A safe insect-based Chikungunya fever vaccine affords rapid and durable protection in cynomolgus macaques.

Chikungunya virus (CHIKV), which induces chikungunya fever and chronic arthralgia, is an emerging public health concern. Safe and efficient vaccination strategies are needed to prevent or mitigate virus-associated acute and chronic morbidities for preparation of future outbreaks. Eilat (EILV)/CHIKV, a chimeric alphavirus which contains the structural proteins of CHIKV and the non-structural proteins of EILV, does not replicate in vertebrate cells. The chimeric virus was previously reported to induce protective adaptive immunity in mice. Here, we assessed the capacity of the virus to induce quick and durable protection in cynomolgus macaques. EILV/CHIKV protected macaques from wild-type (WT) CHIKV infection one year after a single dose vaccination. Transcriptome and in vitro functional analyses reveal that the chimeric virus triggered toll-like receptor signaling and T cell, memory B cell and antibody responses in a dose-dependent manner. Notably, EILV/CHIKV preferentially induced more durable, robust, and broader repertoire of CHIKV-specific T cell responses, compared to a live attenuated CHIKV 181/25 vaccine strain. The insect-based chimeric virus did not cause skin hypersensitivity reactions in guinea pigs sensitized to mosquito bites. Furthermore, EILV/CHIKV induced strong neutralization antibodies and protected cynomolgus macaques from WT CHIKV infection within six days post vaccination. Transcriptome analysis also suggest that the chimeric virus induction of multiple innate immune pathways, including Toll-like receptor signaling, type I IFN and IL-12 signaling, antigen presenting cell activation, and NK receptor signaling. Our findings suggest that EILV/CHIKV is a safe, highly efficacious vaccine, and provides both rapid and long-lasting protection in cynomolgus macaques.

Do strength of zirconia-abutment-interfaces depend on cement, zirconia type and titanium abutment dimensions?

OBJECTIVES: Part 1 of this study investigates the influence of zirconia types, chimney heights, and gingival heights on the strength of the zirconia-abutment-interface. Part 2 extends the analysis to include adhesive brands and macro-retentions. METHODS: In Part 1, the study utilized three zirconia types (700 MPa, 1000 MPa, 1200 MPa) to fabricate 234 screw-retained zirconia crowns with varying chimney heights (3.5 mm, 4.1 mm, 5 mm) and gingival heights (0.65 mm, 1.2 mm, 3 mm) of the titanium abutments. All adherend surfaces underwent sandblasting with aluminum oxide before cementation with a specific resin cement. In Part 2, the investigation of 240 screw-retained zirconia crowns focused on a single zirconia type (1000 MPa) with chimney heights of 3.5 mm and 5 mm and a gingival height of 0.65 mm of the titanium abutments, cemented with three different resin cements. All adherent surfaces underwent sandblasting with aluminum oxide before cementation, whereas 120 out of 240 abutments received additional macro retentions. Storage in water at 37 °C for 24 h preceded the tensile test. RESULTS: The study revealed a substantial impact of chimney height and zirconia type on the bond strength of the zirconia-abutment-interface. Neither adhesive brands nor macro retentions significantly impacted the bond strength. Fracture incidence was significantly influenced by gingival height and zirconia type in part 1, whereas in part 2 smaller chimney heights correlated with a higher fracture incidence. SIGNIFICANCE: This study contributes insights into the complex interplay of factors influencing the zirconia-abutment-interface. The results provide a foundation for refining clinical approaches, emphasizing the importance of chimney height and zirconia type in achieving successful anterior gap implant restorations.

Can we accelerate rehabilitation following reverse shoulder arthroplasty? A systematic review.

Background: There is no consensus concerning the rehabilitation protocol following reverse shoulder arthroplasty. Several patients are expecting to be able to use their arms for sports or recreation shortly after their operation. Methods: This review was designed as an intervention systematic review with narrative analysis. Authors searched English literature in PubMed and Embase databases from 1/1/1989 until July 2022. Controlled studies comparing rehabilitation protocols for patients undergoing reverse shoulder arthroplasty were included. Data quality was examined with the Cochrane risk of a bias assessment tool for randomized trials, the Methodological Index for Non-Randomized studies (MINORS) tool, as well as the Grading of Recommendations Assessment Development and Evaluation (GRADE) approach. Results: Three studies were finally analyzed. At 3 months post-op, forward flexion was found to be significantly higher in the early rehabilitation group (140.5, 95% confidence intervals (CIs): 135.10-145.89; the delayed rehabilitation group mean was 131.24, 95% CI: 125.73-136.74; p = 0.019). Twelve months post-op, no significant difference in any clinical or patient-reported outcome was shown. More complications were reported in the 6 weeks-delayed rehabilitation group. Discussion: Newer regimes permit immediate shoulder mobilization but may not be applied to every patient. The lack of strong evidence warrants the need for future controlled studies; subsequently, postoperative rehabilitation should be individualized.

