Inhibition of LTßR signalling activates WNT-induced regeneration in lung.

Lymphotoxin ß-receptor (LTßR) signalling promotes lymphoid neogenesis and the development of tertiary lymphoid structures1,2, which are associated with severe chronic inflammatory diseases that span several organ systems3-6. How LTßR signalling drives chronic tissue damage particularly in the lung, the mechanism(s) that regulate this process, and whether LTßR blockade might be of therapeutic value have remained unclear. Here we demonstrate increased expression of LTßR ligands in adaptive and innate immune cells, enhanced non-canonical NF-κB signalling, and enriched LTßR target gene expression in lung epithelial cells from patients with smoking-associated chronic obstructive pulmonary disease (COPD) and from mice chronically exposed to cigarette smoke. Therapeutic inhibition of LTßR signalling in young and aged mice disrupted smoking-related inducible bronchus-associated lymphoid tissue, induced regeneration of lung tissue, and reverted airway fibrosis and systemic muscle wasting. Mechanistically, blockade of LTßR signalling dampened epithelial non-canonical activation of NF-κB, reduced TGFß signalling in airways, and induced regeneration by preventing epithelial cell death and activating WNT/ß-catenin signalling in alveolar epithelial progenitor cells. These findings suggest that inhibition of LTßR signalling represents a viable therapeutic option that combines prevention of tertiary lymphoid structures1 and inhibition of apoptosis with tissue-regenerative strategies.

Environmental salinity modulates olfactory sensitivity in the euryhaline European seabass, Dicentrarchus labrax, acclimated to seawater and brackish water.

The olfactory epithelium of fish is - of necessity - in intimate contact with the surrounding water. In euryhaline fish, movement from seawater to freshwater (and vice versa) exposes the epithelium to massive changes in salinity and ionic concentrations. How does the olfactory system function in the face of such changes? The current study compared olfactory sensitivity in seawater- (35‰) and brackish water-adapted seabass (5‰) using extracellular multi-unit recording from the olfactory nerve. Seawater-adapted bass had higher olfactory sensitivity to amino acid odorants when delivered in seawater than in freshwater. Conversely, brackish water-adapted bass had largely similar sensitivities to the same odorants when delivered in seawater or freshwater, although sensitivity was still slightly higher in seawater. The olfactory system of seawater-adapted bass was sensitive to decreases in external [Ca2+], whereas brackish water-adapted bass responded to increases in [Ca2+]; both seawater- and brackish water-adapted bass responded to increases in external [Na+] but the sensitivity was markedly higher in brackish water-adapted bass. In seawater-adapted bass, olfactory sensitivity to l-alanine depended on external Ca2+ ions, but not Na+; brackish water-adapted bass did respond to l-alanine in the absence of Ca2+, albeit with lower sensitivity, whereas sensitivity was unaffected by removal of Na+ ions. A possible adaptation of the olfactory epithelium was the higher number of mucous cells in brackish water-adapted bass. The olfactory system of seabass is able to adapt to low salinities, but this is not immediate; further studies are needed to identify the processes involved.

Anatomy of the olfactory system and potential role for chemical communication in the sound-producing Lusitanian toadfish, Halobatrachus didactylus.

The current study investigated the structure and function of the olfactory system of the Lusitanian toadfish, Halobatrachus didactylus, using histology and electrophysiology (electro-olfactogram [EOG]), respectively. The olfactory system consists of a digitated anterior peduncle, of unknown function, containing the inhalant nostril. This then leads to a U-shaped olfactory chamber with the olfactory epithelium-identified by Gαolf-immunoreactivity-on the ventral surface. A large lacrimal sac is connected to this tube and is likely involved in generating water movement through the olfactory chamber (this species is largely sedentary). The exhalent nostril lies by the eye and is preceded by a bicuspid valve to ensure one-way flow of water. As do other teleosts, H. didactylus had olfactory sensitivity to amino acids and bile acids. Large-amplitude EOG responses were evoked by fluid from the anterior and posterior testicular accessory glands, and bile and intestinal fluids. Anterior gland and intestinal fluids from reproductive males were significantly more potent than those from non-reproductive males. Male urine and skin mucus proved to be the least potent body fluids tested. These results suggest that chemical communication-as well as acoustic communication-may be important in the reproduction of this species and that this may be mediated by the accessory glands and intestinal fluid.

Impact of disease duration on proteomic bioprofile and prognosis in rheumatoid arthritis patients.

