Middle East respiratory syndrome coronavirus (MERS-CoV) infections in two returning travellers in the Netherlands, May 2014

Two patients, returning to the Netherlands from pilgrimage in Medina and Mecca, Kingdom of Saudi Arabia, were diagnosed with Middle East respiratory syndrome coronavirus (MERS-CoV) infection in May 2014. The source and mode of transmission have not yet been determined. Hospital-acquired infection and community-acquired infection are both possible.

Rôle des ports dans le cycle de vie des poissons. Les ports : nurseries de sars ?

Le présent document constitue un rapport final de l’étude dont l’objectif peut s’énoncer comme suit : « les zones portuaires peuvent elles être des nurseries de sars ? ». Tenter de répondre à cette question, simple en apparence, nécessite de mener des études aux niveaux populationnel et individuel et ce afin de comprendre les processus sous-jacents. Quatre volets ont été menés. Dans le premier volet, à partir des données issues du projet Nappex, nous avons étudié les changements spatio-temporels d’abondance de quatre espèces de sars dans cinq marinas réparties le long de la façade Méditerranéenne. Dans un second volet, nous avons cherché à comparer les cinétiques de mortalité post-installation de deux espèces de sars en milieu portuaire et naturel. Dans un troisième volet, la croissance et la condition de deux mêmes espèces de sars ont été comparées entre des zones portuaires et des zones naturelles. Enfin, dans un quatrième volet, nous avons cherché à mettre en place des outils pour appréhender la migration des juvéniles des zones portuaires vers les zones naturelles pour rejoindre les populations adultes.

New cholinesterase inhibitors for Alzheimer's disease: Structure Activity Studies (SARs) and molecular docking of isoquinolone and azepanone derivatives

A library of isoquinolinone and azepanone derivatives were screened for both acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) activity. The strategy adopted included (a) in vitro biological assays, against eel AChE (EeAChE) and equine serum BuChE (EqBuChE) in order to determine the compounds IC50 and their dose-response activity, consolidated by (b) molecular docking studies to evaluate the docking poses and interatomic interactions in the case of the hit compounds, validated by STD-NMR studies. Compound (1f) was identified as one of these hits with an IC50 of 89.5 mu M for EeAChE and 153.8 mu M for EqBuChE, (2a) was identified as a second hit with an IC50 of 108.4 mu M (EeAChE) and 277.8 mu M (EqBuChE). In order to gain insights into the binding mode and principle active site interactions of these molecules, (R)-(1f) along with 3 other analogues (also as the R-enantiomer) were docked into both RhAChE and hBuChE models. Galantamine was used as the benchmark. The docking study was validated by performing an STD-NMR study of (1f) with EeAChE using galantamine as the benchmark.

Reti neurali "physics-informed" per modelli epidemiologici

La malattia COVID-19 associata alla sindrome respiratoria acuta grave da coronavirus 2 (SARS-CoV-2) ha rappresentato una grave minaccia per la salute pubblica e l’economia globale sin dalla sua scoperta in Cina, nel dicembre del 2019. Gli studiosi hanno effettuato numerosi studi ed in particolar modo l’applicazione di modelli epidemiologici costruiti a partire dai dati raccolti, ha permesso la previsione di diversi scenari sullo sviluppo della malattia, nel breve-medio termine. Gli obiettivi di questa tesi ruotano attorno a tre aspetti: i dati disponibili sulla malattia COVID-19, i modelli matematici compartimentali, con particolare riguardo al modello SEIJDHR che include le vaccinazioni, e l’utilizzo di reti neurali ”physics-informed” (PINNs), un nuovo approccio basato sul deep learning che mette insieme i primi due aspetti. I tre aspetti sono stati dapprima approfonditi singolarmente nei primi tre capitoli di questo lavoro e si sono poi applicate le PINNs al modello SEIJDHR. Infine, nel quarto capitolo vengono riportati frammenti rilevanti dei codici Python utilizzati e i risultati numerici ottenuti. In particolare vengono mostrati i grafici sulle previsioni nel breve-medio termine, ottenuti dando in input dati sul numero di positivi, ospedalizzati e deceduti giornalieri prima riguardanti la città di New York e poi l’Italia. Inoltre, nell’indagine della parte predittiva riguardante i dati italiani, si è individuato un punto critico legato alla funzione che modella la percentuale di ricoveri; sono stati quindi eseguiti numerosi esperimenti per il controllo di tali previsioni.

