Depletion of extrinsic pathway inhibitor (EPI) sensitizes rabbits to disseminated intravascular coagulation induced with tissue factor: evidence supporting a physiologic role for EPI as a natural anticoagulant.

Although in vitro experiments have established that extrinsic pathway inhibitor (EPI) is the only known plasma inhibitor of factor VIIa-tissue factor (TF) catalytic activity of potential physiologic significance, evidence of its function in vivo has been lacking. TF-induced intravascular coagulation may occur in patients despite normal plasma levels of EPI and, in our earlier studies, normal plasma EPI levels did not protect rabbits from intravascular coagulation induced by an infusion of purified TF (1 microgram/kg). Studies have now been carried out in which plasma EPI levels were reduced in rabbits to below 20% of the initial level by injection of anti-rabbit EPI IgG. Infusion into such animals of purified rabbit TF apoprotein (0.25 microgram/kg) reconstituted into phospholipid vesicles induced substantial disseminated intravascular coagulation. Infusion of control saline or phospholipid vesicles not containing TF was without significant effect as was infusion of TF (0.25 microgram/kg) into animals injected with nonimmune goat IgG. These data establish that EPI can dampen TF-induced intravascular coagulation in rabbits. They support the hypothesis that EPI plays a significant role in regulating coagulation resulting from the exposure of blood to trace concentrations of TF during the illnesses and minor injuries of normal existence.     

The origin recognition complex is dispensable for endoreplication in Drosophila

The origin recognition complex (ORC) is an essential component of the prereplication complex (pre-RC) in mitotic cell cycles. The role of ORC as a foundation to assemble the pre-RC is conserved from yeast to human. Furthermore, in metazoans ORC plays a key role in determining the timing of replication initiation and origin usage. In this report we have produced and analyzed a Drosophila orc1 allele to investigate the roles of ORC1 in three different modes of DNA replication during development. As expected, ORC1 is essential for mitotic replication and proliferation in brains and imaginal discs, as well as for gene amplification in ovarian follicle cells. Surprisingly, however, ORC1 is not required for endoreplication. Decreased cell number in orc1 mutant salivary glands is consistent with the idea that undetectable levels of maternal ORC1 during embryogenesis fail to support further proliferation. Nevertheless, these cells begin endoreplicating normally and reach a final ploidy of >1000C in the absence of zygotic synthesis of ORC1. The dispensability of ORC is further supported by an examination of other ORC members, whereas Double-parked protein/Cdt1 and minichromosome maintenance proteins are apparently essential for endoreplication, implying that some aspects of initiation are shared among the three modes of DNA replication. This study provides insight into the physiologic roles of ORC during metazoan development and proposes that DNA replication initiation is governed differently in mitotic and endocycles.     

Development of free-living stages of Strongyloides ratti under different temperature conditions

It is well known that the Strongyloides species have two different developmental courses—direct and indirect development—and selection of these courses is affected by various environmental factors. This study examined the effect of temperature on the development of first-stage larvae (L1s) of Strongyloides ratti, to clarify how larvae adapt and survive at unsuitable temperatures. It was revealed that L1s cultured at 4 or 10 °C for 120 h could not develop because of growth arrest or delay. However, L1s could develop after transfer to culture at 25 °C for 48 h. Although larvae cultured at 25 °C take indirect development, larvae subjected to low-temperature stimulation (at 4 or 10 °C) take direct development into infective third-stage larvae (L3s), and only 1 min of low-temperature stimulation was sufficient to induce direct development. Morphological study of low-temperature-stimulated L3s revealed that those stimulated at 4 °C (L3–4) showed less development, but those stimulated at 10 °C (L3–10) developed as well as the control (no low-temperature stimulation). Furthermore, we revealed that L3–10 showed similar infectivity to the control when they were injected subcutaneously into rats as the final host, which indicated that L3–10 grew normally. We conclude that S. ratti has a survival strategy of growth arrest or delay if excreted in cold conditions. Moreover, even if they start development after transfer to suitable conditions, they memorize low-temperature stimulation, which leads them to direct development thereafter so that they can immediately infect the final host.     

