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Blood-feeding arthropods secrete potent salivary molecules, which include platelet aggregation inhibitors, vasodilators, and anticoagulants. Among these molecules, Alboserpin, the major salivary anticoagulant from the mosquito vector Aedes albopictus , is a specific inhibitor of the human coagulation factor Xa (FXa). In this study, we investigated the anti-inflammatory properties of Alboserpin, in vitro and in vivo. In vitro, Alboserpin inhibited FXa-induced protease-activated receptor (PAR)-1, PAR-2, PAR-3, VCAM, ICAM, and NF-κB gene expression in primary dermal microvascular endothelial cells. Alboserpin also prevented FXa-stimulated ERK1/2 gene expression and subsequent inflammatory cytokine release (MCP-1, TNF-α, IL-6, IL-8, IL-1β, IL-18). In vivo, Alboserpin reduced paw edema induced by FXa and subsequent release of inflammatory cytokines (CCL2, MCP-1, IL-1α, IL-6, IL-1β). Alboserpin also reduced FXa-induced endothelial permeability in vitro and in vivo. These findings show that Alboserpin is a potent anti-inflammatory molecule, in vivo and in vitro, and may play a significant role in blood feeding., (Copyright © 2022 The Authors.)

Phosphatidyl inositol (4,5)-bisphosphate (PI(4,5)P 2 ) plays several key roles in human biology and the lipid kinase that produces PI(4,5)P 2 , PIP5K, has been hypothesized to provide a potential therapeutic target of interest in the treatment of cancers. To better understand and explore the role of PIP5K in human cancers there remains an urgent need for potent and specific PIP5K inhibitor molecules. Following a high throughput screen of the AstraZeneca collection, a novel, moderately potent and selective inhibitor of PIP5K, 1, was discovered. Detailed exploration of the SAR for this novel scaffold resulted in the considerable optimization of both potency for PIP5K, and selectivity over the closely related kinase PI3Kα, as well as identifying several opportunities for the continued optimization of drug-like properties. As a result, several high quality in vitro tool compounds were identified (8, 20 and 25) that demonstrate the desired biochemical and cellular profiles required to aid better understanding of this complex area of biology., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Female mosquitoes require blood meals for egg development. The saliva of blood feeding arthropods contains biochemically active molecules, whose anti-hemostatic and anti-inflammatory properties facilitate blood feeding on vertebrate hosts. While transcriptomics has presented new opportunities to investigate the diversity of salivary proteins from hematophagous arthropods, many of these proteins remain functionally undescribed. Previous transcriptomic analysis of female salivary glands from Culex quinquefasciatus, an important vector of parasitic and viral infections, uncovered a 12-member family of putatively secreted proteins of unknown function, named the Cysteine and Tryptophan-Rich (CWRC) proteins. Here, we present advances in the characterization of two C. quinquefasciatus CWRC family members, CqDVP-2 and CqDVP-4, including their enrichment in female salivary glands, their specific localization within salivary gland tissues, evidence that these proteins are secreted into the saliva, and their native crystal structures, at 2.3 ​Å and 1.87 ​Å, respectively. The β-trefoil fold common to CqDVP-2 and CqDVP-4 is similar to carbohydrate-binding proteins, including the B subunit of the AB toxin, ricin, from the castor bean Ricinus communis . Further, we used a glycan array approach, which identifies carbohydrate ligands associated with inflammatory processes and signal transduction. Glycan array 300 testing identified 100 carbohydrate moieties with positive binding to CqDVP-2, and 77 glycans with positive binding to CqDVP-4. The glycan with the highest relative fluorescence intensities, which exhibited binding to both CqDVP-2 and CqDVP-4, was used for molecular docking experiments. We hypothesize that these proteins bind to carbohydrates on the surface of cells important to host immunology. Given that saliva is deposited into the skin during a mosquito bite, and acts as the vehicle for arbovirus inoculation, understanding the role of these proteins in pathogen transmission is of critical importance. This work presents the first solved crystal structures of C. quinquefasciatus salivary proteins with unknown function. These two molecules are the second and third structures reported from salivary proteins from C. quinquefasciatus , an important, yet understudied disease vector., Competing Interests: There is no conflict of interest.