Tecniche di group testing per il rilevamento di SARS-CoV-2

Oggigiorno l'individuazione diagnostica precoce della SARS-CoV-2 attraverso tamponi molecolari è fondamentale per interrompere la trasmissione del virus. Tuttavia, il monitoraggio della diffusione virale attraverso il test standard di RT-qPCR individuale comporta un elevato costo per ciascun tampone nasofaringeo analizzato e i reagenti chimici per l’estrazione dell’RNA virale sono sempre meno disponibili. Per ovviare a tali ostacoli, è stata ripresa la tecnica di group testing, sviluppata per la prima volta da Dorfman nel 1943 per individuare i soggetti affetti da sifilide prima del loro arruolamento. Questa strategia minimizza il numero di test condotti su un insieme di campioni: se un gruppo di n campioni risulta negativo, allora la condizione di ciascuno di essi è stata determinata mediante un solo test invece che con n test individuali. Negli ultimi due anni sono state sviluppate strategie in grado di migliorare le prestazioni del test di gruppo: per scenari a bassa prevalenza l’algoritmo dell’ipercubo rileva un singolo campione positivo in pool con dimensioni fino a 100 campioni attraverso due o più turni di test; invece, il P-BEST utilizza un solo turno di analisi, ma le dimensioni massime dei pool sono più ridotte. Per scenari ad alta prevalenza (10%) il team italiano dell’Università di Bologna ha progettato un metodo che identifica e rileva i membri infetti con un solo turno di test. Tuttavia, affinché il group testing sia efficace come l’analisi individuale dei tamponi molecolari, è necessario minimizzare l’effetto di diluizione, correlato alla dimensione del pool e causa di insorgenza di falsi negativi, nonché di un calo nella sensibilità nei test. I dati ottenuti da questi studi hanno dimostrato che questa strategia offre grandi potenzialità. Essa è essenziale per le indagini di routine della popolazione e concede vantaggi amplificati soprattutto se vengono testati soggetti quotidianamente in contatto tra loro, come famiglie o colleghi di lavoro.

Achados clínico patológicos durante um surto de disenteria de inverno em bovinos no Rio Grande do Sul

Descreve-se um surto de disenteria de inverno que afetou 10 vacas leiteiras de uma propriedade localizada em Viamão, Rio Grande do Sul. O quadro clínico caracterizou-se por uma diarréia inicialmente líquida esverdeada com estrias de sangue e muco, evoluindo, em alguns animais, para uma diarréia de coloração marrom escura à sanguinolenta, que persistiu, em média, cinco dias. Drástica diminuição na produção de leite e no consumo de alimentos, além de graus variados de depressão também foram observados. Apenas um dos 10 animais afetados morreu. Durante a necropsia, observaram-se mucosas pálidas, conteúdo sanguinolento com presença de grande quantidade de coágulos, principalmente no cólon espiral e petéquias na mucosa do cólon. Os principais achados histológicos foram encontradas no cólon espiral, onde havia criptas dilatadas, sem epitélio de revestimento ou revestidas por epitélio pavimentoso e/ou cuboidal, por vezes com núcleos grandes e nucléolos proeminentes. Algumas criptas eram preenchidas por debris necróticos e polimorfonucleares. Na imuno-histoquímica com anticorpo monoclonal para coronavírus bovino (8F2) em cortes do cólon espiral, havia marcações positivas no citoplasma de enterócitos das criptas, nos debris necróticos destas e em macrófagos na lâmina própria.