Influence of cuff stiffness on hemodynamics and perceived cuff pressure in the upper extremities in males and females: implications for practical blood flow restriction training.

BACKGROUND: Practical blood flow restriction (pBFR) during exercise is a cost-saving alternative to traditional blood flow restriction using pneumatic cuffs, particularly when exercising in a group setting. Depending on the pBFR technique, several factors (e.g., cuff width, limb circumference) have already been shown to be of importance when applying the pBFR pressure. Given that elastic cuffs are often used for pBFR, the cuff stiffness might be an additional influencing factor. Therefore, the present study compared the acute effects of three elastic cuffs with identical width but different stiffness (high stiffness (HS), medium stiffness (MS), and low stiffness (LS)) on hemodynamic measures and perceived cuff pressure at rest. METHODS: In a randomized, counter-balanced cross-over study, 36 young and normotensive participants completed three experimental trials. After a 10-min rest period in supine position, the cuff was loosely and proximally applied to the right upper arm. Following baseline data recording, the cuff was successively tightened in 10%-increments with respect to the limb circumference (%overlap) until arterial blood flow was occluded. At baseline and during each %overlap, systolic peak blood flow velocity of the brachial artery, rating of perceived cuff pressure, as well as muscle oxygen saturation and total hemoglobin concentration of the biceps brachii muscle were recorded. RESULTS: The %overlap required to occlude arterial blood flow was different between the three cuffs (HS: 30.9 ± 3.8%, MS: 43.9 ± 6.1%, LS: 54.5 ± 8.3%). Furthermore, at 30% overlap, systolic peak blood flow velocity was lower when applying the HS (9.0 ± 10.9 cm∙s- 1) compared to MS (48.9 ± 21.9 cm∙s- 1) and LS cuff (62.9 ± 19.1 cm∙s- 1). Rating of perceived cuff pressure at 30% overlap was higher when using the HS (6.5 ± 1.5 arbitrary unit (a.u.)) compared to MS (5.1 ± 1.4 a.u.) and LS cuff (4.9 ± 1.5 a.u.) with no difference between the MS and LS cuff. However, muscle oxygen saturation and total hemoglobin concentration were not different between the three cuffs. CONCLUSIONS: The present study revealed that the cuff stiffness influenced blood flow velocity and arterial occlusion pressure. Therefore, cuff stiffness seems an important factor for the application of pBFR.

Detection of La Crosse Virus In Situ and in Individual Progeny to Assess the Vertical Transmission Potential in Aedes albopictus and Aedes aegypti.

La Crosse virus (LACV) is circulating in the midwestern and southeastern states of the United States and can cause human encephalitis. The main vector of the virus is the eastern tree-hole mosquito, Aedes triseriatus. Ae. albopictus has been also described as a natural LACV vector, while Ae. aegypti has been infected with the virus under laboratory conditions. Here, we compare the vertical transmission potential of LACV in Ae. albopictus and Ae. aegypti, with emphasis given to the ovarian infection patterns that the virus generates in both species. Both mosquito species received artificial bloodmeals containing LACV. At defined time points post-infection/bloodmeal, midguts, head tissue, and ovaries were analyzed for the presence of virus. Viral infection patterns in the ovaries were visualized via immunofluorescence confocal microscopy and immunohistopathology assays using an LACV-specific monoclonal antibody. In Ae. aegypti, LACV was confronted with midgut infection and escape barriers, which were much less pronounced in Ae. albopictus, resulting in a significantly higher prevalence of infection in the latter. Following the ingestion of a single virus-containing bloodmeal, no progeny larvae were found to be virus-infected. Regardless, females of both species showed the presence of LACV antigen in their ovariole sheaths. Furthermore, in a single Ae. albopictus female, viral antigen was associated with the nurse cells inside the primary follicles. Following the ingestion of a second non-infectious bloodmeal at 7- or 10-days post-ingestion of an LACV-containing bloodmeal, more progeny larvae of Ae. albopictus than of Ae. aegypti were virus-infected. LACV antigen was detected in the egg chambers and ovariole sheaths of both mosquito species. Traces of viral antigen were also detected in a few oocytes from Ae. albopictus. The low level of vertical transmission and the majority of the ovarian infection patterns suggested the transovum rather than transovarial transmission (TOT) of the virus in both vector species. However, based on the detection of LACV antigen in follicular tissue and oocytes, there was the potential for TOT among several Ae. albopictus females. Thus, TOT is not a general feature of LACV infection in mosquitoes. Instead, the TOT of LACV seems to be dependent on its particular interaction with the reproductive tissues of a female.

