Mutations in the Schmallenberg virus Gc glycoprotein facilitate cellular protein synthesis shutoff and restore pathogenicity of NSs deletion mutants in mice

Serial passage of viruses in cell culture has been traditionally used to attenuate virulence and identify determinants of viral pathogenesis. In a previous study, we found that a strain of Schmallenberg virus (SBV) serially passaged in tissue culture (termed SBVp32) unexpectedly displayed increased pathogenicity in suckling mice compared to wild type SBV. In this study, we mapped the determinants of SBVp32 virulence to the viral genome M segment. SBVp32 virulence is associated with the capacity of this virus to reach higher titers in the brains of experimentally infected suckling mice. We also found that the Gc glycoprotein, encoded by the M segment of SBVp32, facilitates host cell protein shutoff in vitro. Interestingly, while the M segment of SBVp32 is a virulence factor, we found that the S segment of the same virus confers by itself an attenuated phenotype to wild type SBV as has lost the ability to block the innate immune system of the host. Single mutations present in the Gc glycoprotein of SBVp32 are sufficient to compensate both the attenuated phenotype of the SBVp32 S segment and the attenuated phenotype of NSs deletion mutants. Our data also indicate that the SBVp32 M segment does not act as an IFN antagonist. Therefore SBV mutants can retain pathogenicity even when they are unable to fully control the production of IFN by the infected cells. Overall, this study suggests that the viral glycoprotein of orthobunyaviruses can compensate, at least in part, the function of NSs. In addition, we also provide evidence that the induction of total cellular protein shutoff by SBV is determined by multiple viral proteins while the ability to control the production of IFN maps to the NSs protein. Importance The identification of viral determinants of pathogenesis is key to the development of prophylactic and interventions measures. In this study we found that the bunyavirus Gc glycoprotein is a virulence factor. Importantly, we show that mutations in the Gc glycoprotein can restore pathogenicity of attenuated mutants resulting from deletions or mutations in the non-structural protein NSs. Our findings highlight the fact that careful consideration should be taken when designing live attenuated vaccines based on deletions of non-structural proteins since single mutations in the viral glycoproteins appear to revert attenuated mutants to virulent phenotypes

Virus and host factors affecting the clinical outcome of bluetongue virus infection

Bluetongue is a major infectious disease of ruminants caused by bluetongue virus (BTV), an arbovirus transmitted by Culicoides. Here, we assessed virus and host factors influencing the clinical outcome of BTV infection using a single experimental framework. We investigated how mammalian host species, breed, age, BTV serotypes, and strains within a serotype, affect the clinical course of bluetongue. Results obtained indicate that in small ruminants there is a marked difference in the susceptibility to clinical disease induced by BTV at the host species level, but less so at the breed level. No major differences in virulence were found between divergent serotypes (BTV-8 and BTV-2). However, we observed striking differences in virulence between closely related strains of the same serotype collected towards the beginning and the end of the European BTV-8 outbreak. As observed previously, differences in disease severity were also observed when animals were infected with either blood from a BTV-infected animal or from the same virus isolated in cell culture. Interestingly, with the exception of two silent mutations, full viral genome sequencing showed identical consensus sequences of the virus before and after cell culture isolation. However, deep sequencing analysis revealed a marked decrease in the genetic diversity of the viral population after passaging in mammalian cells. In contrast, passaging in Culicoides cells increased the overall number of low frequency variants compared to virus never passaged in cell culture. Thus, Culicoides might be a source of new viral variants and viral population diversity can be another factor influencing BTV virulence

