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EN 81-21:2022 - The latest standard for passenger and goods lifts in old structures

This free download details 146 oft-times harrowing stories surrounding avalanches, the lives they claim, survivors and witnesses, along with assessments as to what happened, why it happened, and what could have been done to prevent loss of life and/or property. The authors are never judgmental, their clear-eyed accounts contain a wealth of wisdom that will add to the knowledge-base of any winter backcountry enthusiast: it may be a life-saver!

We predict the bulk thermal conductivity of Lennard-Jones argon and Stillinger-Weber silicon using the Green-Kubo (GK) and direct methods in classical molecular dynamics simulations. While system-size-independent thermal conductivities can be obtained with less than 1000 atoms for both materials using the GK method, the linear extrapolation procedure [Schelling et al., Phys. Rev. B 65, 144306 (2002)] must be applied to direct method results for multiple system sizes. We find that applying the linear extrapolation procedure in a manner consistent with previous researchers can lead to an underprediction of the GK thermal conductivity (e.g., by a factor of 2.5 for Stillinger-Weber silicon at a temperature of 500 K). To understand this discrepancy, we perform lattice dynamics calculations to predict phonon properties and from these, length-dependent thermal conductivities. From these results, we find that the linear extrapolation procedure is only accurate when the minimum system size used in the direct method simulations is comparable to the largest mean-free paths of the phonons that dominate the thermal transport. This condition has not typically been satisfied in previous works. To aid in future studies, we present a simple metric for determining if the system sizes used in direct method simulations are sufficiently large so that the linear extrapolation procedure can accurately predict the bulk thermal conductivity.

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The Seychelles Legal Information Institute (SeyLII) offers free and anonymous access to case law, legislation and secondary legal materials from the Seychelles. Access to the letter of the law supports the rule of law, access to justice and economic development. SeyLII is an independent, self-funded organisation and needs your support.

Next, we tested the in vivo antitumor activity of eCF506 in primary breast cancer models in immunocompetent (FVB wild type) and immunocompromised (CD1 nude) mice. MetBo2 cells were injected into the mammary fat pad of female mice and, after 8 days, treated them daily with vehicle (3 mmol/L citrate buffer) or 40 mg/kg eCF506 by oral gavage. Animals' weight remained stable and comparable between the treatment groups during the length of the study (Supplementary Figs. S15A and S15B, Supplementary Materials and Methods). Administration of eCF506 was stopped after 28 days and tumor growth monitored in the absence of treatment until tumor relapse. As shown in Fig. 4D and E, potent antitumor activity was observed in all the mice treated with eCF506 from both immunocompetent (Fig. 4D) and immunocompromised (Fig. 4E) strains during the treatment phase, confirming the in vivo efficacy of eCF506 and the sensitivity of MetBo2 allografts to Src inhibition. Notably, major differences were observed during the posttreatment phase. Residual tumors regrew rapidly in CD1 nude mice after cessation of eCF506 treatment, undergoing >5-fold volume increment from days 28 to 39 (Fig. 4F). In contrast, tumor relapse was delayed by more than 11 days in immunocompetent mice following treatment cessation (Fig. 4D and F) and one mouse remained tumor-free until the end of the study (53 days), suggesting involvement of the immune system in the control of tumor growth during the posttreatment phase.

We show that eCF506 mediates very potent in vivo antitumor activity in murine triple-negative breast cancer models, against primary tumors and bone metastases, regardless of the mouse strain used. Nonetheless, we also show in the primary breast cancer model that significant differences emerge in the posttreatment period: mice with an intact immune system are capable to control tumor growth beyond the treatment phase (> 10 days), whereas tumor relapse occurs in immunosuppressed mice as soon as the animals are off treatment. Emulating the results of eCF506 in immunosuppressed mice, we show that dasatinib elicits a strong anticancer effect in immunocompetent mice during the treatment phase, but tumor growth accelerates immediately after halting the treatment. The posttreatment antitumor effect observed in the eCF506-treated arm (25% of mice remained tumor-free after the initial treatment phase) along with the higher response of most mice to the second treatment phase, resulted in significantly superior survival of the animals treated with eCF506 relative to dasatinib. Post-mortem analysis of the animals revealed smaller spleens in the dasatinib-treated group than in those treated with vehicle or eCF506. Notably, this effect was also observed in the short-term study (five daily doses). Taking into consideration that normal splenic histology was observed in all the treatment groups, dasatinib's effects are consistent with spleen hypoactivity (50). This is relevant because a reduction of tumor-fighting effector cells can affect the capacity of the adaptive immune system to respond to the cancer threat (58, 59). This is also supported by the histologic analysis of the tumors, which revealed a higher presence of peritumoral lymphocytes and CD3+ cells in the eCF506-treated mice compared with the dasatinib group and the vehicle control. However, the results from the short-term study suggests that macrophage reprogramming may be involved in the initiation of the differentiating phenotypic effects observed by eCF506 treatment. The overall pro-immune effects of eCF506 may be associated with its high selectivity over kinases that are targeted by dasatinib, such as ABL or KIT (see Supplementary Table S3). Although dasatinib has shown good tolerability in cancer patients (34), its broad immunosuppressive effects [which have been found to benefit CML patients by reducing regulatory T cells (60), but it also results in the suppression of cytotoxic T-cell function (50, 61) and myelosuppression (62)] is a potential disadvantage to treat solid malignancies in immunocompetent animals. Finally, we show that some of the animals of the dasatinib-treated group presented significant enlargement of the heart and damaged cardiac muscle tissue. This adverse effect, which is attributed to Abl inhibition (33, 34, 50), was not observed in the eCF506-treated group, which agrees with the superior safety profile of this selective inhibitor.

