the impact on the ECS homeostasis caused by the main psychoactive compound of cannabis, Δ9-tetrahydrocannabinol (THC). We demonstrate that THC (10-40 µM) impairs placental endocannabinoid system by disrupting the endocannabinoid anandamide (AEA) levels and the expression of AEA synthetic and degrading enzymes N-arachidonoylphosphatidylethanolamine-specific phospholipase D (NAPE-PLD) and fatty acid amide hydrolase (FAAH), respectively. Although, no alterations in cannabinoid receptors CB1 and CB2 expression were observed. Thus, long-term local AEA levels are associated with a shift in the enzymatic profile to re-establish ECS homeostasis. In chronic cannabis users, high AEA levels in placenta may disturb the delicate balance of trophoblast cells turnover leading to alterations in normal placental development and foetal growth.

Anandamide oxidative metabolism-induced endoplasmic reticulum stress and apoptosis.

The Endocannabinoid System (ECS) has been recognized as a crucial player in human reproduction. Changes in the levels of anandamide (AEA), the main endocannabinoid (eCB), negatively affect reproductive events, such as implantation, decidualization and placentation. Cyclooxygenase-2 (COX-2) is a major enzyme expressed in the endometrium and its involvement in female reproductive system has evolved over the last few years. Currently, COX-2 oxidative metabolism is emerging as a key mediator of AEA-induced actions. In this study, we aimed to disclose the mechanisms underlying the effects of AEA in human endometrial stromal cell fate, using a human-derived endometrial cell line (St-T1b). We found that AEA has an anti-proliferative activity through a direct effect on cell cycle progression by inducing G2/M arrest. Moreover, high levels of AEA increased COX-2 activity, triggering apoptotic cell death, with loss of mitochondrial membrane potential, induction of caspase -9 and -3/-7 activities, and cleavage of poly (ADP-ribose) polymerase (PARP). In addition, the involvement of intracellular reactive oxygen species (ROS) and endoplasmic reticulum (ER) stress was verified. These effects were prevented by pre-incubation with a selective COX-2 inhibitor. Therefore, we hypothesize that, in response to altered levels of this eCB, COX-2 oxidative metabolism of AEA may deregulate endometrial cell turnover and, consequently, interfere with cellular events crucial for implantation and decidualization, with a negative impact on human fertility.

Anandamide and decidual remodelling: COX-2 oxidative metabolism as a key regulator.

Recently, endocannabinoids have emerged as signalling mediators in reproduction. It is widely accepted that anandamide (AEA) levels must be tightly regulated, and that a disturbance in AEA levels may impact decidual stability and regression. We have previously characterized the endocannabinoid machinery in rat decidual tissue and reported the pro-apoptotic action of AEA on rat decidual cells. Cyclooxygenase-2 (COX-2) is an inducible enzyme that plays a crucial role in early pregnancy, and is also a key modulator in the crosstalk between endocannabinoids and prostaglandins. On the other hand, AEA-oxidative metabolism by COX-2 is not merely a mean to inactivate its action, but it yields the formation of a new class of mediators, named prostaglandin-ethanolamides, or prostamides. In this study we found that AEA-induced apoptosis in decidual cells involves COX-2 metabolic pathway. AEA induced COX-2 expression through p38 MAPK, resulting in the formation of prostamide E2 (PME2). Our findings also suggest that AEA-induced effect is associated with NF-kB activation. Finally, we describe the involvement of PME2 in the induction of the intrinsic apoptotic pathway in rat decidual cells. Altogether, our findings highlight the role of COX-2 as a gatekeeper in the uterine environment and clarify the impact of the deregulation of AEA levels on the decidual remodelling process.

The endocannabinoid anandamide impairs in vitro decidualization of human cells.