Transgene-induced cell death following dengue-2 virus infection in Aedes aegypti.

Dengue viruses (DENVs) are mosquito-borne flaviviruses causing millions of human infections each year and pose a challenge for public health systems worldwide. Aedes aegypti is the principal vector species transmitting DENVs to humans. Controlling Ae. aegypti is difficult due to the abundance of breeding sites and increasing insecticide resistance in the vector populations. Developing new vector control strategies is critical for decreasing the disease burden. One potential approach is genetically replacing Ae. aegypti populations with vector populations highly resistant to DENV transmission. Here, we focus on an alternative strategy for generating dengue 2 virus (DENV-2) resistance in genetically-modified Ae. aegypti in which the mosquitoes express an inactive form of Michelob_x (Mx), an antagonist of the Inhibitor of Apoptosis (IAP), to induce apoptosis in those cells in which actively replicating DENV-2 is present. The inactive form of Mx was flanked by the RRRRSAG cleavage motif, which was recognized by the NS2B/NS3 protease of the infecting DENV-2 thereby releasing and activating Mx which then induced apoptosis. Our transgenic strain exhibited a significantly higher mortality rate than the non-transgenic control when infected with DENV-2. We also transfected a DNA construct containing inactive Mx fused to eGFP into C6/36 mosquito cells and indirectly observed Mx activation on days 3 and 6 post-DENV-2 infections. There were clear signs that the viral NS2B/NS3 protease cleaved the transgene, thereby releasing Mx protein into the cytoplasm, as was confirmed by the detection of eGFP expression in infected cells. The present study represents proof of the concept that virus infection can be used to induce apoptosis in infected mosquito cells.

[Blood flow restriction training as a treatment option for lateral elbow tendinopathy-a study presentation]. / Blood-flow-restriction-Training als Behandlungsoption der Epicondylopathia humeri radialis ­ eine Studienvorstellung.

Blood flow restriction training, developed in 1966 in Japan, is a training modality that utilizes partial arterial and complete venous blood flow occlusion. Combined with low load resistance training, it aims to induce hypertrophy and strength gains. This makes it particularly suitable for people recovering from injury or surgery, for whom the use of high training loads is unfeasible. In this article, the mechanism behind blood flow restriction training and its applicability for the treatment of lateral elbow tendinopathy is explained. An ongoing prospective, randomized, controlled trial on the treatment of lateral elbow tendinopathy is presented.

Utilization of delactosed whey permeate for the synthesis of ethyl acetate with Kluyveromyces marxianus.

Ethyl acetate is an important organic solvent and currently produced from fossil carbon resources. Microbial synthesis of this ester from sugar-rich waste could be an interesting alternative. Therefore, synthesis of ethyl acetate by Kluyveromyces marxinanus DSM 5422 from delactosed whey permeate (DWP) was studied in an aerated stirred bioreactor at 40 °C. DWP is mainly composed of residual lactose and minerals. The minerals inhibited yeast growth, as witnessed by an increased lag period, a reduced growth rate, and an extended process duration. All experiments were therefore carried out with diluted DWP. In a series of batch experiments, the pH of iron-deficient DWP medium varied between 4.8 and 5.9. The pH of the cultivation medium significantly influenced cell growth and product syntheses, with the highest ethyl acetate yield of 0.347 g g-1 and lowest by-product formation achieved at pH 5.1. It is likely that this effect is due to pH-dependent iron chelation, which affects the iron bioavailability and the intracellular iron content, thus affecting growth and metabolite synthesis. The viability of yeast cells was always high despite the harsh conditions in DWP medium, which enabled extended usage of the biomass in repeated-batch and fed-batch cultivations. These two culture techniques increased the volume of DWP processed per time by 32 and 84% for the repeated-batch and the fed-batch cultivation, respectively, without a drop of the ester yield. KEY POINTS: • Delactosed whey permeate was converted to ethyl acetate with a high rate and yield. • The formation of ethyl acetate in DWP medium at iron limitation is pH-dependent. • Highly active yeasts from batch processes enabled extension as fed and repeated batch.

The course of knee extensor strength after total knee arthroplasty: a systematic review with meta-analysis and -regression.