Influenza virus is a major health concern that has huge impacts on the human society, and vaccination remains as one of the most effective ways to mitigate this disease. Comparing the two types of commercially available Influenza vaccine, the live attenuated virus vaccine is more cross-reactive and easier to administer than the traditional inactivated vaccines. One promising live attenuated Influenza vaccine that has completed Phase I clinical trial is deltaFLU, a deletion mutant lacking the viral Nonstructural Protein 1 (NS1) gene. As a consequence of this gene deletion, this mutant virus can only propagate effectively in cells with a deficient interferon-mediated antiviral response. To demonstrate the manufacturability of this vaccine candidate, a batch bioreactor production process using adherent Vero cells on microcarriers in commercially available animal-component free, serum-free media is described.

Five commercially available animal-component free, serum-free media (SFM) were evaluated for growth of Vero cells in agitated Cytodex 1 spinner flask microcarrier cultures. EX-CELL Vero SFM achieved the highest cell concentration of 2.6 10^6 cells/ml, whereas other SFM achieved about 1.2 10^6 cells/ml. Time points for infection between the late exponential and stationary phases of cell growth had no significant effect in the final virus titres. A virus yield of 7.6 Log10 TCID50/ml was achieved using trypsin concentration of 10 μg/ml and MOI of 0.001. The Influenza vaccine production process was scaled up to a 3 liter controlled stirred tank bioreactor to achieve a cell density of 2.7 10^6 cells/ml and virus titre of 8.3 Log10 TCID50/ml. Finally, the bioreactor system was tested for the production of the corresponding wild type H1N1 Influenza virus, which is conventionally used in the production of inactivated vaccine. High virus titres of up to 10 Log10 TCID50/ml were achieved.

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We describe for the first time the production of Influenza viruses using Vero cells in commercially available animal-component free, serum-free medium. This work can be used as a basis for efficient production of attenuated as well as wild type Influenza virus for research and vaccine production.

Regardless of vaccine type (inactivated or live attenuated), virus vaccine production requires the initial step of propagating the Influenza viruses carrying the haemaglutinin and neuraminidase antigens of the strains that the vaccine is providing prophylaxis for. These viruses are traditionally propagated in embryonated hen eggs. Two important limitations of this process are the inflexible supply of high quality specific pathogen free (SPF) eggs and possible low titres of emerging viruses, such as the highly pathogenic Influenza A (H5N1) strain. To provide an alternative to egg-based vaccine production, mammalian cell culture based production has been developed in recent years [21]. This provides a flexible and scalable platform that can make use of existing biopharmaceutical infrastructure for Influenza vaccine production.

The growth kinetics of Vero cells in the 5 commercially available animal-component free, serum-free media (SFM) were evaluated in 250 ml spinner flasks. The media evaluated were OptiPro SFM (Invitrogen), VP-SFM (Invitrogen), EX-CELL Vero SFM (SAFC Bioscience), Provero-1 (Lonza) and HyQ SFM4MegaVir (HyClone). The results are presented in Figure 1A. Poor attachment of cells to microcarriers and poor cell growth was observed in HyQ SFM4MegaVir, which consequentially yielded a low cell concentration of 4.5 105 cells/ml. OptiPro SFM, VP-SFM and Provero-1 SFM displayed similar cell growth profiles, yielding cell concentrations of 1.2 106 cells/ml with viability above 90% on days 4 or 5. Growth of Vero cells in EX-CELL Vero SFM was the highest achieving 2.6 106 cells/ml (97% viability) on day 7.


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