Endocannabinoids (eCBs) are endogenous mediators that along with the cannabinoid receptors (CB1 and CB2), a membrane transporter and metabolic enzymes form the endocannabinoid system (ECS). Several eCBs have been discovered with emphasis on anandamide (AEA). They are involved in several biological processes such as energy balance, immune response and reproduction. Decidualization occurs during the secretory phase of human menstrual cycle, which involves proliferation and differentiation of endometrial stromal cells into decidual cells and is crucial for the establishment and progression of pregnancy. In this study, a telomerase-immortalized human endometrial stromal cell line (St-T1b) and non-differentiated primary cultures of human decidual fibroblasts from term placenta were used to characterize the ECS using immunoblotting and qRT-PCR techniques. It was shown that St-T1b cells express CB1, but not CB2, and that both receptors are expressed in HdF cells. Furthermore, the expression of fatty acid amide hydrolase (FAAH), the main degrading enzyme of AEA, increased during stromal cell differentiation. AEA inhibited cell proliferation, through deregulation of cell cycle progression and induced polyploidy. Moreover, through CB1 binding receptor, AEA also impaired cell differentiation. Therefore, AEA is proposed as a modulator of human decidualization. Our findings may provide wider implications, as deregulated levels of AEA, due to Cannabis sativa consumption or altered expression of the metabolic enzymes, may negatively regulate human endometrial stromal cell decidualization with an impact on human (in)fertility.Free Portuguese abstract: A Portuguese translation of this abstract is freely available at http://www.reproduction-online.org/content/152/4/351/suppl/DC1.

Rat spontaneous foetal resorption: altered α2-macroglobulin levels and uNK cell number.

During rat pregnancy, some of the foetoplacental units undergo complete spontaneous resorption while the adjacent units remain unaffected. In an attempt to clarify the mechanisms implicated in this spontaneous resorption, implantation units from days 14 and 16 of pregnancy were examined. The number of implantation sites and resorption units was recorded, and uterine paraffin sections were stained with haematoxylin and eosin for the evaluation of tissue morphology. The incidence of resorption was about 9.2 % on day 14 and 8.2 % on day 16. Perforin and active caspase-3 immunostaining were performed for localization and characterization of uterine natural killer (uNK) and apoptotic cells, respectively. The α2-macroglobulin (α2-MG) expression was examined by in situ hybridization, immunohistochemistry and its levels quantified by enzyme-linked immunosorbent assay. A reduction in α2-MG decidual levels in resorpted units was observed when compared to normal implantation units in both days. This potent protease inhibitor is the major product secreted by the mesometrial decidual tissue and may constitute an indicator of maternal tissues remodelling abnormalities. Besides the decreased α2-MG levels, an increase in uNK cell number was found in resorption units. The decreased α2-MG levels may be related to the aberrant control of trophoblast invasion that may activate uNK cells. The elucidation of the mechanisms underlying natural pregnancy loss in rat may contribute for the clarification of the "vanishing twin" phenomenon that occurs in human pregnancy.

Endogenous cannabinoids revisited: a biochemistry perspective.

Marijuana is the most commonly used illegal drug, particularly in Western societies. The discovery of an endogenous cannabinoid system (ECS) highlighted new molecules in various physiological processes. The ECS consists of G-protein-coupled cannabinoid receptors that can be activated by small lipid mediators, termed endocannabinoids (eCBs) and cannabis-derived drugs, plus the associated biochemical machinery (precursors, synthesis and degradative enzymes, and transporters). Several biochemical, pharmacological and physiological studies have shown that endocannabinoid system elements are widely distributed throughout the body, with regional variations and organ-specific actions. This review portrays the endocannabinoid "family" on new studies concerning eCB storage, release and functional roles and on the growing importance of its bioactive metabolites. Those findings reinforce and confirm the importance of ECS. Strategies for manipulating the system for the treatment of human disease will require a thorough understanding of the roles of the different eCBs and their sources.

Prevalence of Falls and Associated Factors among Community-Dwelling Older Adults: A Cross-Sectional Study.

