NADPH phagocyte oxidase knockout mice control Trypanosoma cruzi proliferation, but develop circulatory collapse and succumb to infection

^•NO is considered to be a key macrophage-derived cytotoxic effector during Trypanosoma cruzi infection. On the other hand, the microbicidal properties of reactive oxygen species (ROS) are well recognized, but little importance has been attributed to them during in vivo infection with T. cruzi. In order to investigate the role of ROS in T. cruzi infection, mice deficient in NADPH phagocyte oxidase (gp91^{phox −/−} or phox KO) were infected with Y strain of T. cruzi and the course of infection was followed. phox KO mice had similar parasitemia, similar tissue parasitism and similar levels of IFN-γ and TNF in serum and spleen cell culture supernatants, when compared to wild-type controls. However, all phox KO mice succumbed to infection between day 15 and 21 after inoculation with the parasite, while 60% of wild-type mice were alive 50 days after infection. Further investigation demonstrated increased serum levels of nitrite and nitrate (NOx) at day 15 of infection in phox KO animals, associated with a drop in blood pressure. Treatment with a NOS2 inhibitor corrected the blood pressure, implicating NOS2 in this phenomenon. We postulate that superoxide reacts with ^•NO in vivo, preventing blood pressure drops in wild type mice. Hence, whilst superoxide from phagocytes did not play a critical role in parasite control in the phox KO animals, its production would have an important protective effect against blood pressure decline during infection with T. cruzi.     

Positive Selection Results in Frequent Reversible Amino Acid Replacements in the G Protein Gene of Human Respiratory Syncytial Virus

Human respiratory syncytial virus (HRSV) is the major cause of lower respiratory tract infections in children under 5 years of age and the elderly, causing annual disease outbreaks during the fall and winter. Multiple lineages of the HRSVA and HRSVB serotypes co-circulate within a single outbreak and display a strongly temporal pattern of genetic variation, with a replacement of dominant genotypes occurring during consecutive years. In the present study we utilized phylogenetic methods to detect and map sites subject to adaptive evolution in the G protein of HRSVA and HRSVB. A total of 29 and 23 amino acid sites were found to be putatively positively selected in HRSVA and HRSVB, respectively. Several of these sites defined genotypes and lineages within genotypes in both groups, and correlated well with epitopes previously described in group A. Remarkably, 18 of these positively selected tended to revert in time to a previous codon state, producing a “flip-flop” phylogenetic pattern. Such frequent evolutionary reversals in HRSV are indicative of a combination of frequent positive selection, reflecting the changing immune status of the human population, and a limited repertoire of functionally viable amino acids at specific amino acid sites.     

Investigations into,and development of,a lyophilized and formulated recombinant human factor IX produced from CHO cells

Objectives

To develop a recombinant human factor IX (rFIX) formulation equivalent to commercially available products in terms of cake appearance, residual moisture, proportion of soluble aggregates and activity maintenance for 3 months at 4–8 °C.

Results

NaCl and low bulking agent/cryoprotectant mass ratio had a negative impact on cake quality upon lyophilisation for a wide range of formulations tested. Particular devised formulations maintained rFIX activity after lyophilization with a similar performance when compared with the rFIX formulated using the excipients reported for a commercially available FIX formulation (Benefix). rFIX remained active after 3 months when stored at 4 °C, though this was not the case with samples stored at 40 °C. Interestingly, particular formulations had an increase in residual moisture after 3 months storage, but not above a 3% threshold. All four formulations tested were equivalent to the Benefix formulation in terms of particle size distribution and cake appearance.

Conclusions

Three specific formulations, consisting of surfactant polysorbate-80, sucrose or trehalose as cryoprotectant, mannitol or glycine as bulking agent, l-histidine as buffering agent, and NaCl added in the reconstitution liquid at 0.234% (w/v) were suitable for use with a CHO cell-derived recombinant FIX.

