Despite the similar name, PCMV is more closely related with human herpesvirus-6 (HHV-6) and HHV-7, which are also Roseoloviruses, but not so closely with the human cytomegalovirus (HCMV), also called HHV-5 [ 31 , 35 , 36 ]. HCMV is the cause of disease in the human fetus, the allograft recipient, AIDS patient, those admitted to inten- sive care units, and in the elderly [ 37 ]. HCMV is the single most important infectious agent affecting recipients of organ transplants, with at least two-thirds of these pa- tients having CMV infection after transplantation [ 38 ]. During solid organ transplantation, seropositive donors frequently transmit HCMV to seronegative recipients (pri- mary infection) [ 38 , 39 ]. Primary infection has the greatest clinical impact. In addition, during allotransplantation HCMV may be re-activated bythe allo-immune responses and bythe immunosuppression in the HCMV-positive re- cipient [ 40 ]. Finally, superinfections have been described
Viral infection: Furthermore pigs harbor several infectious viruses which have a pathogenic potential for human. These include for example the Nipah virus  Menangle virus , and Tioman virus , hepatitis E virus (HEV) , porcinecytomegalovirus (PCMV) which is able to infect human fibroblasts in vitro , porcine gammalymphotropic herpesvirus (PLHV)  and the influenza virus which can cross the species barrier and infect humans . Analyses ofthe H1N1 1918/1919 influenza virus, which killed at least 40 million people, revealed that this strain might have originated from the swine influenza virus . The vast majority of these viruses can be eliminated from thepig herd using sensitive detection assays for screening, by maintaining pigs under SPF conditions and using specialized animal husbandry. Although no transmission of potential infectious viruses was detected during the last decade in humans exposed to live porcine cells , some viruses are still pose a relevant risk for xenotransplantation especially the viruses whose DNA is integrated into the germline of pigs, such as theporcine endogenous retroviruses (PERVs) which can infect human cells in vitro and are widely distributed in the genome of pigs and thus cannot yet be eradicated .
Received: 5 June 2019; Accepted: 9 July 2019; Published: 16 July 2019 Abstract: Porcine circovirus 3 (PCV3) is a newly described member ofthe virus family Circoviridae. PCV3 is highly distributed among pigs and wild boars worldwide. A sudden introduction of PCV3 was recently observed in a herd of triple genetically modified pigs generated for xenotransplantation. These animals were used as donor pigs for orthotopic heart transplantation into baboons. In four cases, PCV3-positive hearts were transplanted, and transmission of PCV3 to the recipient was observed. PCV3 was found in all organs ofthe recipient baboons and a higher virus load was found in animals with a longer survivaltimeofthe transplant, indicating replication ofthe virus. This is the first report showing trans-species transmission of PCV3 to baboons by transplantation of a heart from a PCV3-positive donor pig. Sequence analysis showed that PCV3a and PCV3b were present in the infected pigs and were transmitted. Experiments to infect human 293 cells with PCV3 failed.
Abstract: Porcinecytomegalovirus (PCMV) infection is widely prevalent among pigs, and PCMV is one ofthe viruses which may be transmitted during xenotransplantation using pig cells, tissues, or organs. While human cytomegalovirus (HCMV) is a major risk factor for allotransplantation, it is still unclear whether PCMV is able to infect human cells or pose a risk for xenotransplantation. Previously, it was shown that transmission of PCMV after pig kidney to non-human primate transplantations resulted in a significantly reduced survivaltimeofthe transplanted organ. To detect PCMV, PCR-based and immunological methods were used. Screening of pigs by Western blot analyses using recombinant viral proteins revealed up to 100% ofthe tested animals to be infected. When the same method was applied to screen human sera for PCMV-reactive antibodies, positive Western blot results were obtained in butchers and workers in the meat industry as well as in normal blood donors. To exclude an infection of humans with PCMV, the sera were further investigated. PCMV is closely related to human herpesvirus-6 (HHV-6) and human herpesvirus-7 (HHV-7), and a sequence alignment of glycoprotein B suggests that the antibodies may cross-react with identical epitope sequences. HCMV is not related with PCMV, and no correlation between antibody reactivity against PCMV and HCMV was detected. These data indicate that antibodies against PCMV found in humans are cross-reactive antibodies against HHV-6.