AIM: As a person ages, the risk of falls increases, which affects quality of life and represents a financial burden to health- and social-systems, and a greater morbidity and mortality risk. Falls lead to decreased social contact, anxiety, long-term physical disability, severe dependency and hospitalizations. Currently, few studies address this phenomenon using a uniform methodology; therefore, this study aims to explore the prevalence of falls and associated-variables in older adults across Europe. METHODS: In this cross-sectional analysis, we used data from Wave 6 of SHARE. The prevalence of falls was assessed through the answer "falling down" to the question "For the past six months at least, have you been bothered by any of the health conditions on this card?". Multilevel logistic regression was used, using falls as a dependent variable. Multilevel univariable logistic regression models were made to identify potential associated factors. RESULTS: From the 41,098 participants, 56.3% were female, and the average age was of 70.0 ± 8.9 years. The prevalence of falls was 8.2% (CI 8.0% to 8.4%), being higher in women (10.1% vs. 5.8%) and increasing with age. Age, female gender, being frail or pre-frail, higher scores on the EURO-D scale, polypharmacy and fear of falling were found to be significantly associated with falls. CONCLUSIONS: We found that falls are prevalent in the European community-dwelling population, with variations between countries. As a public health priority, identification of the variables associated with falls is important in order to identify/monitor the risk in older groups and develop tailored and cost-effective interventions for falls prevention.

Gas chromatography-mass spectrometry analysis of nine bisphenols in canned meat products and human risk estimation.

The presence of bisphenol A (BPA) and eight BPA analogues was investigated in 30 canned meat samples (sausages, pâtés, and whole meals). For that, a previously developed gas chromatography-mass spectrometry (GC-MS) methodology based on a quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction followed by dispersive liquid-liquid microextraction (DLLME) with "in situ" acetylation was optimized, namely by the introduction of a novel solid-phase dispersive sorbent. Results showed that all the samples were contaminated with at least one compound from the five bisphenol analogues found (BPA, BPB, BPF, BPAF, and BPZ). Nineteen samples showed the simultaneous presence of BPA and one or more analogues, with a maximum of four different compounds in two of the samples. In half of the samples, the sum of all bisphenols was higher than 50 µg/kg, with a maximum of 236 µg/kg. Regarding human food safety, the estimated daily intake (EDI), target hazard quotient (THQ) and hazard index (HI) assessed were higher than those established by the European Food Safety Authority suggesting hazard risk for human consumers.

18F-FDG PET/CT in Hodgkin Lymphoma With Unsuspected COVID-19.

We present an asymptomatic 70-year-old man referred for an F-FDG PET/CT for initial staging of a Hodgkin lymphoma. F-FDG PET/CT showed bilateral cervical lymphadenopathy (stage II). Incidentally, the CT demonstrated bilateral ground-glass opacities with low-grade F-FDG activity. CT findings were suspicious for COVID-19 pneumonitis. The COVID-19 reverse transcriptase polymerase chain reaction (RT-PCR) examination result was negative. Given the high clinical suspicion for COVID-19, the patient was isolated and repeat RT-PCR was positive at 72 hours. RT-PCR may be falsely negative in early COVID-19 disease, even with positive CT findings.

Peptide functionalized magneto-plasmonic nanoparticles obtained by microfluidics for inhibition of ß-amyloid aggregation.

In the present work, we report on the synthesis of peptide functionalized magneto-plasmonic nanoparticles in a simple microfluidic platform. Superparamagnetic nanoparticles and gold nanorods were selected for this study. Magnetic nanoparticles were functionalized with peptide D1, which can bind selectively to toxic aggregates of the ß-amyloid peptide associated with Alzheimer's disease. Gold nanorods were functionalized with chitosan replacing the surfactant cetyltrimethylammonium bromide to reduce the cytotoxic effect. The selected microfluidic strategy yields structures with plasmonic and magnetic properties in a nanostructure. Cytotoxic assays with SH-SY5Y cells demonstrate that nanoparticles obtained by microfluidics do not affect cell viability at the studied concentrations. Additionally, these magneto-plasmonic nanoparticles inhibit fibril formation demonstrating that the magneto-plasmonic nanoparticles obtained by microfluidics could be applied for a potential treatment and diagnosis of Alzheimer's disease.

The endocannabinoid system expression in the female reproductive tract is modulated by estrogen.