     

Development of a 3‐step straight‐through purification strategy combining membrane adsorbers and resins

The pressures to efficiently produce complex biopharmaceuticals at reduced costs are driving the development of novel techniques, such as in downstream processing with straight‐through processing (STP). This method involves directly and sequentially purifying a particular target with minimal holding steps. This work developed and compared six different 3‐step STP strategies, combining membrane adsorbers, monoliths, and resins, to purify a large, complex, and labile glycoprotein from Chinese hamster ovary cell culture supernatant. The best performing pathway was cation exchange chromatography to hydrophobic interaction chromatography to affinity chromatography with an overall product recovery of up to 88% across the process and significant clearance of DNA and protein impurities. This work establishes a platform and considerations for the development of STP of biopharmaceutical products and highlights its suitability for integration with single‐use technologies and continuous production methods. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:931–940, 2017     

Rethinking relevance: Repetition priming reveals the psychological reality of adaptive specializations for reasoning

Theories advanced to explain conditional reasoning range from those that invoke inference systems that evolved for specific domains (such as social exchange, precautions, or deontic regulations) to relevance theory, a relatively domain-general account that invokes conversational pragmatics. The present research utilized a novel extension of repetition priming, in conjunction with the Wason selection task (a widely known and used task to test people's conditional reasoning), to evaluate alternative theories of human reasoning. Across five experiments, testing over 600 participants, consistent priming across selection tasks was demonstrated. The pattern of priming effects supports models of human reasoning based on specific evolved reasoning abilities, and was inconsistent with general conditional reasoning models such as relevance theory. These results also converge with neurological and clinical evidence of divided psychological processes for reasoning about relatively specific domains, based on functionally distinct inference systems.     

Physically strong men are more militant: A test across four countries

There is substantial evidence from archaeology, anthropology, primatology, and psychology indicating that humans have a long evolutionary history of war. Natural selection, therefore, should have designed mental adaptations for making decisions about war. These adaptations evolved in past environments, and so they may respond to variables that were ancestrally relevant but not relevant in modern war. For example, ancestrally in small-scale combat, a skilled fighter would be more likely to survive a war and bring his side to victory. This ancestral regularity would have left its mark on modern men's intergroup psychology: more formidable men should still be more supportive of war. We test this hypothesis in four countries: Argentina, Denmark, Israel, and Romania. In three, physically strong men (but not strong women) were significantly more supportive of military action. These findings support the hypothesis that modern warfare is influenced by a psychology designed for ancestral war.     

Cytotoxicity and cellular accumulation of palladium(II) complexes of tetracyclines

We studied the cytotoxic effect and the uptake of Pd(II) complexes of doxycycline (Dox), [Pd(Dox)Cl2], and tetracycline (Tc), [Pd(Tc)Cl2], in chronic myelogenous leukemia cells. The effect of the compounds on macrophage viability was also investigated. Compound 1 is more effective than compound 2 in inhibiting the growth of K562 cells with the IC(50) values of 14.44 and 34.54 microM, respectively. There is a good correlation between cell-growth inhibition and intracellular metal concentrations, determined by inductively coupled plasma atomic emission spectroscopy (ICP-AES). Incubation of the cells with equitoxic concentrations of both compounds yields approximately the same intracellular Pd concentration. At the IC(50) doses, intracellular concentration is ca. 33 x 10(-16) mol/cell for both compounds 1 and 2. This suggests that more [Pd(Tc)Cl2] is needed to produce a cytotoxic effect, because it enters cells more slowly. Both compounds up to 16 microM did not affect the viability of mouse peritoneal macrophages after a 48-h incubation. After 72 h of incubation, the IC(50) values are 22 for [Pd(Dox)Cl2] and 40 microM for [Pd(Tc)Cl(2)]. Therefore, the cytotoxic effect in cancer cells exhibited by both compounds is higher than their effect in macrophages.     

Interaction between organophosphate insecticides and the parasitic mite Acarophenax lacunatus (Prostigmata: Acarophenacidae) on Rhyzopertha dominica (Coleoptera: Bostrichidae)

The interaction between the organophosphate insecticides fenitrothion and pirimiphos-methyl with Acarophenax lacunatus (Cross & Krantz) (Prostigmata: Acarophenacidae), an egg parasite of the stored grain pest Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae), was assessed in a range of doses for each compound. The number of physogastric females of A. lacunatus and egg parasitism (%) decreased with increasing insecticide doses and mite density for both organophosphates. Lower insecticide doses for fenitrothion and pirimiphos-methyl and lower mite densities led to higher instantaneous rates of increase of the mite population. Overall lower instantaneous rates of increase of A. lacunatus were obtained with the insecticide pirimiphos-methyl. The sustained presence of the mite species in all insecticide doses suggest that this biological control agent may be used together with insecticide applications for controlling R. dominica, but high insecticide doses, mainly of pirimiphos-methyl, compromise mite population growth. The high rates of egg parasitism obtained with the biological control agent may aid the insecticide effect on the adult pest population preventing its outbreaks in wheat grains, constituting a potential tactic for pest management of stored wheat.