Xenotransplantation has been proposed as a solution to the shortage of suitable human donors for transplantation and pigs are currently favoured as donor animals. However, xe- notransplantation may be associated with the transmission of zoonotic microorganisms. Whereas most porcine microorganisms representing a risk for the human recipient may be eliminated by designated pathogen free breeding, multiple copies ofporcine endogenous retroviruses (PERVs) are integrated in the genome of all pigs and cannot be eliminated this way. PERVs are released as infectious particles and infect human cells. The zinc finger nu- clease (ZFN) technology allows knocking out specifically cellular genes, however it was not yet used to eliminate multiple integrated proviral sequences with a strong conservation in the target sequence. To reduce the risk of horizontal PERV transmission and to knock out as many as possible proviruses, for the first timethe powerful tool ofthe ZFN technology was used. ZFN were designed to bind specifically to sequences conserved in all known rep- lication-competent proviruses. Expression and transport ofthe ZFN into the nucleus was shown by Western blot analysis, co-localisation analysis, PLA and FRET. Survivalof trans- fected cells was analysed using fluorescent ZFN and cell counting. After transfection a strong expression ofthe ZFN proteins and a co-localisation ofthe expressed ZFN proteins were shown. However, expression ofthe ZFN was found to be extremely toxic for the trans- fected cells. The induced cytotoxicity was likely due to the specific cutting ofthe high copy number ofthe PERV proviruses, which is also commonly observed when ZFN with low specificity cleave numerous off-target sites in a genome. This is the first attempt to knock out multiple, nearly identical, genes in a cellular genome using ZFN. The attempt failed, and other strategies should be used to prevent PERV transmission.
Since pig 5154 was PCMV-positive in a few organs, but negative when the blood was tested using the same real-time PCR, an analysis was performed as to whether testing for PCMV in the blood can be improved by incubating the isolated PBMCs from the animal under study for five to seven days in culture medium. Freshly isolated PBMCs from nine pigs were incubated and tested before and after incubation (Figure 3). These animals were DanAvl basic hybrid sows; they were not genetically modified, and they had been derived from a SPF facility in Fehmarn, Germany. Twenty-eight days before this experiment, blood samples from these animals tested negative for PCMV using PCR. After cultivation ofthepig PBMCs, a significant increase in the amount of detected virus was observed in two of nine cases (pigs 91106, 91111), and a moderate increase was observed in three of nine cases. In four cases, no virus was detected; presumably, these animals were truly negative, at least in the blood samples and using our detection method. Stimulating the PBMCs with the mitogen PHA did not increase the virus replication. These data indicate that PCMV was replicating during cultivation, and that this approach increases the efficacy of virus screening in the blood, which may be a great advantage when testing pigs. This is a very important finding. Without incubation, all nine animals would have been declared PCMV-negative, but using this assay, five animals were found PCMV-positive.
Table 2. Age and boulder density for craters for which an age estimate is available. Density is defined as the number of boulders larger than 3 pixels divided by crater equivalent area. The densities in brackets are underestimates, as the associated craters were largely in the shadow in lamo images.
= 0.49兲, further reinforce the suggestion that the bulk sound speed ofthe rendered porcine fat tested here should be lower than the linear-fit value of c 0 = 1.58 mm / s.