Aedes aegypti saliva facilitates blood meal acquisition through pharmacologically active compounds that prevent host hemostasis. Among these salivary proteins are the D7s, which are highly abundant and have been shown to act as scavengers of biogenic amines and eicosanoids. In this work, we performed comparative structural modeling, characterized the binding capabilities, and assessed the physiological functions of the Ae. aegypti salivary protein AeD7L2 compared to the well-characterized AeD7L1. AeD7L1 and AeD7L2 show different binding affinities to several biogenic amines and biolipids involved in host hemostasis. Interestingly, AeD7L2 tightly binds U-46619, the stable analog of thromboxane A 2 (K D  = 69.4 nm), which is an important platelet aggregation mediator, while AeD7L1 shows no binding. We tested the ability of these proteins to interfere with the three branches of hemostasis: vasoconstriction, platelet aggregation, and blood coagulation. Pressure myography experiments showed these two proteins reversed isolated resistance artery vasoconstriction induced by either norepinephrine or U-46619. These proteins also inhibited platelet aggregation induced by low doses of collagen or U-46619. However, D7 long proteins did not affect blood coagulation. The different ligand specificity and affinities of AeD7L1 and AeD7L2 matched our experimental observations from studying their effects on vasoconstriction and platelet aggregation, which confirm their role in preventing host hemostasis. This work highlights the complex yet highly specific biological activities of mosquito salivary proteins and serves as another example of the sophisticated biology underlying arthropod blood feeding., (Published 2020. This article is a U.S. Government work and is in the public domain in the USA. The FEBS Journal published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)

During blood-feeding, mosquito saliva is injected into the skin to facilitate blood meal acquisition. D7 proteins are among the most abundant components of the mosquito saliva. Here we report the ligand binding specificity and physiological relevance of two D7 long proteins from Culex quinquefasciatus mosquito, the vector of filaria parasites or West Nile viruses. CxD7L2 binds biogenic amines and eicosanoids. CxD7L1 exhibits high affinity for ADP and ATP, a binding capacity not reported in any D7. We solve the crystal structure of CxD7L1 in complex with ADP to 1.97 Å resolution. The binding pocket lies between the two protein domains, whereas all known D7s bind ligands either within the N- or the C-terminal domains. We demonstrate that these proteins inhibit hemostasis in ex vivo and in vivo experiments. Our results suggest that the ADP-binding function acquired by CxD7L1 evolved to enhance blood-feeding in mammals, where ADP plays a key role in platelet aggregation.

The mass mortality of oak trees has been prevalent in Japan since the late 1980s. The fungus Raffaelea quercivora is transmitted by an ambrosia beetle, Platypus quercivorus, which causes mortality. The beetle is able to bore galleries into the sapwood of most Fagaceae trees in Japan; however, the level of mortality caused by R. quercivora and P. quercivorus differs greatly among tree species. Previous studies by our research group have demonstrated that the virulence of R. quercivora differs among isolates when inoculated into Quercus serrata logs. However, interactions between the virulence of R. quercivora isolates and the susceptibility of other fagaceous species have yet to be elucidated. In this study, we inoculated the fresh logs of 11 fagaceous species with isolates of low and high virulence, and measured the tangential widths of discolored sapwoods 3 weeks after inoculation. Although the discoloration widths of Q. crispula sapwood were similar among all isolates, those of Q. serrata and Q. acutissima tended to increase with the more virulent isolates. Sapwood discoloration in Q. glauca, Q. acuta, Q. salicina, Lethocarpus edulis, and Castanopsis sieboldii was greatly increased by highly virulent isolates. Discoloration in Fagus japonica was not influenced by any of the isolates. The logs of Q. crispula and Q. serrata but not Q. glauca were significantly more discolored by a low-virulence isolate compared with standing trees. The various virulent isolates induced unique sapwood discoloration characteristics in each species, which may explain species-specific differences in mortality rates.

Centrosomes associate with spindle poles; thus, the presence of two centrosomes promotes bipolar spindle assembly in normal cells. Cancer cells often contain supernumerary centrosomes, and to avoid multipolar mitosis and cell death, these are clustered into two poles by the microtubule motor protein HSET. We report the discovery of an allosteric inhibitor of HSET, CW069, which we designed using a methodology on an interface of chemistry and biology. Using this approach, we explored millions of compounds in silico and utilized convergent syntheses. Only compound CW069 showed marked activity against HSET in vitro. The inhibitor induced multipolar mitoses only in cells containing supernumerary centrosomes. CW069 therefore constitutes a valuable tool for probing HSET function and, by reducing the growth of cells containing supernumerary centrosomes, paves the way for new cancer therapeutics., (Copyright © 2013 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Little information is available describing how to implement a disease registry from an electronic patient record system. The aim of this report is to describe the technology, methods, and utility of a diabetes registry populated by the Veterans Health Information Systems Architecture (VistA), which underlies the computerized patient record system of the Veterans Health Administration (VHA) in Veteran Affairs Integrated Service Network 10 (VISN 10)., Methods: VISN 10 data from VistA were mapped to a relational SQL-based data system using KB_SQL software. Operational definitions for diabetes, active clinical management, and responsible providers were used to create views of patient-level data in the diabetes registry. Query Analyzer was used to access the data views directly. Semicustomizable reports were created by linking the diabetes registry to a Web page using Microsoft asp.net2. A retrospective observational study design was used to analyze trends in the process of care and outcomes., Results: Since October 2001, 81,227 patients with diabetes have enrolled in VISN 10: approximately 42,000 are currently under active management by VISN 10 providers. By tracking primary care visits, we assigned 91% to a clinic group responsible for diabetes care. In the Cleveland Veterans Affairs Medical Center (VAMC), the frequency of mean annual hemoglobin A1c levels > or =9% has declined significantly over 5 years. Almost 4000 patients have been seen in diabetes intervention programs in the Cleveland VAMC over the past 4 years., Conclusions: A diabetes registry can be populated from the database underlying the VHA electronic patient record database system and linked to Web-based and ad hoc queries useful for quality improvement.

