Umbilical cord clamping in the early phases of the COVID-19 era - a systematic review and meta-analysis of reported practice and recommendations in guidelines.

OBJECTIVES: At the beginning of the COVID-19 pandemic, delayed umbilical cord clamping (CC) at birth may have been commonly discouraged despite a lack of convincing evidence of mother-to-neonate SARS-CoV-2 transmission. We aimed to systematically review guidelines, and reports of practice and to analyze associations between timing of CC and mother-to-neonate SARS-CoV-2 transmission during the early phases of the pandemic. METHODS: Major databases were searched from December 1, 2019, to July 20, 2021. INCLUSION: studies and guidelines describing CC practice in women with SARS-CoV-2 infection during pregnancy until 2 postnatal days, giving birth to live-born neonates. EXCLUSION: no extractable data. Two reviewers independently screened studies for eligibility and assessed study quality. Pooled prevalence rates were calculated. RESULTS: Forty-eight studies (1476 neonates) and 40 guidelines were included. Delayed CC was recommended in 70.0% of the guidelines. Nevertheless, delayed CC was reported less often than early CC: 262/1476 (17.8%) vs 511/1476 (34.6%). Neonatal SARS-CoV-2 positivity rates were similar following delayed (1.2%) and early CC (1.3%). Most SARS-CoV-2 transmissions (93.3%) occurred in utero. CONCLUSION: Delayed CC did not seem to increase mother-to-neonate SARS-CoV-2 transmission. Due to its benefits, it should be encouraged even in births where the mother has a SARS-CoV-2 infection. SYSTEMATIC REVIEW REGISTRATION: Prospero CRD42020199500.

[Spasticity-correcting surgery can have long-lasting effects]. / Spasticitetskorrigerande kirurgi kan ge långvariga förbättringar.

Spasticity is a common secondary complication after injuries to the central nervous system (CNS). Spasticity may severely impair arm and hand function, and consequently, affect an individual's function, resulting in disabilities or limiting activities and participation. Today, there is conflicting evidence regarding the effectiveness of available spasticity treatments. The population of adults with spasticity problems due to a CNS injury is heterogenic. A multidisciplinary approach is essential in customizing treatment to individual needs. The planning of treatment should consider the remaining upper limb functions and the patient's potential to improveme. Spasticity-correcting surgery is an option that seems underutilized. This article describes a structured and standardized program for the preoperative assessment, the surgical technique and the postoperative rehabilitation of patients undergoing upper limb spasticity-correcting surgery.

Scapholunate Ligament Reconstruction without Immobilization Is Safe and Leads to Better Functional Results.

Background Reconstruction of the scapholunate ligament (SL) is associated with a reduction in range of motion. In this study, we compared SL reconstruction and early mobilization with SL reconstruction and Kirschner wires (K-wire) fixation for 6 weeks. Methods We performed a nonrandomized trial involving patients with an arthroscopically confirmed SL ligament injury. In total, 11 patients were assigned to SL reconstruction with internal brace augmentation and early mobilization and 10 were assigned to standard SL reconstruction and K-wire fixation for 6 weeks before mobilization started. We assessed the range of motion, grip strength, functional status, intensity of pain, global perceived effect, and duration until return to work. Results In both groups, there was one traumatic breakout of the reconstructed ligament and two patients in the control group were lost to follow-up. The 10 patients in the internal brace group had a mean increase in wrist flexion of 1.8 degrees at 1 year compared with a decrease in wrist flexion of 13.4 degrees in the seven patients of the control group. Wrist extension increased by 4.5 degrees in the internal brace group and decreased by 4.5 degrees in the control group. In addition, the internal brace group scored 6.1 (much improved) for the global perceived effect and the control group 4.7 (slightly improved), and treatment without immobilization resulted in an earlier return to work (35.1 vs. 73.6 days). Conclusions In SL reconstruction, internal brace augmentation and early mobilization result in improved wrist flexion and extension, higher satisfaction, and earlier return to work.

