Immunolocalization of VEGF/VEGFR system in human fetal vomeronasal organ during early development

The vomeronasal system (VNS) is an accessory olfactory structure present in most mammals adhibited to the detection of specific chemosignals implied in social and reproductive behavior. The VNS comprises the vomeronasal organ (VNO), vomeronasal nerve and accessory olfactory bulb. VNO is characterized by a neuroepithelium constituted by bipolar neurons and supporting and stem/progenitor cells. In humans, VNO is present during fetal life and is supposed to possess chemoreceptor activity and participate in gonadotropin-releasing hormone neuronal precursor migration toward the hypothalamus. Instead, the existence and functions of VNO in postnatal life is debated. Vascular endothelial growth factor (VEGF) and its receptors (VEGFRs) have been demonstrated to play fundamental roles in various neurogenic events. However, there are no data regarding the localization and possible function of VEGF/VEGFRs in human fetal VNO. Therefore, this study was conceived to investigate the expression of VEGF/VEGFRs in human VNO in an early developmental period (9–12 weeks of gestation), when this organ appears well structured. Coronal sections of maxillofacial specimens were subjected to peroxidase-based immunohistochemistry for VEGF, VEGFR-1 and VEGFR-2. Double immunofluorescence for VEGF, VEGFR-1 or VEGFR-2 and the neuronal marker protein gene product 9.5 (PGP 9.5) was also performed. VEGF expression was evident in the entire VNO epithelium, with particularly strong reactivity in the middle layer. Strongly VEGF-immunostained cells with aspect similar to bipolar neurons and/or their presumable precursors were detected in the middle and basal layers. Cells detaching from the basal epithelial layer and detached cell groups in the surrounding lamina propria showed moderate/strong VEGF expression. The strongest VEGFR-1 and VEGFR-2 expression was detected in the apical epithelial layer. Cells with aspect similar to bipolar neurons and/or their presumable precursors located in the middle and basal layers and the detaching/detached cells displayed a VEGFR-1 and VEGFR-2 reactivity similar to that of VEGF. The basal epithelial layer exhibited stronger staining for VEGFRs than for VEGF. Cells with morphology and VEGF/VEGFR expression similar to those of the detaching/detached cells were also detected in the middle and basal VNO epithelial layers. Double immunofluorescence using anti-PGP 9.5 antibodies demonstrated that most of the VEGF/VEGFR-immunoreactive cells were neuronal cells. Collectively, our findings suggest that during early fetal development the VEGF/VEGFR system might be involved in the presumptive VNO chemoreceptor activity and neuronal precursor migration.     

Tango knock-ins visualize endogenous activity of G protein-coupled receptors in Drosophila

G protein-coupled receptors (GPCRs) represent a family of seven-pass transmembrane protein receptors whose ligands include neuropeptides and small-molecule neuromodulators such as dopamine and serotonin. These neurotransmitters act at long distances and are proposed to define the ground state of the nervous system. The Drosophila genome encodes approximately 50 neuropeptides and their functions in physiology and behavior are now under intensive studies. Key information currently lacking in the field is the spatiotemporal activation patterns of endogenous GPCRs. Here we report application of the Tango system, a reporter assay to detect GPCR activity, to endogenous GPCRs in the fly genome. We developed a method to integrate the sensor component of the Tango system to the C-terminus of endogenous genes by using genome editing techniques. We demonstrate that Tango sensors in the Sex-peptide receptor (SPR) locus allow sensitive detection of mating-dependent SPR activity in the female reproductive organ. The method is easily applicable to any GPCR and will provide a way to systematically characterize GPCRs in the fly brain.     

Are rats more human than mice?

In contrast to rats, mouse models are nowadays generally used for the investigation of immune responses and immune-mediated diseases, there are many different strains and mouse-specific tools available, and it is easy to generate transgenic and constitutive or inducible knockout mice for any gene. Many immune markers and mechanisms have been detected in mice and have been introduced as gold standard in immunology, however, some turned out to be not unconditionally transferable to the human immune system.Rats have been used more frequently in former days but are mostly outstripped by mice due to the fact that fewer strains are available, they need more space than mice, are more expensive to maintain and breed, and it is extremely difficult to generate transgenic or ko-rats. Consequently, the choice of rat-specific diagnostic tools like antibodies is quite poor and most researchers have switched to mouse models for the investigation of immune mechanisms, while rats are still widely used for toxicology by the pharmaceutical industry. However, it should be taken into consideration that there are some immunological similarities between rats and humans that are not presented in mice. Some of them like MHC class II and Foxp3 expression by activated effector T cells we have detected during our research on the immune response of rat models of experimental autoimmune uveitis.     