OBJECTIVES: Cardiovascular disease worsens the prognosis of rheumatoid arthritis (RA) and vice-versa. Inflammation may be a common pathway for both conditions. It is expected that a longer RA duration leads to a greater inflammatory cumulative exposure burden; however, studies on the association between RA disease duration and outcomes are scarce. Our aim is to compare the characteristics, biomarker expression and outcomes according to the duration of RA. METHODS: Prospective cohort study including 399 RA patients, with detailed clinical, echocardiographic, and proteomic phenotyping that were compared across tertiles of RA disease duration. Cox proportional models were used to study the association of disease duration with cardiovascular outcomes. RESULTS: RA duration tertiles were: tertile 1 with median of 3.2; tertile 2 with median of 8.8; and tertile 3 with median of 21.8 years. Compared to tertile 1, patients in tertile 3 were older, had more erosive disease, more frequent echocardiographic alterations, lower haemoglobin and walked a shorter distance on the 6MWT. Natriuretic peptides, cathepsin L1, galectin 9, matrix metalloproteinase-12, adrenomedullin and tumour necrosis factor receptor 11A were higher in patients with longer disease duration. Compared to patients in tertile 1, those in tertile 3 had higher risk of a subsequent cardiovascular hospitalisation or cardiovascular death (HR 2.71, 95%CI 1.06-6.92, p=0.04). CONCLUSIONS: RA patients with longer disease duration had more organ damage and worse outcomes than those with shorter disease duration. Biomarker expression suggested that patients with longer RA duration had activation of pathways related to inflammation, extracellular matrix organisation, fibrosis and congestion.

Thermal imprinting during embryogenesis modifies skin repair in juvenile European sea bass (Dicentrarchus labrax).

Fish skin is a multifunctional tissue that develops during embryogenesis, a developmental stage highly susceptible to epigenetic marks. In this study, the impact of egg incubation temperature on the regeneration of a cutaneous wound caused by scale removal in juvenile European sea bass was evaluated. Sea bass eggs were incubated at 11, 13.5 and 16 °C until hatching and then were reared at a common temperature until 9 months when the skin was damaged and sampled at 0, 1 and 3 days after scale removal and compared to the intact skin from the other flank. Skin damage elicited an immediate significant (p < 0.001) up-regulation of pcna in fish from eggs incubated at higher temperatures. In fish from eggs incubated at 11 °C there was a significant (p < 0.001) up-regulation of krt2 compared to fish from higher thermal backgrounds 1 day after skin damage. Damaged epidermis was regenerated after 3 days in all fish irrespective of the thermal background, but in fish from eggs incubated at 11 °C the epidermis was significantly (p < 0.01) thinner compared to other groups, had less goblet cells and less melanomacrophages. The thickness of the dermis increased during regeneration of wounded skin irrespective of the thermal background and by 3 days was significantly (p < 0.01) thicker than the dermis from the intact flank. The expression of genes for ECM remodelling (mmp9, colXα, col1α1, sparc, and angptl2b) and innate immunity (lyg1, lalba, sod1, csf-1r and pparγ) changed during regeneration but were not affected by egg thermal regime. Overall, the results indicate that thermal imprinting of eggs modifies the damage-repair response in juvenile sea bass skin.

When benign leiomyomas metastasize to the lungs - a case report.

Benign metastasizing leiomyoma is a rare disease that primarily affects premenopausal women who have a history of uterine leiomyoma, which is characterized by pulmonary metastases. The pathogenesis of this condition is unknown. Patients are usually asymptomatic or have vague symptoms. Pathological examination in conjunction with immunohistochemistry is required for diagno-sis. Treatment is determined by the patient's age, hormonal status, symptoms, and the extent of the lesions (number, size, and location of nodules), with surgical resection being the most effective. We present the case of a 72-year-old woman who had a total hysterec-tomy 30 years earlier due to myomas and developed a persistent dry cough. Her computed tomography of the chest revealed several small nodules in the pulmonary parenchyma. Because of the nod-ules' small size and difficult accessibility, a surgical biopsy was per-formed for histopathological examination and immunohistochemi-cal staining, which revealed metastasizing leiomyoma. Letrozole treatment was started and was well tolerated.

Endoscopic ampullectomy of a giant pedunculated ampullary adenoma.

