Some Properties of an Extracellular Proteolytic Enzyme of Verticillium fungicola, a Pathogen of the Cultivated Mushroom Agaricus bisporus

In a casein nutrient solution, Verticillium fungiocla, the causal agent of the dry bubble disease of the cultivated mushroom Agaricus bisporus, produces a proteolytic enzyme. The effects of the pH and of inhibitors on the protease activity and the heat stability of the enzyme are described. The protease is of interest in connection with the attacking mechanism of Verticillium fungicola.     

Cellular immunotherapy with multiple infusions of in vitro-expanded haploidentical natural killer cells after autologous transplantation for patients with plasma cell myeloma

Background aimsTo investigate the feasibility and safety of haploidentical natural killer (NK) cell infusions as consolidation immunotherapy after autologous stem cell transplant (ASCT) in patients with plasma cell myeloma.MethodsTen patients (median age, 59 years) received induction treatment followed by high-dose melphalan (200 mg/m²) at day –1, ASCT at day 0 and increasing NK cell doses (1.5 × 10⁶, 1.5 × 10⁷ and multiple doses of 1.0 × 10⁸ cells/kg body weight) from day +1 to day +30 after ASCT. NK cells were harvested and purified from peripheral blood of haploidentical donors and expanded for 19 days with interleukin (IL)-2 and IL-15 under Good Manufacturing Practice conditions.ResultsNK cell numbers increased 56.0-fold (37.4- to 75.5-fold). Patients received a median of 3.8 × 10⁸ (0.9–5.7 × 10⁸) NK cells/kg body weight in six (three to eight) infusions. Multiparametric mass cytometry analysis demonstrated an altered surface receptor repertoire of expanded NK cells with increased degranulation and cytokine production activities but diminished expression of perforin. Donor NK cells were detectable in the peripheral blood, peaking 1 h after each dose (up to 90% donor NK cells). The treatment was safe and well tolerated, without evidence of graft-versus-host disease. Comparison with a control patient population receiving ASCT without NK cell infusions showed no significant difference in relapse, progression-free survival and overall survival.ConclusionsThis study demonstrates reliable manufacturing of high numbers of activated NK cells for multiple-dose infusions and safe administration of these cellular products. The trial was registered at ClinicalTrials.gov (identifier no. NCT01040026).     

The root–soil system of Norway spruce subjected to turning moment: resistance as a function of rotation

The reactions of trees to wind, rockfall, and snow and debris flow depend largely on how strong and deformable their anchorage in the soil is. Here, the resistive turning moment M of the root–soil system as a function of the rotation ϕ at the stem base plays the major role. M(ϕ) describes the behavior of the root–soil system when subject to rotational moment, with the maximum M(ϕ) indicating the anchorage strength M _a of the tree. We assessed M(ϕ) of 66 Norway spruce (Picea abies L. Karst) by pulling them over with a winch. These 45- to 170-year-old trees grew at sites of low and high elevation, with a diameter at breast height DBH = 14–69 cm and a height H = 9–42 m. M(ϕ) displayed a strong nonlinear behavior. M _a was reached at a lower ϕ for large trees than for small trees. Thus overhanging tree weight contributed less to M _a for the large trees. Overturning also occurred at a lower ϕ for the large trees. These observations show that the rotational ductility of the root–soil system is higher for small trees. M _a could be described by four monovariate linear regression equations of tree weight, stem weight, stem volume and DBH ² ·H (0.80 < R ² < 0.95), and ϕ at M _a, ϕ _a, by a power law of DBH²·H (R ² = 0.85). We found significantly higher M _a for the low-elevation spruces than for the high-elevation spruces, which were more shallowly anchored, but no significant difference in ϕ _a. The 66 curves of M(ϕ), normalized (_n) by M _a in M-direction and by ϕ _a in ϕ-direction, yielded one characteristic average curve: . Using and the predictions of M _a and ϕ _a, it is shown that M(ϕ) and the curves associated with M(ϕ) can be predicted with a relative standard error ≤25%. The parameterization of M(ϕ) by tree size and weight is novel and provides useful information for predicting with finite-element computer models how trees will react to natural hazards.     

