A novel in vitro co-culture model to examine contact formation between astrocytic processes and cerebral vessels

Here, we developed a novel in vitro co-culture model, in which process-bearing astrocytes and isolated cerebral microvessels from mice were co-cultured. Astrocytes formed contacts with microvessels from both adult and neonatal mice. However, concentrated localization of the immunofluorescence signal for aquaporin-4 (AQP4) at contact sites between perivascular endfoot processes and blood vessels was only detected with neonatal mouse microvessels. Contact between astrocytic processes and microvessels was retained, whereas concentrated localization of AQP4 signal at contact sites was lost, by knockdown of dystroglycan or α-syntrophin, reflecting polarized localization of AQP4 at perivascular regions in the brain. Further, using our in vitro co-culture model, we found that astrocytes predominantly extend processes to pericytes located at the abluminal surface of microvessels, providing additional evidence that this model is representative of the in vivo situation. Altogether, we have developed a novel in vitro co-culture model that can reproduce aspects of the in vivo situation and is useful for assessing contact formation between astrocytes and blood vessels.     

Sensor-to-body calibration procedure for clinical motion analysis of lower limb using magnetic and inertial measurement units

Magnetic and Inertial measurement units (MIMUs) have become exceedingly popular for ambulatory human motion analysis during the past two decades. However, measuring anatomically meaningful segment and joint kinematics requires virtual alignment of the MIMU frame with the anatomical frame of its corresponding segment. Therefore, this paper presents a simple calibration procedure, based on MIMU readouts, to align the inertial frame of the MIMU with the anatomical frames, as recommended by ISB. The proposed calibration includes five seconds of quiet standing in a neutral posture followed by ten consecutive hip flexions/extensions. This procedure will independently calibrate MIMUs attached to the pelvis, thigh, shank, and foot. The accuracy and repeatability of the calibration procedure and the 3D joint angle estimation were validated against the gold standard motion capture system by an experimental study with ten able-bodied participants. The procedure showed high test-retest repeatability in aligning the MIMU frame with its corresponding anatomical frame, i.e., the helical angle between the MIMU and anatomical frames did not significantly differ between the test and retest sessions (except for thigh MIMU). Compared to previously introduced procedures, this procedure attained the highest inter-participant repeatability (inter-participant coefficient of variations of the helical angle: 20.5–42.2%). Further, the proposed calibration would reduce the offset errors of the 3D joint angle estimation (up to 12.8 degrees on average) compared to joint angle estimation without calibration (up to 26.3 degrees on average). The proposed calibration enables MIMU to measure clinically meaningful gait kinematics.     

Stance and swing phase knee flexion recover at different rates following total knee arthroplasty: An inertial measurement unit study

Total knee arthroplasty (TKA) is the most common joint replacement in the United States. Range of motion (ROM) monitoring includes idealized clinic measures (e.g. goniometry during passive ROM) that may not accurately represent knee function. Accordingly, a novel, portable, inertial measurement unit (IMU) based ROM measurement method was developed, validated, and implemented. Knee flexion was computed via relative motion between two IMUs and validated via optical motion capture (p > 0.05). Prospective analyses of 10 healthy individuals (5M, 50 ± 19 years) and 20 patients undergoing TKA (3 lost to follow up, 10M, 65 ± 6 years) were completed. Controls wore IMUs for 1-week. Patients wore IMUs for 1-week pre-TKA, 6-weeks immediately post-TKA, and 1-week at 1-year post-TKA. Flexion was computed continuously each day (8–12 h). Metrics included daily maximum flexion and flexion during stance/swing phases of gait. Maximum flexion was equal between cohorts at all time points. Contrastingly, patient stance and swing flexion were reduced pre-TKA, yet improved post-TKA. Specifically, patient stance and swing flexion were reduced below control/pre-TKA values during post-TKA week 1. Stance flexion exceeded pre-TKA and equaled control levels after week 2. However, swing flexion only exceeded pre-TKA and equaled control levels at 1-year post-TKA. This novel method improves upon the accuracy/portability of current methods (e.g. goniometry). Interestingly, surgery did not impact maximum ROM, yet improved the ability to flex during gait allowing more efficient and safe ambulation. This is the first study continuously monitoring long-term flexion before/after TKA. The results offer richer information than clinical measures about expected TKA rehabilitation.     