Genetically Diverse Coronaviruses in Captive Bird Populations in a Brazilian Zoological Park

This study aimed to investigate the occurrence of coronaviruses (CoVs) in captive birds placed inside a zoological park in Brazil. The role of captive birds in the epidemiology of CoVs in the tropics is poorly understood. A total of 25 (n = 25) different species were tested for viral RNA using individual fecal samples collected from healthy birds. Reverse transcription-polymerase chain reaction targeting the 30 untranslated region was used to detect CoV RNA, and positive samples were submitted for sequence analysis. The phylogenetic search revealed nine mutations in the black shouldered peafowl (Pavus cristatus) CoV sequence, which clustered separately from samples previously described in England. This is the first report on the detection of the CoV genome in captive birds in Brazil.

Cryptic species, life cycles, and the phylogeny of Clytia (Cnidaria: Hydrozoa: Campanulariidae)

Medusae and polyps of Clytia are abundantly found in coastal marine environments and one species in the genus-Clytia hemisphaerica (Linnaeus, 1767)-has become an important experimental model. Yet, only 10 species in the genus have had their life cycle investigated. Most species of Clytia are also poorly described, and detailed life cycle and morphological studies are needed for accurate species-level identifications. Here, we investigated the life cycle of Clytia elsaeoswaldae Stechow, 1914, a species described for the tropical western Atlantic and subsequently considered conspecific to the nearly-cosmopolitan species Clytia gracilis (Sars, 1850) and Clytia hemisphaerica, originally described for the temperate North Atlantic. Based on observations of mature medusae and multiple colonies from southeastern Brazil and the U. S. Virgin Islands (type locality), our results show that C. elsaeoswaldae is morphologically distinct from C. gracilis and C. hemisphaerica. The morphological results are corroborated by a multigene phylogenetic analysis of the genus Clytia, which shows that C. gracilis-like species form a polyphyletic group of several species. These results suggest that the nearly-cosmopolitan distribution attributed to some species of Clytia may be due to the non-recognition of morphologically similar species with more restricted ranges.

Wild canids, domestic dogs and their pathogens in Southeast Brazil: disease threats for canid conservation

Wild canids are under many pressures, including habitat loss, fragmentation and disease. The current lack of information on the status of wildlife health may hamper conservation efforts in Brazil. In this paper, we examined the prevalence of canine pathogens in 21 free-ranging wild canids, comprising 12 Cerdocyon thous (crab-eating fox), 7 Chrysocyon brachyurus (maned wolf), 2 Lycalopex vetulus (hoary fox), and 70 non-vaccinated domestic dogs from the Serra do Cip National Park area, Southeast Brazil. For wild canids, seroprevalence of antibodies to canine parvovirus, canine adenovirus, canine coronavirus and Toxoplasma gondii was 100 (21/21), 33 (7/21), 5 (1/19) and 68 (13/19) percent, respectively. Antibodies against canine distemper virus, Neospora caninum or Babesia spp. were not found. We tested domestic dogs for antibodies to canine parvovirus, canine distemper virus and Babesia spp., and seroprevalences were 59 (41/70), 66 (46/70), and 42 (40/70) percent, respectively, with significantly higher prevalence in domestic dogs for CDV (P < 0.001) and Babesia spp. (P = 0.002), and in wild canids for CPV (P < 0.001). We report for the first time evidence of exposure to canine coronavirus in wild hoary foxes, and Platynossomun sp. infection in wild maned wolves. Maned wolves are more exposed to helminths than crab-eating foxes, with a higher prevalence of Trichuridae and Ancylostomidae in the area. The most common ectoparasites were Amblyomma cajennense, A. tigrinum, and Pulex irritans. Such data is useful information on infectious diseases of Brazilian wild canids, revealing pathogens as a threat to wild canids in the area. Control measures are discussed.