Technique and biomechanical properties of the side-to-side Z-lengthening in spasticity-correcting surgery- a study on porcine tendons.

Spasticity-correcting surgery in patients with injuries to upper motor neurons include various techniques, including tenotomies and lengthening of tendons of spastic muscles. Early mobilization including active loading of a lengthened tendon is essential to prevent adhesion formation, which necessitates a lengthening technique that resists the forces produced by the involved muscles. This study on animal tendons reports the biomechanical properties in regards to elongation and load to failure in porcine tendons lengthened by either a 3 or 5 cm overlap and tested in a simple force rig. The lengthening technique used in these tendons is described in step-by-step detail. The mean elongation of 20 lengthened tendons at 100 N was 10 mm for tendons with a 3 cm overlap and 6 mm for tendons with a 5 cm overlap. The mean peak load at failure of the construct was 138 N for lenghened tendons with a 3 cm overlap and 201 N for tendons with a 5 cm overlap. The results of this study indicate that a tendon lengthened by the described technique with a mere overlap of three cm will withstand the estimated forces elicited by muscles in the forearm immediately after surgery.

[The combination of nerve and tendon transfers offers good possibilities for hand function in patients with cervical spinal cord injury]. / Så kan funktion i hand och arm återfås efter halsryggmärgsskada.

Regaining upper extremity function is a prioritized matter for patients with tetraplegia after a cervical spinal cord injury (cSCI). The purpose of this article is to describe the current evidence and treatment strategies for upper extremity reconstruction after cSCI at the Centre for Advanced Reconstruction of Extremities, Sahlgrenska University Hospital, Sweden. The specialized unit works in a multidisciplinary setting to optimize the care of the patient population. Preoperative planning and an individualized treatment according to the needs and abilities of the patient are considered key points to achieve the best possible outcome. The addition of nerve transfers to the established method of tendon transfers for grip reconstruction has led to increased possibilities to achieve both functional hand opening and grip. Here we present our preferred method of upper extremity reconstruction, which involves a two-staged procedure where the tendon-based grip reconstruction is preceded by nerve transfer of the supinator to posterior intraosseous nerve whenever possible. Important clinical aspects as well as future perspectives are discussed.

Effect of Delayed Cord Clamping on Neurodevelopment at 3 Years: A Randomized Controlled Trial.

INTRODUCTION: Iron deficiency (ID) is associated with poor neurodevelopment. We have previously shown that delayed umbilical cord clamping (CC) improves iron stores at 8 months and neurodevelopment at 1 year in term, healthy infants in Nepal. OBJECTIVE: The aim of this study was to assess the effects of delayed CC (≥180 s) compared to early CC (≤60 s) on neurodevelopment using the Ages and Stages Questionnaire (ASQ) at age 3 years. METHODS: In 2014, 540 healthy Nepalese infants born at term were randomized in a 1:1 ratio to delayed or early CC. At 3 years of age, ASQ assessment was performed by phone interviews with parents. A score >1 standard deviation below the mean was defined as "at risk" for developmental impairment. RESULTS: At 3 years of age, 350 children were followed up, 170 (63.0%) in the early CC group and 180 (66.7%) in the delayed CC group. No significant differences in ASQ scores in any domains between groups were found. However, more girls were "at risk" for affected gross motor development in the early CC group: 14 (18.9%) versus 6 (6.3%), p = 0.02. CONCLUSION: There were no significant differences in ASQ scores in any domains between groups. In the subgroup analysis, fewer girls who underwent delayed CC were "at risk" for delayed gross motor development. Due to the pronounced difference in iron stores at 8 months postpartum in this cohort, follow-up studies at an older age are motivated since neurodevelopmental impairment after early ID may be more detectable with increasing age.

Helicopter Emergency Medical Services Response to a Major Incident.