The main mechanism proposed to explain curvature ofthe U s -u p Hugoniot at lower particle velocities involves two stages: 共1兲 primary reduction in the spaces between the poly- meric chains, due to the existence of weak van der Waals forces, before; 共2兲 secondary compression ofthe backbone carbon chain. Although this mechanism is arguably appli- cable to polymers in general, Carter et al. 16 discussed the existence of phase changes within certain polymeric materi- als at low pressures and how these could also account for such observations. Further, other theories describing this be- havior in polymers also exist, such as the elastic-plastic flow model, 36 which predicts extreme curvature ofthe Hugoniot at low particle velocities. Consequently, the inclusion ofthe data-point corresponding to an impact velocity of 39 m/s, suggesting slight-curvature in the Hugoniot ofthe adipose material at low particle velocities, supports evidence that the material possesses structural similarity to simple polymers.
often omitted from boulder (and crater) counting papers. Several unfortunate practices associated with the cumulative distribution have become established in the literature. The first is binning. The Crater Analysis Techniques Working Group (1979) wrote that the “collection, manipulation, and display of unbinned data is more time consuming than for binned data.” This may have been true in 1979 but is no longer the case. Binning of a cumulative distribution is not merely unnecessary but represents a loss of information. Also, bins that have the same value as their neighbor on the right are often not displayed. Their omission skews the appearance ofthe distribution and affects how we perceive the goodness of fit of a model curve. Another statistically suspect practice is the association of Poisson (square root) error bars with the bins. Such error bars are valid for single bins when considered in isolation, but in a plot ofthe cumulative distribution they do not serve their usual purpose of indicating the uncertainty ofthe data: Regardless ofthe size of its bar, the number in a bin can neither be lower than its neighbor on the right nor higher than its neighbor on the left. Therefore, we do not bin the cumulative distribution in this paper. The differential distribution is always binned in practice, where the bin width can be chosen as constant on a linear or logarithmic scale. Poisson error bars associated with the bins are statistically meaningful and will give the correct impression of how well the data agree with a particular model curve. Empty bins, however, present a problem. SFD plots are invariably shown on a log-log scale, and empty bins cannot be displayed, which skews the appearance ofthe distribution. Moreover, empty bins cannot be included when fitting models (such as a power law) to the logarithm ofthe data, introducing bias.
Z. N aturforsch. 4 8 c , 451 - 4 5 6 (1993); received December 23, 1992/February 24, 1993 M icrobial, Asymmetric, R eduction, Marine, M icroorganism
Three strains o f bacteria reducing (trifluoroacetyl)ferrocene (3) to optically pure (R )-2,2,2- trifluoro-l-hydroxyethylferrocene (4) and one bacterial strain reducing 3 to (5^)-4 o f moderate optical purity were isolated from sea-water collected at the coastal areas in Ibaraki prefecture o f Japan. The former three strains were identified as M icrococcus lylae, M icrococcus luteus, and D eleya marina and the latter as Bacillus licheniformis. These strains also asymmetrically reduced som e other synthetic ketones, e.g., 2,2,2-trifluoroacetophenone (7) and phenyl tri- m ethylsilyl ketone (9). Further screening o f microorganism s capable o f reducing 3 was done with bacteria isolated from sea-water o f the deep sea (O kinawa trough, Japan trench, and M ariana trough) and o f the pelagic areas (Indian Ocean and South China Sea). M ost o f these marine strains preferentially reduced 3 to (/?)-4 similar to the coastal strains, but the frequency o f finding very highly enantioselective strains (i.e., those form ing 4 o f > 9 0 % e.e.) was remark ably high in several sites o f the deep sea and pelagic areas as com pared with the coast and terrestrial environm ent.