SURVEY OF FELINE LEUKEMIA VIRUS AND FELINE CORONAVIRUSES IN CAPTIVE NEOTROPICAL WILD FELIDS FROM SOUTHERN BRAZIL

A total of 57 captive neotropical felids (one Leopardus geoffroyi, 14 Leopardus pardalis, 17 Leopardus wiedii, 22 Leopardus tigrinus, and three Puma yagouaroundi) from the Itaipu Binacional Wildlife Research Center (Refugio Bela Vista, Southern Brazil) were anesthetized for blood collection. Feces samples were available for 44 animals, including one L. geoffroyi, eight L. pardalis, 14 L. wiedii, 20 L. tigrinus, and one P. yagouaroundi. Total DNA and RNA were extracted from blood and feces, respectively, using commercial kits. Blood DNA samples were evaluated by polymerase chain reaction (PCR) for feline leukemia virus (FeLV) proviral DNA, whereas reverse transcriptase-PCR was run on fecal samples for detection of coronavirus RNA. None of the samples were positive for coronaviruses. A male L. pardalis and a female L. tigrinus were positive for FeLV proviral DNA, and identities of PCR products were confirmed by sequencing. This is the first evidence of FeLV proviral DNA in these species in Southern Brazil.

Cloning and functional analysis of the Sry-related HMG box gene, Sox18

The Sox gene family (Sry like HMG box gene) is characterised by a conserved DNA sequence encoding a domain of approximately 80 amino acids which is responsible for sequence specific DNA binding. We initially published the identification and partial cDNA sequence of murine Sox18, a new member of this gene family, isolated from a cardiac cDNA library. This sequence allowed us to classify Sox18 into the F sub-group of Sox proteins, along with Sox7 and Sox17. Recently, we demonstrated that mutations in the Sox18 activation domain underlie cardiovascular and hair follicle defects in the mouse mutation, ragged (Ra) (Pennisi et al., 2000. Mutations in Sox18 underlie cardiovascular and hair follicle defecs in ragged mice. Nat. Genet. 24, 434-437). Ra homozygotes lack vibrissae and coat hairs, have generalised oedema and an accumulation of chyle in the peritoneum. Here we have investigated the genomic sequences encoding Sox18. Screening of a mouse genomic phage library identified four overlapping clones, we sequenced a 3.25 kb XbaI fragment that defined the entire coding region and approximately 1.5 kb of 5' flanking sequences. This identified (i) an additional 91 amino acids upstream of the previously designated methionine start codon in the original cDNA, and (ii);ln intron encoded within the HMG box/DNA binding domain in exactly the same position as that found in the Sox5, -13 and -17 genes. The Sox18 gene encodes a protein of 468 aa. We present evidence that suggests HAF-2, the human HMG-box activating factor-2 protein, is the orthologue of murine Sox18. HAF-2 has been implicated in the regulation of the Human IgH enhancer in a B cell context. Random mutagenesis coupled with GAL4 hybrid analysis in the activation domain between amino acids 252 and 346, of Sox18, implicated the phosphorylation motif, SARS, and the region between amino acid residues 313 and 346 as critical components of Sox18 mediated transactivation. Finally, we examined the expression of Sox18 in multiple adult mouse tissues using RT-PCR. Low-moderate expression was observed in spleen, stomach, kidney, intestine, skeletal muscle and heart. Very abundant expression was detected in lung tissue. (C) 2001 Elsevier Science B.V. All rights reserved.