Molecular determinants of bluetongue virus virulence

Bluetongue virus (BTV) is an arbovirus and the cause of “bluetongue”, a major infectious disease of ruminants. Whilst the BTV structure and replication strategies are well elucidated, less is known on the genetic variability of BTV and the molecular determinants affecting virus-host interactions. In order to investigate the determinants of BTV virulence, in this study, we compared the phenotype and genotype of a highly virulent strain of BTV-8 isolated in the Netherlands a passaged minimally in tissue culture (BTV8L), with a strain passaged extensively in tissue culture (BTV8H). BTV8L was shown to be highly pathogenic in sheep and in a mouse model of bluetongue, while BTV8H was attenuated in both hosts. Full genome sequencing revealed differences in 16 amino acid residues between these two strains. Using reverse genetics, we rescued both viruses, in order two further dissect their biological features. Rescued viruses retained the phenotype of the parental viruses in vivo and in vitro. Reassortants between BTV8L and BTV8H showed that mutations in several segments contributed to attenuation of the high passage virus. The major determinants of BTV8 virulence in IFNAR-/- mice were shown to be located in segments 1, 2, 6 and 10. In vitro studies of selected reassortants showed that through extensive passage in tissue culture BTV8H acquired increased affinity for glycosaminoglycans. This property was conferred by mutations in segment 2 and resulted in increased yields of the virus in vitro and attenuation in vivo. Additionally, BTV8H was unable to replicate in IFN competent primary sheep endothelial cells. Our data showed that multiple segments were involved in decreased efficiency of BTV8H replication in cells in an IFN-induced antiviral state. Moreover, we examined changes in viral population diversity that occured after BTV-8 isolation in insect cells (Culicoides, KC) and after passage in mammalian cells and linked decreased diversity with BTV virulence in vivo. We found, that in general, the number of genetic variants was higher in BTV-8 before cell passaging, or after one passage in KC cells, compared to the number observed after even a single passage in BHK-21 cells. The highest total number of variants was found in virus passaged in KC cells, which suggests that insect vector might serve as an amplifier of quasispecies diversity of BTV. Together, these findings suggest that the virulence of BTV is a multifactorial phenomenon involving many aspects of virus-host interactions and it is not only affected by changes in the viral proteins selected at the consensus level, but also by the genetic variability of the population as a whole

Parafie jako centra aktywności społecznej dla seniorów i wsparcia osób u kresu życia w środowisku domowym. Dobre praktyki i przykłady prowadzonych działań.

The need for mobilization of local communities to the aging population and the increase in the number of subsidiaries and dependent in their homes, raises the question about the role of volunteering and its associated social forces and its possible impact on improving care at the end of life in Poland. Is the successful operation of palliative and hospice care and change of public awareness to the area of social action can be deployed to sensitize society to the needs of all persons at the end of life? What conditions must be met in order to difficult in many respects the situation seriously ill patients and informal caregivers, especially in home care, would support the involvement of volunteers from the local communities? Is it possible the involvement of the parish, which is one of the basic elements of local communities, to create a strategy for integrated care at end of life with the participation of volunteer work? These questions will try to answer this article, describing the process of integration of the local environment by supporting caregivers and the development of voluntary work, then needs care to seniors in local communities and a project to support them through a system of volunteer centers created by catholic parishes in Poland.Potrzeba aktywizacji środowisk lokalnych wobec starzenia się społeczeństwa i zwiększania się liczby osób zależnych i niesamodzielnych w domach, skłania do postawienia pytania o rolę wolontariatu i związanych z nim sił społecznych oraz jego możliwy wpływ na poprawę opieki nad osobami u kresu życia w Polsce. Czy udane działania opieki paliatywno-hospicyjnej i zmiana świadomości społecznej odnośnie do tego obszaru działań społecznych dadzą się wykorzystać do uwrażliwienia społeczeństwa na potrzeby wszystkich osób u kresu życia? Jakie warunki muszą zostać spełnione, by trudną pod wieloma względami sytuację ciężko chorych i opiekunów nieformalnych, zwłaszcza w opiece domowej, mogło wesprzeć zaangażowanie wolontariuszy ze środowisk lokalnych? Czy możliwe jest zaangażowanie parafii, będącej jednym z podstawowych elementów społeczności lokalnych, do stworzenia strategii zintegrowanej opieki nad osobami u kresu życia z udziałem wolontariatu? Na te pytania będzie się starał odpowiedzieć ten artykuł, opisujący proces integracji środowiska lokalnego poprzez wspieranie opieki paliatywno-hospicyjnej i rozwój wolontariatu, następnie potrzeby opiekuńcze wobec seniorów w społecznościach lokalnych oraz projekt ich wspierania poprzez system wolontariatu i centrów senioralnych przy parafiach

Schmallenberg virus pathogenesis, tropism and interaction with the innate immune system of the host