Peritoneal carcinosis can be a result of intraperitoneal tumor cell spread after surgical treatment of colonic can- cer. Tumor cell attachment occurs through blood-or lym- phatic vessels or by accidentally opening the colonic specimen . Serosal invasion ofthe primary tumor leads in up to 50% of patients to intraperitoneal metastases . However, even stage 1 and stage 2 of colonic cancer, gas- tric cancer, uterine cancer or pancreatic cancer may cause peritoneal dissemination . In gastrointestinal cancer the detection of free intraperitoneal tumor cells, serves as an independent prognostic factor . Free floating intra- peritoneal tumor cells may attach to, degrade and migrate through the extracellular matrix (ECM) . Particularly if the peritoneum is damaged and the etracellular matrix is exposed tumor cell adhesion accumulates . In former studies we found a widespread peritoneal carcinosis, with tumor cell adhesion in most peritoneal aeras after intraab- dominal instillation of tumor cells (Cell line: DHD/K12/ TRb). We could demonstrate that tumor cells predomi- nantly adhere to injured peritoneal areas. However, the goal ofthe underlying study was not locoregional recur- rence but the effect of phospholipids in the complete peri- toneal cavity. In addition preliminary studies could show that a phospholipid emulsion is able to significantly reduce intraperitoneal tumor cell adhesion [3,6,7]. Phospholipids are natural constituents of peritoneal fluid secreted by mesothelial cells. These polar phosphoric acid di-esters are capable to form a lubricant layer on the peri- toneal surface, which is of paramount importance to pre- vent adhesion [19,20]. Additionally, treatment with phospholipids (e.g. gangliosides) affect integrin function, causing reduced cell motility and adhesion capability after exogenous addition of phospholipids [8-10]. Further- more other adhesion-preventing substances are known. For example Jeekel et al. evaluated the effects of intra- abdominal treament with Icodextrin, a glucose polymer solution, in a coloncarcinoma CC531 rat model . As we could demonstrate the positive effect of phospholi- pids in low dosage in former studies, the aim ofthe under- lying experiment was to compare the influence of rising phospholipid concentrations on the one hand and differ- ent tumor cell concentrations on the other hand with spe- cial emphasis on possible side effects.
(TEA; 99.5%), and tetrahydrofurane (THF, anhydrous, ≥99.9%, inhibitor-free) were purchased from Sigma-Aldrich (St. Louis, USA). Lithium aluminum hydride (LAH) (1.0 M in THF), (+)-PHNO standard, and precursor (+)-HNO hydrochloride (GMP and non-GMP) were obtained from ABX (Advanced Biochemical Compounds, Radeberg, Ger- many). All reagents were used without further puriﬁcation. Sterile water and 0.9% saline solution were purchased from B. Braun (Melsungen, Germany). Sterile phosphate-buﬀered saline solution (0.021 M phosphate buﬀer, 0.188 M·NaCl, pH 7.7) was obtained from the Vienna General Hospital’s Pharmacy (Vienna, Austria). The loop for the Grignard reactions is made of a 90 cm polyethylene (PE) tubing (ﬁne bore polythene tubing REF 800/100/280; ID: 0.86 mm OD: 1.52 mm, Smiths Medical International Ltd.; Kent, UK). Ascarite II (20–30 mesh) was purchased from Thomas Scientiﬁc (Swedesboro, USA). Removal of acetonitrile was performed by using solid-phase extraction (SPE) cartridges (SepPak C18-plus) from Waters (Waters Cooperation; Milford, MA, USA). [ 11 C]CO
The probe was generated by PCR using the primers Apa2-for and Apa2-rev (section 2.1.8) on pSM3fr-FRT as template. The PCR product was purified by GFX PCR DNA and Gel Band Purification Kit (section 188.8.131.52). M210B4 cells were infected with MOI 0.1 to 0.5 and total cellular DNA was isolated 48 hpi. From these 1 µg DNA of infected cells was digested by 40 units of ApaLI in a final volume of 40 µl overnight at 37 ◦ C. The DNA was then applied to agarose gel electrophoresis at 80 V for 24 h. The gel size was reduced to minimal size for detection of required bands and size marker was documented by photogra- phy ofthe gel next to a ruler. To mobilise the DNA in the gel, it was incubated in 250 mM HCl twice for 10 min and subsequently for 45 min in denaturation buffer. Afterwards the gel was incubated for 30 min, followed by additional 10 min, in neutralisation buffer. The DNA was then transferred by capillary force to a membrane (Amersham Hybond TM - N + , GE Healthcare). For this, blotting paper (Macherey-Nagel) in gel width was placed in an agarose gel electrophoresis chamber with both ends dunked in 20x SSC buffer. The gel was placed onto the paper and the membrane in gel size on top ofthe gel. The capillary force was established by 2 gel size blotting papers soaked in 20x SSC and 2 additional dry ones, followed by a tower of tissue papers. This was fixed by weight and balanced. Blotting was performed overnight.