Evolution of age at primiparity in pinnipeds in the absence of the quality–quantity trade‐off in reproduction

Age at primiparity (AP) is a key life history trait which is crucial to the evolution of life history strategies. This trait is particularly interesting in pinnipeds (walrus, eared seals, and true seals), which are monotocous animals. Thus, the commonly observed trade‐off between offspring quality and quantity does not apply to this taxon. Therefore, comparative studies on the evolution of AP might shed light on other important evolutionary correlates when litter size is fixed. Using phylogenetic generalized least squares analyses, we found a strong negative and robust correlation between relative birth mass (mean pup birth mass as a proportion of mean adult female mass) and AP. Rather than trading‐off an early start of reproduction with light relative offspring mass, this result suggests that pinnipeds exhibit either faster (i.e., higher relative offspring mass leading to shorter lactation length, and thus shorter interbirth interval) or slower life histories and that an early AP and a heavy relative offspring mass co‐evolved into a comparatively fast life history strategy. On the other hand, AP was positively related to lactation length: A later start of reproduction was associated with a longer lactation length. Consequently, variation in AP in pinnipeds seems to be affected by an interplay between costs and benefits of early reproduction mediated by relative investment into the single offspring via relative birth mass and lactation length.     

Natural frequencies and damping ratios of Norway spruce (<Emphasis Type="Italic">Picea abies</Emphasis> (L.) Karst) growing on subalpine forested slopes

We investigated the natural frequencies and damping ratios of 24 Norway spruce trees (Picea abies (L.) Karst) growing on subalpine forested slopes using swaying experiments. Trees were winched to the release position using a manually operated cable winch. Before release, the stem base rotation was approximately 2 degrees. All trees were tested in both fall-line and cross-slope direction if possible to reveal the difference between the two perpendicular directions. A new method based upon the Hilbert transformation for evaluating the damping ratio for trees was used. It enabled us to investigate the damping ratio for all excited modes of vibration in the tree structure during the swaying experiments and to study if velocity proportional damping (viscous damping) is a reasonable assumption when predicting the tree response to different external actions using mechanical models. No difference in first natural frequency and damping ratio was found between the fall-line and cross-slope direction for all tested trees. The first natural frequencies correlated best with the diameter at breast height and squared total tree height DBH/H ². For the damping ratio, no significant correlation with different tree characteristics could be found. A non-velocity proportional damping ratio was observed for two trees. This indicates that using a viscous damping for all frequencies is an appropriate simplification when modelling the wind-tree interaction.     

Formalin inactivation of vesicular stomatitis virus impairs T-cell- but not T-help-independent B-cell responses.

The effects of formalin on the infectivity and immunogenicity of vesicular stomatitis virus (VSV) serotype Indiana were investigated. We found that formalin inactivation of VSV prevents infection of Vero cells in a concentration- and time-dependent manner, as shown by fluorometric cell analysis and inhibition of plaque formation. Inactivated VSV failed to induce significant cytotoxic T-lymphocyte responses in vivo or after restimulation in vitro. In contrast, the early immunoglobulin M (IgM) response, which is T help independent in the VSV system, was unaltered, suggesting normal antigenicity for and induction of B cells. However, no switch to IgG occurred, demonstrating failure of induction of T help. If cross-reactive T help was provided by previous infection with a second serotype of VSV (New Jersey), the IgG response was almost completely restored, confirming that the absence of IgG was due to lack of T help. A formalin-treated preparation of glycoprotein of VSV led to a delayed but otherwise normal IgG response, whereas treatment of VSV with UV light or beta-propiolactone reduced IgG titers to the same extent as did formalin. These results suggest that loss of infectivity and the ensuing lack of amplification of viral antigens of formaldehyde-inactivated VSV is the major factor impairing induction of specific T-helper cell responses.