Schmallenberg virus (SBV) is an emerging orthobunyavirus of ruminants associated with outbreaks of congenital malformations in aborted and stillborn animals. Since its discovery in November 2011, SBV has spread very rapidly to many European countries. Here, we developed molecular and serological tools, and an experimental in vivo model as a platform to study SBV pathogenesis, tropism and virus-host cell interactions. Using a synthetic biology approach, we developed a reverse genetics system for the rapid rescue and genetic manipulation of SBV. We showed that SBV has a wide tropism in cell culture and “synthetic” SBV replicates in vitro as efficiently as wild type virus. We developed an experimental mouse model to study SBV infection and showed that this virus replicates abundantly in neurons where it causes cerebral malacia and vacuolation of the cerebral cortex. These virus-induced acute lesions are useful in understanding the progression from vacuolation to porencephaly and extensive tissue destruction, often observed in aborted lambs and calves in naturally occurring Schmallenberg cases. Indeed, we detected high levels of SBV antigens in the neurons of the gray matter of brain and spinal cord of naturally affected lambs and calves, suggesting that muscular hypoplasia observed in SBV-infected lambs is mostly secondary to central nervous system damage. Finally, we investigated the molecular determinants of SBV virulence. Interestingly, we found a biological SBV clone that after passage in cell culture displays increased virulence in mice. We also found that a SBV deletion mutant of the non-structural NSs protein (SBVΔNSs) is less virulent in mice than wild type SBV. Attenuation of SBV virulence depends on the inability of SBVΔNSs to block IFN synthesis in virus infected cells. In conclusion, this work provides a useful experimental framework to study the biology and pathogenesis of SBV

Znaczenie prognostyczne ultrasonograficznego rozpoznania krwiaka podkosmówkowego dla przebiegu ciąży

Objectives: the aim of this retrospective study was to assess if a first trimester subchorionic hematoma (SCH) influences the pregnancy outcome and whether pv bleeding can be a prognostic factor for the pregnancy course. Material and methods: the study included 185 pregnant women hospitalized due to symptoms of a threatening miscarriage. Patients were divided into 2 groups: 119 women with SCH (study group) and 66 patients with normal prenatal scan (control group), further subdivided into cases with and without pv bleeding, irrespectively of the outcome of the ultrasound scan. Obstetric and neonatal data were analyzed. Results: 1. A pregnancy complicated by SCH is more often associated with a poor outcome – 23.78% of the study group patients had a miscarriage vs. 1.62% of the controls. 2. Pregnancy with SCH is more likely to be lost before 9 weeks of gestation. 3. The ‘N” ratio, that expresses the maximal length of the hematoma to the maximal length of the fetus, equal to 2.5 or more, is associated with a risk of miscarriage. 4. The surface area of SCH equal to 280mm2 or more is more likely to reveal with vaginal bleeding. 5. Vaginal bleeding can be a prognostic factor for the mode of delivery – a higher rate of the Cesarean section is observed in patients with pv bleeding. 6. SCH is a complication that occurs in older women, with the limit of 30 years of age. 7. In this study there were no significant correlations between subchorionic hematoma or pv bleeding and PTL, IUGR, PIH, abnormal volume of the amniotic fluid, parity and order of gestation and delivery. Conclusions: subchorionic hematoma can be associated with poor pregnancy outcome and the ”N” index may be a useful predictor of the further course of a pregnancy. Pv bleeding may be a prognostic factor for the delivery mode.Cel pracy: Celem pracy była retrospektywna analiza przebiegu ciąż powikłanych obecnością krwiaka podkosmówkowego (subchorionic hematoma – SCH) zdiagnozowanego w I trymestrze ciąży. Pytaniem badawczym była ocena czy obecność SCH ma wpływ na dalszy przebieg ciąży oraz czy krwawienie z dróg rodnych, mogące towarzyszyć tej patologii, jest dla niej czynnikiem prognostycznym. Materiał i metody: Badanie objęło grupę 185 ciężarnych hospitalizowanych w I trymestrze ciąży z powodu objawów poronienia zagrażającego. Pacjentki podzielono na 2 grupy: grupa badana (119 kobiet) z rozpoznanym ultrasonograficznie SCH i grupa kontrolna (66 pacjentek) z prawidłowym wynikiem badania USG. W obrębie tych grup wyodrębniono kolejne dwie, które podzielono ze względu na obecność krwawienia z dróg rodnych bądź jego brak, niezależnie od wyniku badania USG. Wyniki: Obecność SCH wpływa na rozwój ciąży, prowadząc częściej do jej straty. Poronienia w grupie badanej stanowiły 23,78% vs 1,62% w grupie kontrolnej. Ciąża powikłana obecnością SCH do 9. tygodnia, częściej ulega poronieniu, w porównaniu do ciąży starszej. Współczynnik ‘N’, przedstawiający wielkość krwiaka względem płodu, ≥2,5 jest skorelowany z częstszym występowaniem poronienia. Powierzchnia SCH ≥280mm2 daje objawy poronienia zagrażającego w postaci krwawienia per vaginam. Ciężarne z krwawieniem z dróg rodnych we wczesnej ciąży częściej rodzą drogą cięcia cesarskiego. SCH częściej dotyczy kobiet po 30 r.ż. Wnioski: 1. Obraz SCH rozpoznany w USG w I trymestrze ciąży jest czynnikiem ryzyka jej dalszego rozwoju, istotnie częściej prowadzącym do jej straty, niezależnie od obecności objawów klinicznych w postaci krwawień z dróg rodnych, bądź ich braku. 2. Współczynnik ‘N’, przedstawiający wielkość krwiaka względem rozwijającego się płodu, jest pomocnym narzędziem, pozwalającym na prognozowanie dalszego przebiegu ciąży. 3. Krwawienie z dróg rodnych we wczesnej ciąży może być czynnikiem prognostycznym sposobu jej rozwiązania