Delayed inflammation decrease is associated with mortality in Tocilizumab-treated critically ill SARS-CoV-2 patients: A retrospective matched-cohort analysis

Little is known about the immuno-inflammatory response to Tocilizumab and its association with outcome in critically-ill SARS-CoV2 pneumonia. In this multicenter retrospective cohort of SARS-CoV-2 patients admitted to three intensive care units between March and April 2020, we matched on gender and SAPS II 21 Tocilizumab-treated patients to 42 non-treated patients. Need for mechanical ventilation was 76% versus 79%. IL-6, C-reactive protein, and fibrinogen had been collected within the first days of admission (T1), 3 d (T2) and 7 d (T3) later. Tocilizumab-treated patients had persistently higher IL-6 plasma levels and persistently lower C-Reactive protein and fibrinogen levels. Among Tocilizumab-treated patients, baseline levels of inflammatory biomarkers were not different according to outcome. Conversely, C-reactive protein and fibrinogen decrease was delayed in non-survivors. C-Reactive protein decreased at T1 in survivors (45 [30–98] vs 170 [69–204] mg/l, P < 0.001) but only at T2 in non-survivors (37 [13–74] vs 277 [235–288], P = 0.03). Fibrinogen decreased at T2 in survivors (4.11 [3.58–4.69] vs 614 [5.61–7.85] g/l, P = 0.005) but not in non-survivors (4.79 [4.12–7.58] vs 7.24 [6.22–9.24] g/l, P = 0.125). Tocilizumab treatment was thus associated with a persistent both increase in plasma IL-6, and decrease in C-reactive protein and fibrinogen. Among Tocilizumab-treated patients, the decrease in inflammatory biomarkers was delayed in non-survivors.     

综合性医院重症监护病房病原菌分离情况分析

目的探讨重症监护病房（Icu）医院内感染的临床特点及病原菌种类、分布情况,为临床合理使用抗菌药物、预防和控制医院感染提供参考和依据。方法采用前瞻性监测与回顾性调查相结合的方法,对ICU患者的临床资料进行统计分析。结果ICU病人标本中分离出病原菌593株,得出菌种分布与感染情况。结论重症监护病房医院内感染发生率高,以呼吸道感染为主,主要病原菌以革兰阴性非发酵菌为主,加强ICU患者感染的控制,可减少ICU医院内感染的发生。     

Ribosomal DNA Intergenic Spacer of the Swimming Crab, Charybdis japonica

We have determined the full sequence of the ribosomal DNA intergenic spacer (IGS) of the swimming crab, Charybdis japonica, by long PCR for the first time in crustacean decapods. The IGS is 5376 bp long and contains two nonrepetitive regions separated by one long repetitive region, which is composed mainly of four subrepeats (subrepeats I, II, III, and IV). Subrepeat I contains nine copies of a 60-bp repeat unit, in which two similar repeat types (60 bp-a and 60 bp-b) occur alternatively. Subrepeat II consists of nine successive repeat units with a consensus sequence length of 142 bp. Subrepeat III consists of seven copies of another 60-bp repeat unit (60 bp-c) whose sequence is complementary to that of subrepeat I. Immediately downstream of subrepeat III is subrepeat IV, consisting of three copies of a 391-bp repeat unit. Based on comparative analysis among the subrepeats and repeat units, a possible evolutionary process responsible for the formation of the repetitive region is inferred, which involves the duplication of a 60-bp subrepeat unit (60 bp-c) as a prototype. Received: 13 April 1999 / Accepted: 2 August 1999     