The authors present a case of a 72-year-old woman with a personal history of arterial hypertension, dyslipidemia and gallstones. For suspected choledocholithiasis, a MR-Cholangiopancreatography was performed, which revealed the presence of an endoluminal polypoid structure in the third duodenal portion.

Gastric mixed neuroendocrine/non neuroendocrine neoplasia: a rare entity.

Gastric neoplasms have different histological subtypes. Mixed neuroendocrine/non neuroendocrine neoplasms (MiNEN), as defined by the World Health Organization (WHO) 2019 Classification of Tumours of the Digestive System, are rare composite tumours bearing morphological characteristics of more than one histological type. Historically, the diagnosis requires that each component accounts for at least 30% of the total tumour burden. We present the case of a 70-year-old male, who was referred to our endoscopy unit for resection of a depressed lesion (Paris classification type 0 IIc) of 40mm in largest diameter, located on the lesser curvature of the gastric body. The lesion was submitted to en bloc resection by endoscopic submucosal dissection. Histological examination revealed a poorly differentiated intramucosal adenocarcinoma and a grade G3 well differentiated neuroendocrine tumour (NET), on a background of chronic atrophic gastritis. Unfortunately, examination of the NET component revealed intercepted margins. The patient is currently undergoing tumour staging and will be discussed by a multidisciplinary team to determine subsequent management.

Variants in ADD1 cause intellectual disability, corpus callosum dysgenesis, and ventriculomegaly in humans.

PURPOSE: Adducins interconnect spectrin and actin filaments to form polygonal scaffolds beneath the cell membranes and form ring-like structures in neuronal axons. Adducins regulate mouse neural development, but their function in the human brain is unknown. METHODS: We used exome sequencing to uncover ADD1 variants associated with intellectual disability (ID) and brain malformations. We studied ADD1 splice isoforms in mouse and human neocortex development with RNA sequencing, super resolution imaging, and immunoblotting. We investigated 4 variant ADD1 proteins and heterozygous ADD1 cells for protein expression and ADD1-ADD2 dimerization. We studied Add1 functions in vivo using Add1 knockout mice. RESULTS: We uncovered loss-of-function ADD1 variants in 4 unrelated individuals affected by ID and/or structural brain defects. Three additional de novo copy number variations covering the ADD1 locus were associated with ID and brain malformations. ADD1 is highly expressed in the neocortex and the corpus callosum, whereas ADD1 splice isoforms are dynamically expressed between cortical progenitors and postmitotic neurons. Human variants impair ADD1 protein expression and/or dimerization with ADD2. Add1 knockout mice recapitulate corpus callosum dysgenesis and ventriculomegaly phenotypes. CONCLUSION: Our human and mouse genetics results indicate that pathogenic ADD1 variants cause corpus callosum dysgenesis, ventriculomegaly, and/or ID.

GABA receptors in the olfactory epithelium of the gilthead seabream (Sparus aurata).

Exposure to high PCO2/low pH seawater induces behavioural alterations in fish; a possible explanation for this is a reversal of Cl-/HCO3- currents through GABAA receptors (the GABAA receptor theory). However, the main evidence for this is that gabazine, a GABAA receptor antagonist, reverses these effects when applied to the water, assuming that exposure to systems other than the CNS would be without effect. Here, we show the expression of both metabotropic and ionotropic GABA receptors, and the presence of GABAA receptor protein, in the olfactory epithelium of gilthead seabream. Furthermore, exposure of the olfactory epithelium to muscimol (a specific GABAA receptor agonist) increases or decreases the apparent olfactory sensitivity to some odorants. Thus, although the exact function of GABAA receptors in the olfactory epithelium is not yet clear, this may complicate the interpretation of studies wherein water-borne gabazine is used to reverse the effects of high CO2 levels on olfactory-driven behaviour in fish.

Predictors of post-COVID syndrome. Getting ready for the future.

Dear Editor, We have read with interest the study by Gulian et al.  where the authors describe the impact of prolonged symptoms or developing complications following an initial recovery from COVID-19, also called post-COVID syndrome. In this aspect there are a small amount of studies based on clinical extrapolation....

Independent effects of seawater pH and high PCO2 on olfactory sensitivity in fish: possible role of carbonic anhydrase.