A dynamical decoupling method is presented which is based on embedding a deterministic decoupling scheme into a stochastic one. This way it is possible to combine the advantages of both methods and to increase the suppression of undesired perturbations of quantum systems significantly even for long interaction times. As a first application the stabilization of a quantum memory is discussed which is perturbed by one- and two-qubit interactions.

Elective orthopaedic surgery is regularly withheld from patients with haemophilia and high inhibitor titre despite the presence of severe arthropathy and urgent medical need. A knee joint arthroplasty was performed in a patient with severe haemophilia A and a high inhibitor titre using recombinant factor VIIa (rFVIIa) as the sole coagulation factor. There was no abnormal bleeding during surgery although an increased blood loss through surgical drains did occur during the first 6 h postoperatively. Rehabilitation was started on day 1 and continued for 3 months. Walking commenced on day 4. After 1 year of follow-up, the clinical outcome of surgery was considered excellent with no pain, knee mobility at 0-5-90 degrees, and an International Knee Society score of 95/100. No rFVIIa-associated side-effects or thrombotic complications were reported. In conclusion, knee joint arthroplasty is now an option for haemophilia patients with a high inhibitor titre. An international review of all available data on elective orthopaedic surgery in inhibitor patients is required so that the optimal treatment regime can be defined and the short- and long-term risk-benefit ratio of surgery compared to that of noninhibitor patients.

Background: Brachial plexus is usually approached by the supraclavicular or axillary route. A technique for selective blockade of the branches of the plexus at the humeral canal using electrolocation has recently been proposed. The aim of the present study was to assess the feasibility of this technique in the ambulatory patient and to determine the optimal sequence of nerve-blocking., Methods: The nerves originating from the brachial plexus were located in the humeral canal, at the junction of the proximal and the middle third of the arm, with a stimulator and blocked using either lidocaine or a mixture of lidocaine and bupivacaine, depending on the anticipated duration of surgery. The minimal stimulating intensity eliciting an adequate response, type of local anaesthetic and injected volume, and time of onset of surgical anaesthesia were collected., Results: The study included 503 consecutive ambulatory patients due to undergo surgery of the elbow, wrist or hand in one year. Suitable anaesthesia was obtained with the humeral blockade in 82.1% of cases. In the remaining 17.9%, an additional block at the elbow was required, mainly for ulnar and median nerves. The onset times of sensory blocks were the longest for the median nerve, similar for the radial and ulnar nerves, shorter for the musculocutaneous nerve and the shortest for the medial brachial and antebrachial cutaneous nerves. The difference was more significant with the lidocaine-bupivacaine mixture, than with lidocaine alone (P<0.001 vs P<0.05, respectively). The onset times of motor blocks were the longest for the median nerve (P<0.05) and the shortest for the musculocutaneous nerve (P<0.001). Neither nervous nor vascular complications occurred., Conclusion: This study shows that the nerve block at the humeral canal is an efficient and safe technique. Considering the onset times of nerve blocks, the following sequence for blockade can be recommended: median, ulnar, radial, musculocutaneous, medial (brachial and antebrachial) cutaneous nerves. The selective blockade of the main nerves of the upper limb at the humeral canal can be recommended for surgery of the forearm and the hand in the ambulatory patient.

Besides the microsurgical ventral decompression for treatment of cervical disc prolapses, combined with an intercorporal fusion using an autologous bone graft from the iliac crest and a plate osteosynthesis, the microsurgical, transoral approach to the craniocervical junction has proven to be an effective procedure for adequate indications. Even for surgical treatment of diseases and injuries of the craniocervical junction ventral, anterolateral, lateral and dorsal approaches are applicable alone or in combination. The special anatomic and functional conditions of this region, however, obviously require that the indicational criteria for the various approaches differ from those selected for the other cervical segments. The postoperative complication risk requires that particulary critical consideration be given to the question of isolated transoral interventions. The same holds true for the question as the necessity for additional ventral stabilisation in combined dorsoventral interventions. This report is about 20 patients who underwent transoral decompression, about the indications and the procedure typical problems.