Major incidents account for a vast number of consequences, whether it be individual morbidity and mortality or economic disruption and expense. Because of the infrequent nature, it poses a variety of unique risks and challenges for individual emergency medical services systems. Air ambulances are usually dispatched based on the clinical presentation of an individual patient who needs emergent critical care intervention. The response to a major incident is unusual and infrequent, but the benefit of tasking air ambulances to such incidents has been described by various authors. Here, such a response is described in a low- to middle-income country that saw the immediate tasking of 2 separate air ambulances to a single, multivehicle collision with multiple injured patients that occurred near a small, rural hospital not capable of treating critically ill patients. The benefits of tasking of the air ambulance in the sense of additional expertise as well as potential other nonclinical benefits are discussed and described here.

Lipid Composition Affects the Efficiency in the Functional Reconstitution of the Cytochrome c Oxidase.

The transmembrane protein cytochrome c oxidase (CcO) is the terminal oxidase in the respiratory chain of many aerobic organisms and catalyzes the reduction of dioxygen to water. This process maintains an electrochemical proton gradient across the membrane hosting the oxidase. CcO is a well-established model enzyme in bioenergetics to study the proton-coupled electron transfer reactions and protonation dynamics involved in these processes. Its catalytic mechanism is subject to ongoing intense research. Previous research, however, was mainly focused on the turnover of oxygen and electrons in CcO, while studies reporting proton turnover rates of CcO, that is the rate of proton uptake by the enzyme, are scarce. Here, we reconstitute CcO from R. sphaeroides into liposomes containing a pH sensitive dye and probe changes of the pH value inside single proteoliposomes using fluorescence microscopy. CcO proton turnover rates are quantified at the single-enzyme level. In addition, we recorded the distribution of the number of functionally reconstituted CcOs across the proteoliposome population. Studies are performed using proteoliposomes made of native lipid sources, such as a crude extract of soybean lipids and the polar lipid extract of E. coli, as well as purified lipid fractions, such as phosphatidylcholine extracted from soybean lipids. It is shown that these lipid compositions have only minor effects on the CcO proton turnover rate, but can have a strong impact on the reconstitution efficiency of functionally active CcOs. In particular, our experiments indicate that efficient functional reconstitution of CcO is strongly promoted by the addition of anionic lipids like phosphatidylglycerol and cardiolipin.

Kinetic advantage of forming respiratory supercomplexes.

Components of respiratory chains in mitochondria and some aerobic bacteria assemble into larger, multiprotein membrane-bound supercomplexes. Here, we address the functional significance of supercomplexes composed of respiratory-chain complexes III and IV. Complex III catalyzes oxidation of quinol and reduction of water-soluble cytochrome c (cyt c), while complex IV catalyzes oxidation of the reduced cyt c and reduction of dioxygen to water. We focus on two questions: (i) under which conditions does diffusion of cyt c become rate limiting for electron transfer between these two complexes? (ii) is there a kinetic advantage of forming a supercomplex composed of complexes III and IV? To answer these questions, we use a theoretical approach and assume that cyt c diffuses in the water phase while complexes III and IV either diffuse independently in the two dimensions of the membrane or form supercomplexes. The analysis shows that the electron flux between complexes III and IV is determined by the equilibration time of cyt c within the volume of the intermembrane space, rather than the cyt c diffusion time constant. Assuming realistic relative concentrations of membrane-bound components and cyt c and that all components diffuse independently, the data indicate that electron transfer between complexes III and IV can become rate limiting. Hence, there is a kinetic advantage of bringing complexes III and IV together in the membrane to form supercomplexes.

Structural changes at the surface of cytochrome c oxidase alter the proton-pumping stoichiometry.

Data from earlier studies showed that minor structural changes at the surface of cytochrome c oxidase, in one of the proton-input pathways (the D pathway), result in dramatically decreased activity and a lower proton-pumping stoichiometry. To further investigate how changes around the D pathway orifice influence functionality of the enzyme, here we modified the nearby C-terminal loop of subunit I of the Rhodobacter sphaeroides cytochrome c oxidase. Removal of 16 residues from this flexible surface loop resulted in a decrease in the proton-pumping stoichiometry to <50% of that of the wild-type enzyme. Replacement of the protonatable residue Glu552, part of the same loop, by an Ala, resulted in a similar decrease in the proton-pumping stoichiometry without loss of the O2-reduction activity or changes in the proton-uptake kinetics. The data show that minor structural changes at the orifice of the D pathway, at a distance of ~40 Šfrom the proton gate of cytochrome c oxidase, may alter the proton-pumping stoichiometry of the enzyme.