Ocean acidification may alter olfactory-driven behaviour in fish by direct effects on the peripheral olfactory system; olfactory sensitivity is reduced in CO2-acidified seawater. The current study tested whether this is due to elevated PCO2  or the consequent reduction in seawater pH and, if the former, the possible involvement of carbonic anhydrase, the enzyme responsible for the hydration of CO2 and production of carbonic acid. Olfactory sensitivity to amino acids was assessed by extracellular multi-unit recording from the olfactory nerve of the gilthead seabream (Sparus aurata L.) in normal seawater (pH ∼8.2), and after acute exposure to acidified seawater (pH ∼7.7) but normal PCO2  (∼340 µatm) or to high PCO2  seawater (∼1400 µatm) at normal pH (∼8.2). Reduced pH in the absence of elevated PCO2  caused a reduction in olfactory sensitivity to l-serine, l-leucine, l-arginine and l-glutamine, but not l-glutamic acid. Increased PCO2  in the absence of changes in pH caused reduced olfactory sensitivity to l-serine, l-leucine and l-arginine, including increases in their threshold of detection, but had no effect on sensitivity to l-glutamine and l-glutamic acid. Inclusion of 1 mmol l-1 acetazolamide (a membrane-permeant inhibitor of carbonic anhydrase) in the seawater reversed the inhibition of olfactory sensitivity to l-serine caused by high PCO2 Ocean acidification may reduce olfactory sensitivity by reductions in seawater pH and intracellular pH (of olfactory receptor neurones); the former by reducing odorant-receptor affinity, and the latter by reducing the efficiency of olfactory transduction. The physiological role of carbonic anhydrase in the olfactory receptor neurones remains to be explored.

A Histopathological Study Reveals New Insights Into Responses of Chestnut (Castanea spp.) to Root Infection by Phytophthora cinnamomi.

The European chestnut (Castanea sativa) is threatened by the hemibiotrophic oomycete Phytophthora cinnamomi, the causal agent of ink disease. Chestnut species have different susceptibility levels to P. cinnamomi, with the Asian species (C. crenata; C. mollissima) exhibiting the highest level of resistance. A histological approach was used to study the responses exhibited by susceptible and resistant chestnut genotypes by characterizing the early stages of P. cinnamomi infection and the cellular responses it induces in roots. C. sativa (susceptible) and C. crenata (resistant) plantlets were inoculated with a P. cinnamomi virulent isolate with a zoospore suspension or by direct contact with mycelia agar pieces. Root samples were collected at 0.5, 3.5, 24, 48, and 72 h after inoculation (hai) for microscopic observations. Penetration was observed in both species at 0.5 and 3.5 hai with mycelium and zoospore inoculations, respectively. In both inoculation methods, following penetration into the rhizodermis, P. cinnamomi hyphae grew inter- and intracellularly through the cortex and into the vascular cylinder. C. crenata cells displayed a delay in the pattern of infection by having fewer cell layers colonized compared with C. sativa. At 72 hai, the collapse of the first layers of C. sativa cortical cells was observed, indicating the beginning of necrotrophy. C. crenata was able to respond more efficiently to P. cinnamomi than C. sativa by restricting the pathogen's growth area through the early activation of resistance responses such as callose deposition around some intracellular hyphae, hypersensitive response-like cell death, cell wall thickening, and accumulation of phenolic-like compounds.

Cartilage Acidic Protein a Novel Therapeutic Factor to Improve Skin Damage Repair?

Fish skin has been gaining attention due to its efficacy as a human-wound-treatment product and to identify factors promoting its enhanced action. Skin fibroblasts have a central role in maintaining skin integrity and secrete extra cellular matrix (ECM) proteins, growth factors and cytokines to rapidly repair lesions and prevent further damage or infection. The effects on scratch repair of the ubiquitous but poorly characterized ECM protein, cartilage acidic protein 1 (CRTAC1), from piscine and human sources were compared using a zebrafish SJD.1 primary fibroblast cell line. A classic in vitro cell scratch assay, immunofluorescence, biosensor and gene expression analysis were used. Our results demonstrated that the duplicate sea bass Crtac1a and Crtac1b proteins and human CRTAC-1A all promoted SJD.1 primary fibroblast migration in a classic scratch assay and in an electric cell impedance sensing assay. The immunofluorescence analysis revealed that CRTAC1 enhanced cell migration was most likely caused by actin-driven cytoskeletal changes and the cellular transcriptional response was most affected in the early stage (6 h) of scratch repair. In summary, our results suggest that CRTAC1 may be an important factor in fish skin promoting damage repair.

Cardiothoracic surgery during the Covid-19 pandemic: Perioperative care, safety, and surgical results.