The endocannabinoid system (ECS) is involved in several physiological events that resulted in a growing interest in its modulation. Moreover, the uterine levels of anandamide (AEA), the major endocannabinoid, must be tightly regulated to create proper embryo implantation conditions. However, there are no evidences about the regulation of AEA in uterus by estrogen. Thus, the aim of this study is to elucidate whether estradiol benzoate (EB) and tamoxifen (TAM) administration to ovariectomized (OVX) rats can induce changes in the expression of cannabinoid receptors and AEA-metabolic enzymes in uterus by evaluating gene transcription and protein levels by qPCR, Western blot and immunohistochemistry. Moreover, the plasmatic and uterine levels of AEA and of prostaglandin E2 (PGE2) and prostaglandin F2α (PGF2α), the major cyclooxygenase-2 (COX-2) products, were determined by UPLC-MS/MS. The immunohistochemistry showed that cannabinoid receptors, as well as AEA-metabolic enzymes are mainly located in the epithelial cells of both lumen and glands and, to a lesser extent, in the muscle cells. Moreover, EB administration to OVX rats significantly increased CB1, CB2, NAPE-PLD, FAAH and COX-2 expression and transcription. These effects were absent in TAM and TAM+EB treatments showing that this response is estrogen receptor dependent. Additionally, although uterine levels of AEA remained unchanged in EB or TAM treated animals, they showed a rise with EB treatment in plasma. The latter also produced a decrease in uterine PGE2 levels. In summary, these data collectively indicate that the expression of ECS components, as well as, the AEA and PGE2 levels in rat uterus is modulated by EB. Thus, estradiol may have a direct regulatory role in the modulation of ECS in female reproductive tissues.

Photothermal conversion efficiency and cytotoxic effect of gold nanorods stabilized with chitosan, alginate and poly(vinyl alcohol).

Gold nanorods (GNR) use has been proposed in medical applications because of their intrinsic photothermal properties. However, the presence of CTAB molecules adsorbed onto the surface of GNRs results in a highly cytotoxic GNR system. In this work we replace the CTAB molecules with a thiolated chitosan. Once chitosan coated GNRs (Chi-SH-GNR) were attained, a film of alginate (Alg-Chi-SH-GNR) or polyvinyl alcohol (PVA-Chi-SH-GNR) was deposited onto the surface of Chi-GNR by a layer-by-layer process. The photothermal conversion efficiency for the GNR systems was determined irradiating the GNRs suspended in aqua media with a CW 808nm diode laser (CNI, China). The cytotoxicity effect and the photothermal cellular damage of GNR systems were evaluated on a breast cancer cell line. Results show that polymer coats did not affect the transduction photothermal efficiency. Values around 50% were obtained for the different coated gold nanorods. The cytotoxicity of coated gold nanorods diminished significantly compared with those GNR stabilized with CTAB. The laser irradiation of cells treated with gold nanorods showed a decrease in their viability compared with the cells treated but no irradiated.

Inflammatory pseudotumour of the infratemporal fossa visualized with 18F-FDG PET/CT.

The inflammatory pseudotumour of the head and neck is a benign lesion, extremely rare outside the cranial orbits. A case is presented of an inflammatory pseudotumour not associated with the IgG4-related disease. The pseudotumour was found as a solitary mass in the infratemporal fossa of a young woman who complained of otalgia and hearing loss. A clear image of the lesion was obtained using an 18F-fluoro-deoxy-glucose (18F-FDG) PET. After the histopathological diagnosis, and treatment with corticosteroids, a second 18F-FDG PET was performed. The metabolic image had returned to normal, and the previously observed mass disappeared. A brief review is presented of the studies examining this type of lesion.

Clinical and genetic linkage analysis of a large Venezuelan kindred with Usher syndrome.