The midterm results of a cohort study of patellofemoral arthroplasty from a non-designer centre using an asymmetric trochlear prosthesis

PurposePatellofemoral arthroplasty (PFA) prosthesis with asymmetric trochlear component was introduced as an improvement from existing designs for surgical treatment of symptomatic isolated patellofemoral arthritis. The purpose of this study was to evaluate midterm results in patients who underwent PFA procedure using such prosthesis.MethodsOur study involved a continuous retrospective cohort of patients who underwent PFA using Journey PFA prosthesis with an asymmetric trochlear component, performed between June 2007 and July 2016 at a non-designer centre. The Patient Reported Outcome Measures and patient satisfaction questionnaires were collected for final evaluation.ResultsA total of 103 PFA performed on 79 patients were evaluated. Median age at the time of surgery was 58 years (range 42 to 78 years); the mean follow-up period was 6 years (range 2 to 11 years). Four knees were revised to Total Knee Arthroplasty for reasons not related to the implant. The cumulative survival estimated by the Kaplan–Meier method was 94.3% (95% confidence interval: 88.4%–100%). There were statistically significant improvements in functional outcome scores.ConclusionThis series of patients who underwent PFA with the asymmetric trochlear component has shown promising mid-term results with no implant-related complications.     

Assessment of objective dynamic knee joint control in anterior cruciate ligament deficient and reconstructed individuals

BackgroundThere is a lack of objective dynamic knee joint control measures that can be related to the status of the anterior cruciate ligament (ACL). The purpose of this study was to introduce two novel measures and apply a third to determine how dynamic knee joint control changes in relation to ACL status during dynamic movements.MethodsTwenty patients (13 male) were tested pre- (ACLd) and 10-months post- (ACLr) ACL reconstructive surgery and matched to an uninjured participant (CON). Kinetic and kinematic data were synchronously recorded with a force platform and motion capture system. Three objective control measures including dynamic angular stiffness, knee joint center excursion (KJCE), and knee joint center boundary (KJCB) were assessed for each participant when completing the side cut and hop tasks.ResultsDuring the side cut, stiffness was found to be significantly lower in ACLd (0.06 ± 0.01 Nm/kg/°) and ACLr (0.07 ± 0.02 Nm/kg/°) compared to CON (0.08 ± 0.02 Nm/kg/°), while there were no differences in stiffness during the hop. No significant differences were observed in the KJCE during the side cut, while KJCE was significantly greater (p = 0.006) during the hop in CON compared to the ACLd. There were no differences in KJCB.ConclusionsThese high-functioning ACL injured in both ACLd and ACLr phases, aside from reduced stiffness, were able to complete both tasks with similar dynamic control as the CON. Although improvements in self-perceived control between ACLd and ACLr have been observed, this lack of improvement in objective control demonstrates a gap between a patient's self-efficacy and the level of control.     

微创手术机器人机械臂结构设计与工作空间分析

目的现有双平行四边形机构设计的机械臂需要串联一个伸缩自由度机构,导致机械臂体积增大,运动灵活性降低,持镜臂与持器械臂发生干涉。针对这一问题,提出一种新型机械臂结构,增强运动灵活性、减小机械臂的体积。方法采用5连杆机构、滑块与滑轨的低副机构和丝传动机构设计机械臂,实现末端执行器的伸缩运动,建立机械臂的运动学模型,以MATLAB为仿真工具验证零位关节角D-H参数的正确性,求解出机械臂的运动方程;同时运用蒙特卡洛算法,得到末端执行器的三维工作空间范围,并对3条机械臂作术前动物实验规划;最后,在猪体内进行胆囊切除等操作,验证机械臂的双5连杆2自由度设计的合理性和可操作性。结果蒙特卡洛算法在MATLAB环境下得到的工作空间范围为:-650.4 mmx649 mm,163.8 mmy1 202 mm,-254.6 mmz829.8 mm,并成功完成16例猪的胆囊切除实验,平均操作时间为51 min。结论新型设计的双5连杆2自由度机械臂在猪体内成功完成胆囊切除等操作,且猪术后无其他不良症状,机械臂之间的运动无干涉,充分验证了所提出的微创手术机器人机械臂设计方案的可行性。