BACKGROUND: Covid-19 was declared a pandemic by the World Health Organization (WHO) on 11 March 2020. Responses to this crisis integrated resource allocation for the increased amount of infected patients while maintaining an adequate response to other severe and life-threatening diseases. Though cardiothoracic patients are at high risk for Covid-19 severe illness, postponing surgeries would translate to increased mortality and morbidity. We reviewed our practice during the initial time of the pandemic, with emphasis on safety protocols. METHODS: From 11 March to 15 May 2020, 148 patients underwent surgery at the Department of Cardiothoracic Surgery of Centro Hospitalar Universitário de São João. The clinical characteristics of the patients were retrospectively registered, along with novel containment and infection prevention measures targeting the new coronavirus. RESULTS: The majority of adult cardiac patients were operated on an emergency basis. Hospital mortality was 1.9% (n = 2 patients). Most adult thoracic patients were admitted from home, with a diagnosis of neoplastic disease in 60% of patients. Hospital mortality was 3.3% (1 patient). Fifteen children underwent cardiothoracic surgery. There was no mortality. The infection prevention procedures applied, totally excluded the transmission of Covid-19 in the department. CONCLUSION: While guaranteeing a prompt response to emergent, urgent, and high priority cases, novel safety measures in individual protection, patients circuits, and preoperative diagnoses of symptomatic and asymptomatic infection were adopted. The surgical results corroborate that it was safe to undergo cardiothoracic surgery during the initial time of the Covid-19 pandemic. The new policies will be maintained while the virus stays in the community.

Embolization Of Implanon Devices Lung Sparing Videoassisted Thoracic Surgery.

A 31-year-old-woman with an etonogestrel implant on her left upper arm presented with unfavorable change in her menstrual bleeding pattern and requested for its removal. The non-palpable device was perceptible in the left hemithorax by radiography. Thoracic computed-tomography showed migration to a sublobar branch of the left lower pulmonary artery. Despite the absence of thoracic symptoms and the lack of management guidelines, the device was removed by a lung sparing approach with videoassisted thoracic surgery, due to the unknown long-term effect of the embolized implant.

Long-term follow-up after EMR of large colorectal lesions: metachronous lesions, predictors and surveillance timing.

Background: Endoscopic mucosal resection (EMR) is the first-line approach to large colorectal sessile lesions. These patients have been associated with high rates of metachronous lesions (ML), but long-term follow-up (LtFU) data are lacking. We aimed at evaluating the efficacy of an LtFU protocol and analyse the development and risk factors for ML.Methods: A prospectively collected database was analysed. Seventy-six patients submitted to EMR of large colorectal sessile lesions between 2007 and 2013 complied with a specific endoscopic surveillance, consisting of two protocols - initial follow-up (iFU) and LtFU. iFU intended to inspect the mucosectomy scars twice (at 3-6 and 12 months) and remove synchronous lesions (SL). Protocol examinations of LtFU were carried out at the first- and fourth-year post-iFU, aiming to remove ML. Statistical analysis included variables related to patient, index lesion, SL and ML characteristics.Results: Rates of ML were 39.5% and 20.4% at the first- and fourth-year of LtFU, and respectively 11.8% and 3.7% of them were advanced ML. All ML were endoscopically resectable. At univariate analysis, male gender (OR: 2.91; p=.029), the presence of SL (OR 3.86, p=.010) and advanced SL (OR 4.25, p=.006) were risk factors for ML. At multivariate analysis, male gender (p=.031) and advanced SL (p=.006) were significant predictors of ML development.Conclusions: We confirmed the increased risk of ML in patients with large colorectal lesions. A significant number of advanced ML was removed at the first LtFU colonoscopy, probably it should be carried out earlier than currently recommended.

Mirizzi syndrome: when the gallbladder meets bile ducts.

A 64-year-old female presented to the Emergency Department with jaundice, choluria, fever and abdominal pain over the last few days. The abdomen was tender with epigastric pain on palpation.

Lung Hernia Related with a Rope Bullfight: Case Report.

Lung herniation is an uncommon entity which was fully classified in 1845 after the study of several case reports. Acquired lung hernia, especially traumatic, is the most common etiology. In the absence of clear guidelines, management of lung hernia is made in a case-by-case basis. We present an asymptomatic middle lobe hernia perceptible on physical examination, but diagnosed initially by imaging studies. Patient medical history included a blunt bull trauma fourteen years before.