Countertransference in Swedish psychotherapists: testing the factor structure of the Therapist Response Questionnaire.

Questionnaires need testing of reliability and factor structure before clinical use or research in new languages or cultures. The aim of this study was to evaluate the Therapist Response Questionnaire (TRQ) in Sweden compared to corresponding factor analyses in USA and Italy. A national sample of psychotherapists (N=242) registered their countertransference with a single client using TRQ. The data were analyzed with confirmatory factor analysis (CFA) to test factor structures from previous studies, and exploratory factor analysis (EFA). The CFA did not verify the factor structure from the previous studies. The EFA extracted seven factors as the best solution: Helpless/Inadequate, Overwhelmed/Disorganized, Hostile/Angry, Parental/Protective, Disengaged, Special/Overinvolved, Sexualized. Analysis of convergent validity indicated that five of these could be considered equivalent to factors in the previous studies, and the remaining two were conceptually related to corresponding factors. Even though the factor structure was not confirmed by the CFA, the concordance was large, indicating a reliable self-report instrument with promising validity for measurement of complex aspects of countertransference. Common countertransference themes can inform psychotherapy supervision and education, give feedback to the therapist, and lay ground for a taxonomy for therapist reactions and feelings.

Trends in tissue engineering for blood vessels.

Over the years, cardiovascular diseases continue to increase and affect not only human health but also the economic stability worldwide. The advancement in tissue engineering is contributing a lot in dealing with this immediate need of alleviating human health. Blood vessel diseases are considered as major cardiovascular health problems. Although blood vessel transplantation is the most convenient treatment, it has been delimited due to scarcity of donors and the patient's conditions. However, tissue-engineered blood vessels are promising alternatives as mode of treatment for blood vessel defects. The purpose of this paper is to show the importance of the advancement on biofabrication technology for treatment of soft tissue defects particularly for vascular tissues. This will also provide an overview and update on the current status of tissue reconstruction especially from autologous stem cells, scaffolds, and scaffold-free cellular transplantable constructs. The discussion of this paper will be focused on the historical view of cardiovascular tissue engineering and stem cell biology. The representative studies featured in this paper are limited within the last decade in order to trace the trend and evolution of techniques for blood vessel tissue engineering.

A single-step competitive binding assay for mapping of single DNA molecules.

Optical mapping of genomic DNA is of relevance for a plethora of applications such as scaffolding for sequencing and detection of structural variations as well as identification of pathogens like bacteria and viruses. For future clinical applications it is desirable to have a fast and robust mapping method based on as few steps as possible. We here demonstrate a single-step method to obtain a DNA barcode that is directly visualized using nanofluidic devices and fluorescence microscopy. Using a mixture of YOYO-1, a bright DNA dye, and netropsin, a natural antibiotic with very high AT specificity, we obtain a DNA map with a fluorescence intensity profile along the DNA that reflects the underlying sequence. The netropsin binds to AT-tetrads and blocks these binding sites from YOYO-1 binding which results in lower fluorescence intensity from AT-rich regions of the DNA. We thus obtain a DNA barcode that is dark in AT-rich regions and bright in GC-rich regions with kilobasepair resolution. We demonstrate the versatility of the method by obtaining a barcode on DNA from the phage T4 that captures its circular permutation and agrees well with its known sequence.

Electric field controlled magnetic anisotropy in a single molecule.

We have measured quantum transport through an individual Fe(4) single-molecule magnet embedded in a three-terminal device geometry. The characteristic zero-field splittings of adjacent charge states and their magnetic field evolution are observed in inelastic tunneling spectroscopy. We demonstrate that the molecule retains its magnetic properties and, moreover, that the magnetic anisotropy is significantly enhanced by reversible electron addition/subtraction controlled with the gate voltage. Single-molecule magnetism can thus be electrically controlled.