Zespołowa opieka duszpasterska jako forma opieki duchowej w ochronie zdrowia i pomocy społecznej

In addition to the traditional model of individual pastoral assistance chaplains exist in many countries, pastoral care team with the participation of clergy and laity, open to ecumenical and interreligijną cooperation for the sake of spiritual and religious needs of residents. Also in Poland, pastoral care teams should take care not to, with respect for religious and cultural diversity. In order to do this effectively we need new tools to study the spiritual needs, in addition to the traditional tools of pastoral care and sacramental care. There are numerous tools for studying religion, but the lack of working tools study the spiritual needs of the residents. Such an assumption meets the qualitative questionnaire FICA by prof. Christina M. Puchalski. His adaptation into Polish can help you turn a practical spiritual dimension in the care process.Obok tradycyjnego modelu indywidualnej pomocy duszpasterskiej kapelanów istnieje w wielu krajach zespołowa opieka duszpasterska z udziałem duchownych i świeckich, otwarta na ekumeniczną i interreligijną współpracę w trosce o potrzeby duchowo-religijne podopiecznych. Również w Polsce zespoły opieki duszpasterskiej powinny zatroszczyć się o nie, z szacunkiem dla różnorodności religijnych i kulturowych. By móc to skutecznie czynić potrzebne są nowe narzędzia do badania potrzeb duchowych, obok tradycyjnych narzędzi duszpasterskiej troski i sakramentalnej opieki. Istnieją liczne narzędzia badania religijności, ale brak wypracowanych narzędzi badania duchowych potrzeb podopiecznych. Takie założenie spełnia jakościowy kwestionariusz FICA autorstwa prof. Christiny M. Puchalski. Jego adaptacja na język polski może pomóc w praktycznym włączaniu wymiaru duchowego w proces opieki

Role of psychological and emotional factors in cancer related fatigue (CRF) syndrome in advanced NSCLC patients undergoing palliative chemotherapy

Background: Cancer-related fatigue syndrome (CRF) is observed in most cancer patients, especially in those with advanced disease. Pathomechanism of CRF is complex and not fully understood. The aim of the study was to determine the role of psychological and emotional factors in CRF in non small-cell lung cancer (NSCLC) patients at advanced stage of the disease undergoing chemotherapy. Material and methods: 76 patients with locally advanced or metastatic NSCLC, undergoing palliative chemotherapy participated in the study. Prior to, during and after the treatment, the patients were asked to complete psychological questionnaires, like HADS, CERQ and daily diary constructed for the purpose of this study. Results: Coping strategies like concentrating on other positive aspects and acceptance are factors, which positively, while catastrophising, anxiety and depression – negatively, characterise the level of perceived cancer related fatigue. Conclusions: Results show that some coping and emotional strategies as well as psychological features may constitute predictors for CRF. It may suggest that psychological features can influence the performance status of patients and reduce fatigue perceived by patients. Further research is needed to fully explained the phenomenon. Adv. Pall. Med. 2010; 9, 3: 81–86Background: Cancer-related fatigue syndrome (CRF) is observed in most cancer patients, especially in those with advanced disease. Pathomechanism of CRF is complex and not fully understood. The aim of the study was to determine the role of psychological and emotional factors in CRF in non small-cell lung cancer(NSCLC) patients at advanced stage of the disease undergoing chemotherapy. Material and methods: 76 patients with locally advanced or metastatic NSCLC, undergoing palliative chemotherapy participated in the study. Prior to, during and after the treatment, the patients were asked to complete psychological questionnaires, like HADS, CERQ and daily diary constructed for the purpose of this study. Results: Coping strategies like concentrating on other positive aspects and acceptance are factors, which positively, while catastrophising, anxiety and depression – negatively, characterise the level of perceived cancer related fatigue. Conclusions: Results show that some coping and emotional strategies as well as psychological features may constitute predictors for CRF. It may suggest that psychological features can influence the performance status of patients and reduce fatigue perceived by patients. Further research is needed to fully explained the phenomenon. Adv. Pall. Med. 2010; 9, 3: 81–8