Various therapeutic options have been proposed in the treatment of femoral head necrosis ranging from conservative management to total hip arthroplasty. Since microsurgical techniques are routinely used in orthopaedic surgery, the importance of revascularization has increased in recent years. Vascularized bone grafting as well as various osteotomies have been considered for the stages II and III according to Ficat and II, III and IV of the criteria of the ARCO system respectively, which also bases on MRI-findings. We investigated the results of 80 patients with avascular necrosis of the femoral head treated with a vascular pedicled iliac bone graft, perfused by the A. circumflexa ilium profunda, between 1988 and 1996. Mean follow-up was 5.6 years. The evaluation was based on the harris hip score, clinical and radiological examination as well as a subjective assessment using a VAS (visual analog scale). The clinical results according to the harris hip score were good or excellent in 86.1%. These results correlate with the subjective assessment of pain and of the hip joint function with an average of 7.9 points using the VAS (max. 10 points). Unchanged radiological appearance over the follow-up period was observed in 47.5% using the ARCO criterias. Reviewing the studies using vascularized grafts, about 50% of the patients with a stage II and III according to Ficat and II, III and IV of the ARCO respectively show an unchanged stage of the disease 5-6 years after the procedure. Therefore, transplantation of a vascular pedicled iliac bone graft possibly offers the chance to intervene causally in the course of the disease with only little alteration of the biomechanics of the hip joint.

Objective: To analyse cases of Horner's syndrome (HS) and its opposite, Pourfour du Petit's syndrome (PPS), occurring in anaesthesia and intensive therapy with consideration of the data of current literature., Data Sources: For this paper we have reviewed the French, English and German literature published in anaesthesia and intensive care journals using Medline search and the current textbooks., Study Selection: All observational studies on these syndromes, whether clinical cases or letters to the editor, form the basis for this article., Data Extraction: The articles were analysed mainly with regard to diagnosis, therapy and prognosis of syndromes due to iatrogenic causes., Data Synthesis: HS is caused by a paralysis of the ipsilateral sympathetic cervical chain and includes a ptosis of the upper eyelid, a slight elevation of the lower lid, a sinking of the eyeball, a constriction of the pupil, a narowing of the palpebral fissure, a nasal stuffiness associated with anhidrosis, and flushing of the affected side of the face. Regional anaesthesia (intra-oral anaesthesia, brachial plexus block, epidural anaesthesia whether by thoracic, lumbar or caudal approach, as well as interpleural analgesia) is the main anaesthetic cause for HS. HS due to the effect of a local anaesthetic is transient, it can precede a high spinal block and a cardiovascular collapse. HS from puncture of the internal jugular vein is most often permanent. When transient, HS regresses within 3 months after puncture. Other causes of HS include intraoperative posture, pleural drain, neck surgery, neck trauma. A mydriatic collyrium, such as phenylephrine, resolves ptosis for less than 1 hour and results in blurred vision from pupillary dilation. Major ptosis requires surgery. PPS is the reciprocal HS and is caused by a stimulation of the ipsilateral sympathetic cervical chain. PPS can precede HS. It carries a risk for conjunctivitis, keratitis and epiphora in case of major exophthalmia. PPS is often reported as an unilateral mydriasis. PPS has the same causes as HS. Myotic collyriums are relatively inefficient. Major lid retraction requires a tarsorraphy, pomades and nocturnal lid occlusion. A part of HS and most PPS occurring in anaesthesia and intensive care remain unrecognized or are recognized with delay, especially if they remain minor and transient or when they occur in unconscious patients, in horizontal posture.

We are reporting our experience with the efficacy of free tissue transfer in treatment of chronic lower-extremity osteomyelitis; 23 latissimus dorsi flaps and 11 rectus abdominis flaps were used. In our series, success was achieved after a mean follow-up of 36.1 months in 79% and after additional wound debridement in 94%. Indications, methods, results and complications are reported.

In rats a severe but compensated chronic renal insufficiency was induced by stepwise 9/10 nephrectomy. Despite this severe chronic renal insufficiency we observed no relevant pathological changes in the intestinal mucosa. In particular, we found no evidence of mucosal erosions, ulceration or pseudomembranous colitis, findings which are traditionally thought to be characteristic of the uremic state. This was also true of those animals dying prematurely from uremia. Thus serious doubts arise about the existence of "uremic enterocolitis", doubts which also proved justified after a critical review of the literature on human pathology.