Heterologous carotenoid production in Saccharomyces cerevisiae induces the pleiotropic drug resistance stress response.

To obtain insight into the genome-wide transcriptional response of heterologous carotenoid production in Saccharomyces cerevisiae, the transcriptome of two different S. cerevisiae strains overexpressing carotenogenic genes from the yeast Xanthophyllomyces dendrorhous grown in carbon-limited chemostat cultures was analysed. The strains exhibited different absolute carotenoid levels as well as different intermediate profiles. These discrepancies were further sustained by the difference of the transcriptional response exhibited by the two strains. Transcriptome analysis of the strain producing high carotenoid levels resulted in specific induction of genes involved in pleiotropic drug resistance (PDR). These genes encode ABC-type and major facilitator transporters which are reported to be involved in secretion of toxic compounds out of cells. ß-Carotene was found to be secreted when sunflower oil was added to the medium of S. cerevisiae cells producing high levels of carotenoids, which was not observed when added to X. dendrorhous cells. Deletion of pdr10, one of the induced ABC transporters, decreased the transformation efficiency of a plasmid containing carotenogenic genes. The few transformants that were obtained had decreased growth rates and lower carotenoid production levels compared to a pdr5 deletion and a reference strain transformed with the same genes. Our results suggest that production of high amounts of carotenoids in S. cerevisiae leads to membrane stress, in which Pdr10 might play an important role, and a cellular response to secrete carotenoids out of the cell.

Progressive multifocal leukoencephalopathy after natalizumab monotherapy.

We describe progressive multifocal leukoencephalopathy (PML) caused by infection with human polyomavirus JC virus in a patient with multiple sclerosis who was treated with natalizumab. The first PML symptoms appeared after 14 monthly infusions of the drug. Magnetic resonance imaging (MRI) showed a presumed multiple sclerosis lesion, and JC virus DNA was not detected on polymerase-chain-reaction (PCR) assay of cerebrospinal fluid. The patient's symptoms worsened, and the diagnosis of PML was established with a more sensitive quantitative PCR assay after 16 infusions of natalizumab. Plasma exchange was used to accelerate clearance of natalizumab. Approximately 3 weeks after plasma exchange, an immune-reconstitution inflammatory syndrome appeared. JC virus DNA was no longer detectable on quantitative PCR assay, and the patient's symptoms improved.

Genes involved in carotene synthesis and mating in Blakeslea trispora.

Mating of Blakeslea trispora and other molds of the order Mucorales requires the interaction of mycelia of opposite sex, (+) and (-), leading to the development of specialized structures and to an enhanced accumulation of beta-carotene. Industry obtains beta-carotene by co-cultivating appropriate strains of Blakeslea ("mated cultures"). Gene transcription in single and mated cultures was assayed by cDNA-AFLP, a technique to observe the differential expression of subsets of mRNA fragments. Overexpression in mated cultures is about ten times more frequent than underexpression. We obtained and sequenced fragments of 97 candidate genes that appeared to be overexpressed during mating and confirmed four of them by reverse transcription and real-time PCR. Comparisons with gene sequences from other organisms suggest functions in carotene biosynthesis (4 genes), energy metabolism (8), cell wall synthesis (1), transfer of acetyl groups (1), and regulatory processes (10). Sodium acetate inhibited sexual overexpression in about two-thirds of the candidate genes and acted as a signal with broad effects on the metabolism and the morphology of mated cultures. Our work offers new materials for the study of carotene biosynthesis and its regulation and for the improvement of carotene production with Mucorales.

High-level production of beta-carotene in Saccharomyces cerevisiae by successive transformation with carotenogenic genes from Xanthophyllomyces dendrorhous.