Despite this considerable public health burden, few women are aware of congenital CMV infection [ 7 – 9 ]. Educating women about CMV transmission and preventive hygiene behaviour can significantly reduce primary CMV infections during pregnancy and thereby congenital CMV infections [ 10 – 14 ]. A vaccine would be necessary to significantly and permanently reduce congenital (and other) CMV infections. To date, there is no licensed vaccine available that protects against CMV. However, several vaccine candidates are currently being tested in clinical trials [ 15 – 17 ]. A vaccine against CMV was classified as a top priority by "The National Vaccine Advisory Committee" in the US in 2004, based on the estimation that the disease bur- den of congenital CMV infection is as high as the disease burden due to congenital rubella before the introduction of rubella vaccinations [ 18 , 19 ]. Representative epidemiological data on the CMV susceptibility ofthe population are essential for decision making in the fields of public health and primary prevention through immunization. Since there has been no pop- ulation-based CMV-specific Ig seroprevalence data available for German adults, the aims of this study were to estimate CMV seroprevalence in the adult population in Germany and to identify socio-demographic and lifestyle factors that are potentially associated with CMV seropositivity.
A first study has been conducted for the evaluation ofthe performance enhancement of a remote satellites constellation served by a small or micro satellite constellation for commands delivery and data collection. Main performance figure of merit considered is the response time. Other significative figures of merit are the information age and the total number of satisfied user requests. Numerical simulations have been performed for a SAR satellite constellation in LEO serviced by a small-satellite constellation in different geometrical configurations and altitudes. The simulation scenario has been kept at a lowest possible level of complexity in order to evaluate only the potentialities ofthe physical system and no scheduling optimization method but a FIFO approach has been used for the building ofthe operations schedule. The simulation results are promising, though they cannot be considered conclusive. As only telecommands bridging capability has been exploited in the simulations, a future analysis should aim to evaluate the advantages afforded bythe servicing satellites when exploited in their full capabilities. A simulation scenario should also be built for a more effective evaluation ofthe results. A large number of simulations has to be conducted considering different Earth observation systems and servicing satellite constellation configurations in order to have some statistical results. Though a cost analysis has not been performed, it shows up that a C/S constellation can be convenient if the complexity ofthe satellites can be kept at a minimum level and if the system can be used as a service for several Earth observation missions.
wanted to examine the effect in case of lower tumor cell concentrations.
In experiment A we could demonstrate a significant reduc- tion of peritoneal dissemination as measured by all eval- uation methods after treatment with 6%- and 9%- phospholipids. In former studies -using the same tumor cell concentrations but lower phospholipid concentra- tions (1,5% und 3%)- we found a reductionof peritoneal dissemination, too. However, the difference between con- trol-group and treatment-group was not as distinctive as in this case . In experiment B we diversified the number of tumor cells to mimic the clinical situation, that during the resection of a colon cancer only very few tumor cells were released.