去甲肾上腺素介导低氧引起家兔颈动脉体神经电活动增加

在30只家兔颈动脉体-窦神经(CSN)标本上, 记录了窦神经中39个对低氧反应敏感的化学感受性单位由去甲肾上腺素(NA)及其拮抗剂引起的反应。结果如下 (1)以低氧的改良台氏液(MTS)灌流标本时, 19个单位放电频率由0.13±0.06增至0.25±0.12 imp/s (P<0.001); (2)在灌流液中加入去甲肾上腺素(10     

Population Genetic Structure and Conservation of the Lesser White-Fronted GooseAnser erythropus

The lesser white-fronted goose is a sub-Arctic species with a currently fragmented breeding range, which extends from Fennoscandia to easternmost Siberia. The population started to decline at the beginning of the last century and, with a current world population estimate of 25,000 individuals, it is the most threatened of the Palearctic goose species. Of these, only 30–50 pairs breed in Fennoscandia. A fragment of the control region of mtDNA was sequenced from 110 individuals from four breeding, one staging and two wintering areas to study geographic subdivisions and gene flow. Sequences defined 15 mtDNA haplotypes that were assigned to two mtDNA lineages. Both the mtDNA lineages were found from all sampled localities indicating a common ancestry and/or some level of gene flow. Analyses of molecular variance showed significant structuring among populations ( _ST 0.220, P < 0.001). The results presented here together with ecological data indicate that the lesser white-fronted goose is fragmented into three distinctive subpopulations, and thus, the conservation status of the species should be reconsidered.     

Bracketed generic inactivation of rodent retroviruses by low pH treatment for monoclonal antibodies and recombinant proteins

Viral safety is a predominant concern for monoclonal antibodies (mAbs) and other recombinant proteins (RPs) with pharmaceutical applications. Certain commercial purification modules, such as nanofiltration and low-pH inactivation, have been observed to reliably clear greater than 4 log(10) of large enveloped viruses, including endogenous retrovirus. The concept of "bracketed generic clearance" has been proposed for these steps if it could be prospectively demonstrated that viral log(10) reduction value (LRV) is not impacted by operating parameters that can vary, within a reasonable range, between commercial processes. In the case of low-pH inactivation, a common step in mAb purification processes employed after protein A affinity chromatography, these parameters would include pH, time and temperature of incubation, the content of salts, protein concentration, aggregates, impurities, model protein pI, and buffer composition. In this report, we define bracketed generic clearance conditions, using a prospectively defined bracket/matrix approach, where low-pH inactivation consistently achieves >or=4.6 log(10) clearance of xenotropic murine leukemia virus (X-MLV), a model for rodent endogenous retrovirus. The mechanism of retrovirus inactivation by low-pH treatment was also investigated.     

Biosynthesis of O-antigens: genes and pathways involved in nucleotide sugar precursor synthesis and O-antigen assembly

The O-antigen is an important component of the outer membrane of Gram-negative bacteria. It is a repeat unit polysaccharide and consists of a number of repeats of an oligosaccharide, the O-unit, which generally has between two and six sugar residues. O-Antigens are extremely variable, the variation lying in the nature, order and linkage of the different sugars within the polysaccharide. The genes involved in O-antigen biosynthesis are generally found on the chromosome as an O-antigen gene cluster, and the structural variation of O-antigens is mirrored by genetic variation seen in these clusters. The genes within the cluster fall into three major groups. The first group is involved in nucleotide sugar biosynthesis. These genes are often found together in the cluster and have a high level of identity. The genes coding for a significant number of nucleotide sugar biosynthesis pathways have been identified and these pathways seem to be conserved in different O-antigen clusters and across a wide range of species. The second group, the glycosyl transferases, is involved in sugar transfer. They are often dispersed throughout the cluster and have low levels of similarity. The third group is the O-antigen processing genes. This review is a summary of the current knowledge on these three groups of genes that comprise the O-antigen gene clusters, focusing on the most extensively studied E. coli and S. enterica gene clusters.