OBJECTIVE: To undertake a comprehensive investigation into the very high incidence of congenital deafness on the Macano peninsula of Margarita Island, Venezuela. METHODS: Numerous visits were made to the isolated island community over a 4-year-period. During these visits, it became apparent that a significant number of individuals complained of problems with hearing and vision. Socioeconomic assessments, family pedigrees and clinical histories were recorded on standard questionnaires. All individuals underwent thorough otolaryngologic and ophthalmologic examinations. Twenty milliliters of peripheral venous blood was obtained from each participant. A genome-wide linkage analysis study was performed. Polymorphic microsatellite markers were amplified by polymerase chain reaction and separated on polyacrylamide gels. An ABI 377XL sequencer was used to separate fragments and LOD scores were calculated by using published software. RESULTS: Twenty-four families were identified, comprising 329 individuals, age range 1-80 years, including 184 children. All families were categorized in the lower two (least affluent) socioeconomic categories. A high incidence of consanguinity was detected. Fifteen individuals (11 adults, 4 children) had profound congenital sensorineural hearing loss, vestibular areflexia and retinitis pigmentosa. A maximum LOD score of 6.76 (Linkage >3.0), between markers D11s4186 and D11s911, confirmed linkage to chromosome 11q13.5. The gene myosin VIIA (MYO7A) was confirmed in the interval. Clinical and genetic findings are consistent with a diagnosis of Usher syndrome 1B for those with hearing and vision problems. CONCLUSIONS: We report 15 Usher syndrome 1B individuals from a newly detected Latin American socio-demographic origin, with a very high prevalence of 76 per 100,000 population.

The Endocannabinoid System in the Postimplantation Period: A Role during Decidualization and Placentation.

Although the detrimental effects of cannabis consumption during gestation are known for years, the vast majority of studies established a link between cannabis consumption and foetal development. The complex maternal-foetal interrelationships within the placental bed are essential for normal pregnancy, and decidua definitively contributes to the success of this process. Nevertheless, the molecular signalling network that coordinates strategies for successful decidualization and placentation are not well understood. The discovery of the endocannabinoid system highlighted new signalling mediators in various physiological processes, including reproduction. It is known that endocannabinoids present regulatory functions during blastocyst development, oviductal transport, and implantation. In addition, all the endocannabinoid machinery was found to be expressed in decidual and placental tissues. Additionally, endocannabinoid's plasmatic levels were found to fluctuate during normal gestation and to induce decidual cell death and disturb normal placental development. Moreover, aberrant endocannabinoid signalling during the period of placental development has been associated with pregnancy disorders. It indicates the existence of a possible regulatory role for these molecules during decidualization and placentation processes, which are known to be particularly vulnerable. In this review, the influence of the endocannabinoid system in these critical processes is explored and discussed.

Modulation of the novel cannabinoid receptor - GPR55 - during rat fetoplacental development.

Decidualization process involves the morphological and functional transformation of endometrial stromal cells into decidual cells. This is a finely regulated process, which involves proliferation and differentiation of stromal cells into decidual cells, which is followed by regression of the decidual tissue, mainly by apoptosis, necessary to accommodate the growing embryo. Together with the endogenous cannabinoids (ECs) and the respective metabolizing-enzymes, the cannabinoid receptors complete the endocannabinoid system (ECS). Two cannabinoid receptors have been described so far, CB1 and CB2, though a third has been suggested, CB3. Although the ECS role in several biological functions, including reproductive processes, is now well documented, the current state of knowledge about this system is still incomplete. In order to investigate the expression of GPR55, referred as the novel cannabinoid receptor 3 (CB3), in the uterine maternal tissues during normal pregnancy we analysed its expression by Q-PCR, Western blot and immunohistochemistry during fetoplacental period. We found higher protein levels on day 14, after full development of mesometrial decidua. In addition, GPR55 was found in uterine natural killer (uNK) cells pointing to an involvement in the immunological reactions that occur during pregnancy. The prominent expression of GPR55 in decidual cells suggests a role in mediating cannabinoid signalling during fetoplacental development. Additionally, we have studied the effects resulting from its activation in primary decidual cell cultures, which revealed a potential modulation of cell viability through GPR55. The data presented here may clarify the role of GPR55 in fetoplacental development and highlights the presence of a new target for cannabinoid signalling during pregnancy.