Epidermal growth factor receptor (EGFR) inhibition on non-small-cell lung cancer treatment

Niedrobnokomórkowy rak płuca (NDRP) nadal stanowi istotny problem kliniczny i społeczny. W ostatnim okresie notuje się stały wzrost zachorowań w populacji kobiet i mężczyzn. Wyniki leczenia chirurgicznego, radioterapii i chemioterapii osiągnęły plateau. Perspektywy farmakologicznego leczenia NDRP warunkuje poznanie mechanizmów molekularnych odpowiedzialnych za rozwój nowotworu oraz powstawanie oporności. Wprowadzenie leków ukierunkowanych molekularnie stanowi istotny postęp w leczeniu NDRP. Leczenie ukierunkowane obejmuje obecnie zastosowanie leków antyangiogennych oraz monoklonalnych przeciwciał i drobnocząsteczkowych inhibitorów kinazy tyrozynowej. Ogromną rolę odgrywa identyfikacja predykcyjnych czynników pozwalająca na optymalizację leczenia. Onkol. Prak. Klin. 2011; 7, 4: 177–182Non small cell lung cancer (NSCLC) remains a significant clinical and social problem. In the last period there has been a steady increase a incidence in women and men. Results of surgical treatment, radiotherapy and chemotherapy have reached a plateau. Perspectives for pharmacological treatment of NSCLC determines understanding the molecular mechanisms responsible for tumor growth and resistance. The introduction of molecularly targeted drugs is an important step in the treatment of NSCLC. Targeted treatment includes the use of monoclonal antibodys, anti-angiogenic treatment and small molecule tyrosine kinase inhibitors. Identity of predictive factors plays a huge role for targeting treatment optimization. Onkol. Prak. Klin. 2011; 7, 4: 177–18

Species biodiversity of microscopic fungi of common reed (Phragmites australis (Cav.) Trin. ex Steud.) in the rush communities of Lake Glinno

The studies were carried out on common reed (&lt;i&gt;Phragmites australis&lt;/i&gt;) growing on the shores of Lake Glinno and forming rush communities of the alliance &lt;i&gt;Phragmition&lt;/i&gt;. 10 plants with disease symptoms were gathered from each of five sites. The isolation and marking of pathogens were performed twice from fragments of leaf and blade tissues with disease symptoms. First, directly after collecting the plants incubated in sterile humid chambers and microorganism cultures on CDA and PDA medium, and then phytopathogen and saprotroph species occurring on dried green material were identified for 2-4 months. The occurrence of 31 species of microscopic fungi overall was observed on the leaves, blades and inflorescences of &lt;i&gt;P. australis&lt;/i&gt;, including 2 mycelia of &lt;i&gt;Mycelia sterilia&lt;/i&gt;. The most frequently occurring species, present at all sites of &lt;i&gt;Phragmites australis&lt;/i&gt; are: &lt;i&gt;Alternaria alternata&lt;/i&gt;, &lt;i&gt;Cladosporium cladosporioides&lt;/i&gt;, &lt;i&gt;C. herbarum&lt;/i&gt;, &lt;i&gt;Doratomyces stemonitis&lt;/i&gt; and &lt;i&gt;Puccinia phragmitis&lt;/i&gt;, &lt;i&gt;P. magnussiana&lt;/i&gt; and two mycelia of &lt;i&gt;Mycelia sterilia&lt;/i&gt;. The most common species occurring on blades and inside them are: &lt;i&gt;Acremoniella atra&lt;/i&gt;, &lt;i&gt;Acremonium alternatum&lt;/i&gt; and &lt;i&gt;Fusarium sambucinum&lt;/i&gt;. Sporadically, &lt;i&gt;Ustilago grandis&lt;/i&gt; was also observed inside blades. Tiny necrotic stains on leaves and blades were caused by the presence of three species of the genus Leptoshaeria: &lt;i&gt;L. culmifraga&lt;/i&gt;, &lt;i&gt;L. eustoma&lt;/i&gt; and &lt;i&gt;L. fuckelli&lt;/i&gt;. The occurrence of the sclerote of &lt;i&gt;Claviceps microcephala&lt;/i&gt; was found in inflorescences.</jats:p