w aves (I, I ', I " , II), using a m ercury pool electrode of ap p ro x im ate area 10 cm 2. The reductions were in te rru p te d a t in terv als of tim e to allow polaro- graphic an d UV checks of th e species present. The processes appeared to be independent of both pH a n d co n cen tratio n of th e solution. The reductions corresponding to th e p lateau of wave (I) a t pH < 5 an d of w ave (I” ) betw een pH 6 and 9, involve six electrons per diazoacetone molecule. Thereduction p ro d u ct w as ob tain ed by ex tractio n w ith eth er followed by ev ap o ratio n under reduced pressure. The UV spectrum in w ater of th e resulting colour less liquid is identical w ith th a t of an au th en tic sam ple of 2,5-dim ethylpyrazine (A = 277, 295 (shoulder) nm). The reductions a t th e plateau of w ave (I') correspond to an average n of 2 (between
MCMV M51, M52, M56, M77, M89, M93, and M104. The current model ofthe herpesvirus DNA packaging is based on data mostly acquired bythe study of alpha herpesviruses (Yang and Baines, 2008; Yang et. al., 2008; Yang, Homa and Baines, 2007; Wills et. al., 2006; Yang and Baines, 2006; Beard and Baines, 2004; Taus et. al., 1998). As the core ofthe cleavage/packaging reaction seems to be extremely conserved, a mechanistic analogy with DNA bacteriophage genome packaging has been assumed (Gibson et. al., 2008). Therefore, the terminology ofthe subunits follows that used for bacteriophages. The terminase is recognizing and binding to the genome terminal repeats. Interaction between the portal complex and terminase leads to cleavage ofthe first genome end and the DNA is concomitantly transported through the portal channel to the inside ofthe capsid. When the capsid is full the second genome end is recognized and cut. As a final step, the terminase dissociates from the portal which is then sealed by accessory proteins. The herpesvirus terminase (TER) comprises three conserved units (M89 – TER1; M56 – TER2; M51 – TER3) and docks at the portal vertex. The TER2 recognizes specific DNA sequences of concatemers and performs DNA cleavage, generating linear unit length genomes. TER1 contains a conserved P-loop ATP- ase motif and provide the necessary energy for encapsidation (Wills et. al., 2006; Thurlow et. al., 2005). The M51 homologues (TER3) stabilize the terminase at the docking site and are involved in translocation of cleaved genomic DNA into the capsid’s interior (Beard and Baines, 2004).
A total of 11 patients were identified who received letermovir for the compassionate use for CMV infection. One patient had to be excluded as the exact timeof termination of treatment was not documented. Another patient was excluded as she was already discussed in an earlier case report, which has not yet been published. The other nine patients are discussed below. Table 1 gives an overview ofthe demographic charac- teristics. The median age was 66 years (45–70), and 77.8% ofthe patients was male. Five patients were solid organ trans- plant recipients (55.6%), two developed CMV infection after HSCT (22.2%), one patient suffered from TARFO syndrome (11.1%) and one patient suffered from systemic lupus erythe- matosus (SLE) (11.1%). Six patients experienced asymptom- atic CMV viremia (66.7%), one CMV syndrome (11%), one a probable CMV pneumonia (11%) and one patient with prob- able CMV enteritis (11%) [ 8 ]. Clinical reasoning for compas- sionate use of letermovir was as follows: confirmed antiviral resistance against GCV ( n = 2, 22.2%), virological treatment failure of GCV (n = 1, 11.1%) and HSCT associated with significant CMV viremia (n = 2, 22.2%). In the other four patients, the clinicians opted for letermovir instead of ganci- clovir or foscarnet, to prevent aggravation of coexisting dis- eases. In three ofthe patients severe leukopenia and concom- itant infection (two of them septic) were the reasons for
Leisten Sie Ihren persönlichen Beitrag zur europäischen Energie- sicherheit! Verlassen Sie sich nicht auf die Russen. Energieressour- cen haben und Energieressourcen fördern sind nämlich zweierlei. Russland kauft das Zeug selbst in seinem Nachbarstan. Und bis die Wüstenenergie aus sonnenbespiegelten Wasserkochern, die der Club of Rome propagiert, bei uns ankommt, dürften wir die Grenzen unseres Wachstums sowieso schon ausgereizt haben. Vielleicht können wir von den alten Germanen lernen: Nachdem im zweiten Jahrhundert nach Christus die spanische Überpro- duktion die Olivenölmärkte zusammenbrechen ließ, musste an Lampenöl gespart und auf Wachs und Talg umgestiegen werden. Wir haben es bisher nur bis zur Energiesparlampe geschafft, dafür europaweit. Wärme spendet sie jedoch nicht.