Point source inoculation of Mesocyclops (Copepoda : Cyclopidae) gives widespread control of Ochlerotatus and Aedes (Diptera : Culicidae) immatures in service manholes and pits in North Queensland, Australia

This study details the novel application of predacious copepods, genus Mesocyclops, for control of Ochlerotatus tremulus (Theobald) group and Aedes aegypti (L.) mosquito larvae in subterranean habitats in north Queensland, Australia. During June 1997, 50 Mesocyclops sp. I were inoculated into one service manhole in South Townsville. Wet season rainfall and flooding in both 1998 and 2000 was responsible for the dispersal of copepods via the underground pipe system to 29 of 35 manholes over an area of 1.33 km(2). Significant reductions in Aedes and Ochlerotatus larvae ensued. In these habitats, Mesocyclops and Metacyclops were able to survive dry periods, when substrate moisture content ranged from 13.8 to 79.9%. At the semiarid inland towns of Hughenden and Richmond, cracking clay soil prevents drainage of water from shallow service pits where Oc. tremulus immatures numbered from 292-18,460 per pit. Introduction of Mesocyclops copepods into these sites during May 1999 resulted in 100% control of Oc. tremulus for 18 mo. One uninoculated pit subsequently became positive for Mesocyclops with resultant control of mosquito larvae.

Telehealth responses to bio-terrorism and emerging infections

Emerging infectious diseases, such as severe acute respiratory syndrome (SARS), are of huge economic importance. They are difficult to predict. The World Health Organization has a Global Outbreak Alert and Response Network, which was involved at an early stage in the SARS outbreak in 2003. Three major lessons were learned as a result of the SARS epidemic in 2003, involving communication, evidence-based action and global partnerships. It is proposed that a series of broadband global response networks should be developed. At a technical level the networks are essentially in place, such as the Internet2 global network. Suitable peripheral devices also exist. What has not yet been created is the appropriate software to allow the use of these networks, although a number of commercial products are in the process of development.

Antioxidant properties of hydroxycinnamic acids: a review of structure- activity relationships

Hydroxycinnamic acids (HCAs) are important phytochemicals possessing significant biological properties. Several investigators have studied in vitro antioxidant activity of HCAs in detail. In this review, we have gathered the studies focused on the structure-activity relationships (SARs) of these compounds that have used medicinal chemistry to generate more potent antioxidant molecules. Most of the reports indicated that the presence of an unsaturated bond on the side chain of HCAs is vital to their activity. The structural features that were reported to be of importance to the antioxidant activity were categorized as follows: modifications of the aromatic ring, which include alterations in the number and position of hydroxy groups and insertion of electron donating or withdrawing moieties as well as modifications of the carboxylic function that include esterification and amidation process. Furthermore, reports that have addressed the influence of physicochemical properties including redox potential, lipid solubility and dissociation constant on the antioxidant activity were also summarized. Finally, the pro-oxidant effect of HCAs in some test systems was addressed. Most of the investigations concluded that the presence of ortho-dihydroxy phenyl group (catechol moiety) is of significant importance to the antioxidant activity, while, the presence of three hydroxy groups does not necessarily improve the activity. Optimization of the structure of molecular leads is an important task of modern medicinal chemistry and its accomplishment relies on the careful assessment of SARs. SAR studies on HCAs can identify the most successful antioxidants that could be useful for management of oxidative stress-related diseases.

Murine virus contaminant of Trypanosoma cruzi experimental infection

The possibility that some virus contaminants could be altering host response to Trypanosoma cruzi experimental infection was investigated. Data obtained showed that CBA/J mice infected with stocks of parasite maintained in mice (Y1UEC) presented higher level of parasitemia and shorter survival times than those infected with a stock (Y1TC) which was also maintained in mice but had been previously passaged in cell culture. Mouse antibody production tests, performed with the filtered plasma of mice infected with Y1UEC, indicated the presence of mouse hepatitis virus (MHV) while no virus was detected when testing the plasma of Y1TC infected mice. Filtered plasma of Y1EUC infected mice was shown to contain a factor able to enhance the level of parasitemia and to reduce the mean survival time of mice challenged with 10(5) Y1TC. This factor, that could be serially passaged to naïve mice was shown to be a coronavirus by neutralization tests.