Bottom-up analysis of emergent properties of N-acetylcysteine as an adjuvant therapy for COVID-19

N-acetylcysteine (NAC) is an abundantly available antioxidant with a wide range of antidotal properties currently best studied for its use in treating acetaminophen overdose. It has a robustly established safety profile with easily tolerated side effects and presents the Food and Drug Administration's approval for use in treating acetaminophen overdose patients. It has been proven efficacious in off-label uses, such as in respiratory diseases, heart disease, cancer, human immunodeficiency virus infection, and seasonal influenza. Clinical trials have recently shown that NAC's capacity to replenish glutathione stores may significantly improve coronavirus disease 2019 (COVID-19) outcomes, especially in high risk individuals. Interestingly, individuals with glucose 6-phosphate dehydrogenase deficiency have been shown to experience even greater benefit. The same study has concluded that NAC's ability to mitigate the impact of the cytokine storm and prevent elevation of liver enzymes, C-reactive protein, and ferritin is associated with higher success rates weaning from the ventilator and return to normal function in COVID-19 patients. Considering the background knowledge of biochemistry, current uses of NAC in clinical practice, and newly acquired evidence on its potential efficacy against COVID-19, it is worthwhile to investigate further whether this agent can be used as a treatment or adjuvant for COVID-19.     

Comparison of Torque,Screw-in Force,and Shaping Ability of Glide Path Instruments in Continuous Rotation and Optimum Glide Path Motion

IntroductionThis study aimed to analyze torque/force generation and transportation in double-curved canals instrumented with 3 types of glide path files using optimum glide path (OGP) motion in comparison with continuous rotation.MethodsSixty simulated double-curved canals were prepared with #10/0.05 or #15/0.03 HyFlex EDM Glidepath files (Coltene/Whaledent, Altstätten, Switzerland) or a #13/0.04 prototype MANI Glidepath file (Tochigi, Japan) using OGP motion or continuous rotation (n = 10 each). Canals were sequentially prepared to 20 mm and 22 mm (full working length) using automated root canal instrumentation and a torque/force analyzing device. Transportation was calculated at 1–9 mm from the apex. Data were compared using 2-way analysis of variance followed by a post hoc simple main effect test with Bonferroni correction and a Kruskal-Wallis test (α = 5%).ResultsAll #10/0.05 instruments fractured. In the 22-mm preparation, the OGP motion resulted in lower clockwise torque and screw-in force than did continuous rotation (P < .05). In the 20-mm preparation, #15/0.03 instruments recorded a lower screw-in force for OGP motion than for continuous rotation (P < .05). Comparing the 2 preparation phases, OGP motion generated no significant differences; however, continuous rotation developed higher clockwise torque and screw-in force in the 22-mm preparation than in the 20-mm preparation (P < .05). There was no significant difference among the tested groups for transportation values.ConclusionsCompared with continuous rotation, OGP motion generated less screw-in force, lower clockwise torque, and similar transportation. The #15/0.03 HyFlex EDM instrument and the #13/0.04 prototype MANI instrument performed similarly well.     

Severe cellulitis caused by Achromobacter xylosoxidans after allogeneic hematopoietic stem cell transplantation

Achromobacter xylosoxidans (A. xylosoxidans) is an aerobic gram-negative bacillus and often isolated from aquatic environments. It is supposed to cause infections in patients with malignancy or immunodeficiency. It causes various healthcare-associated infections, but cellulitis is rare. Herein, we report the first case of sever cellulitis by A. xylosoxidans after allogeneic hematopoietic stem cell transplantation (HSCT). A 49-year-old man underwent allogeneic HSCT from 8/8 HLA-matched unrelated donor with myeloablative conditioning for relapsed acute myeloid leukemia. He developed skin chronic graft versus host disease 11 months after HSCT. During the prolonged treatment with prednisolone and cyclosporine, he developed cellulitis on his left leg and admitted to our hospital. Blood and exudate culture revealed A. xylosoxidans. Although empirical therapy with cefepime was ineffective, his symptoms were dramatically improved after administration of meropenem. To our knowledge, this is the first case of A. xylosoxidans cellulitis after allogeneic HSCT. A. xylosoxidans should be considered as a possible cause of cellulitis in post-allogeneic HSCT patients on prolonged immunosuppressive therapy.