To determine whether Saccharomyces cerevisiae can serve as a host for efficient carotenoid and especially beta-carotene production, carotenogenic genes from the carotenoid-producing yeast Xanthophyllomyces dendrorhous were introduced and overexpressed in S. cerevisiae. Because overexpression of these genes from an episomal expression vector resulted in unstable strains, the genes were integrated into genomic DNA to yield stable, carotenoid-producing S. cerevisiae cells. Furthermore, carotenoid production levels were higher in strains containing integrated carotenogenic genes. Overexpression of crtYB (which encodes a bifunctional phytoene synthase and lycopene cyclase) and crtI (phytoene desaturase) from X. dendrorhous was sufficient to enable carotenoid production. Carotenoid production levels were increased by additional overexpression of a homologous geranylgeranyl diphosphate (GGPP) synthase from S. cerevisiae that is encoded by BTS1. Combined overexpression of crtE (heterologous GGPP synthase) from X. dendrorhous with crtYB and crtI and introduction of an additional copy of a truncated 3-hydroxy-3-methylglutaryl-coenzyme A reductase gene (tHMG1) into carotenoid-producing cells resulted in a successive increase in carotenoid production levels. The strains mentioned produced high levels of intermediates of the carotenogenic pathway and comparable low levels of the preferred end product beta-carotene, as determined by high-performance liquid chromatography. We finally succeeded in constructing an S. cerevisiae strain capable of producing high levels of beta-carotene, up to 5.9 mg/g (dry weight), which was accomplished by the introduction of an additional copy of crtI and tHMG1 into carotenoid-producing yeast cells. This transformant is promising for further development toward the biotechnological production of beta-carotene by S. cerevisiae.

A new group of exo-acting family 28 glycoside hydrolases of Aspergillus niger that are involved in pectin degradation.

The fungus Aspergillus niger is an industrial producer of pectin-degrading enzymes. The recent solving of the genomic sequence of A. niger allowed an inventory of the entire genome of the fungus for potential carbohydrate-degrading enzymes. By applying bioinformatics tools, 12 new genes, putatively encoding family 28 glycoside hydrolases, were identified. Seven of the newly discovered genes form a new gene group, which we show to encode exoacting pectinolytic glycoside hydrolases. This group includes four exo-polygalacturonan hydrolases (PGAX, PGXA, PGXB and PGXC) and three putative exo-rhamnogalacturonan hydrolases (RGXA, RGXB and RGXC). Biochemical identification using polygalacturonic acid and xylogalacturonan as substrates demonstrated that indeed PGXB and PGXC act as exo-polygalacturonases, whereas PGXA acts as an exo-xylogalacturonan hydrolase. The expression levels of all 21 genes were assessed by microarray analysis. The results from the present study demonstrate that exo-acting glycoside hydrolases play a prominent role in pectin degradation.

Cloning of the astaxanthin synthase gene from Xanthophyllomyces dendrorhous (Phaffia rhodozyma) and its assignment as a beta-carotene 3-hydroxylase/4-ketolase.

A gene has been cloned from Xanthophyllomyces dendrorhous by complementation of astaxanthin formation in a beta-carotene accumulating mutant. It consists of 3,166 bp and contains 17 introns. For the beta-carotene mutant ATCC 96815, a single point mutation in the splicing sequence of intron 8 was found. The resulting improper splicing of the mRNA results in an inactive protein. The cDNA of this beta-carotene oxygenase encodes a cytochrome P450 monooxygenase belonging to the 3A subfamily. P450-specific domains were identified including a cytochrome P450 and an oxygen binding motif. Electrons are provided by a cytochrome P450 reductase. Functional characterization of the enzyme by genetic modification of X. dendrorhous demonstrated that this P450 monooxygenase is multifunctional catalyzing all steps from beta-carotene to astaxanthin formation by oxygenation of carbon 3 and 4. The reaction sequence is first 4-ketolation of beta-carotene followed by 3-hydroxylation. A hydroxylation mechanism at allylic carbon atoms has been proposed for the generation of 4-keto and 3-hydroxy groups at both beta-ionone ends.