PURPOSE: Muscular strength loss and atrophy are postoperative complications. This systematic review with meta-analysis investigated the course of on knee extensor mass and strength from pre-surgery over total knee arthroplasty to rehabilitation and recovery. METHODS: A systematic literature search was conducted in PubMed (Medline), Cochrane Library (CINAHL, Embase) and Web of Science (until 29th of June 2022). Main inclusion criteria were ≥ 1 preoperative and ≥ 1 measurement ≥ 3-months post-operation and ≥ 1 objective assessment of quadriceps strength, muscle mass or neuromuscular activity, measured at both legs. Studies were excluded if they met the following criteria: further impairment of treated extremity or of the contralateral extremity; further muscle affecting disease, or muscle- or rehabilitation-specific intervention. The Robins-I tool for non-randomized studies, and the Cochrane Rob 2 tool for randomized controlled studies were used for risk of bias rating. Pre-surgery, 3 months, 6 months and 1 year after surgery data were pooled using random effects meta-analyses (standardized mean differences, SMD, Hedge's g) in contrast to the pre-injury values. RESULTS: 1417 studies were screened, 21 studies on 647 participants were included. Thereof, 13 were non-randomized controlled trails (moderate overall risk of bias in most studies) and 7 were randomized controlled trials (high risk of bias in at least one domain in most studies). Three (k = 12 studies; SMD = - 0.21 [95% confidence interval = - 0.36 to - 0.05], I2 = 4.75%) and six (k = 9; SMD = - 0.10 [- 0.28 to - 0.08]; I2 = 0%) months after total knee arthroplasty, a deterioration in the strength of the operated leg compared with the strength of the non-operated leg was observed. One year after surgery, the operated leg was stronger in all studies compared to the preoperative values. However, this increase in strength was not significant compared to the non-operated leg (k = 6, SMD = 0.18 [- 0.18 to 0.54], I2 = 77.56%). CONCLUSION: We found moderate certainty evidence that deficits in muscle strength of the knee extensors persist and progress until 3 months post-total knee arthroplasty in patients with end-stage knee osteoarthritis. Very low certainty evidence exists that preoperatively existing imbalance of muscle strength and mass in favor of the leg not undergoing surgery is not recovered within 1 year after surgery.

Effects of low-load blood flow restriction on the venous system in comparison to traditional low-load and high-load exercises.

Purpose: Blood-Flow-Restriction (BFR) training provides the ability to achieve hypertrophy effects even though only light mechanical loads are applied. However, its impact on venous pressures and function are still unknown. Therefore, the present study investigates the influence of BFR-training on intravascular venous pressure and venous function in comparison to control exercises with low or high mechanical loads. Methods: In a randomized cross-over design, ten healthy men (27.6 ± 6.4 years) underwent three trials of unilateral knee-extensor exercise with three different training protocols, low-load- (LL-RT, 30% of the individual 1-repetition-maximum, 1RM), low-load BFR- (LL-BFR-RT, 30% 1RM, 50% limb occlusion pressure, LOP) and high-load resistance exercise (HL-RT, 75% 1RM). Exercise protocols contain about four sets of knee extension exercise (Range-of-Motion: 0-0-95°), separated by 60 s of rest. Each set was performed until volitional muscle failure. For analysis of changes in intravascular venous pressures and venous function, a venous catheter was placed at the exercising leg before each trial. Whereas venous pressures were recorded throughout the exercise trials, phlebodynamometric investigations were performed before and after each trial. Furthermore, subjective pain perception during and after exercise was accessed by visual analogue scale. One-way ANOVA was used to assess mean differences between training protocols, while two-way repeated-measures ANOVA (rANOVA; time x condition) was performed to compare changes in measures over time among conditions. Data were given as means ± standard deviation (SD). Results: In comparison to the exercise trials without venous occlusion, total workload was significantly lower in the LL-BFR-RT (LL-RT: 1745 ± 604 kg vs LL-BFR-RT: 1274 ± 237 kg vs HL-RT: 1847 ± 367 kg, p = 0.004) without indicating statistical differences in venous pressures during the exercise sets (interaction: p = 0.140) or pain perception (interaction: p = 0.574). Similarly, phlebodynamometric assessment of venous function (e.g. refill-time of the venous system pre-vs. post exercise trials-LL-RT: 29.7 ± 11.0 s vs 25.5 ± 9.6 s, LL-BFR-RT: 26.6 ± 13.0 s vs 27.3 ± 13.8 s, HL-RT: 25.9 ± 10.9 s vs 23.1 ± 8.2 s) revealed no time (p = 0.156), condition effect (p = 0.802) or their interactions (p = 0.382). Conclusion: The present study is the first one describing the acute effects of LL-BFR-RT to muscle failure on venous pressures and function in comparison to a LL- and HL-RT in the lower limbs. In contrast to the existing literature, LL-BFR-RT does not elevate the venous pressures during exercise higher than a comparative exercise without BFR and does not show any adverse effects on venous function after the exercise.

Assessing single-locus CRISPR/Cas9-based gene drive variants in the mosquito Aedes aegypti via single-generation crosses and modeling.

The yellow fever mosquito Aedes aegypti is a major vector of arthropod-borne viruses, including dengue, chikungunya, and Zika viruses. A novel approach to mitigate arboviral infections is to generate mosquitoes refractory to infection by overexpressing antiviral effector molecules. Such an approach requires a mechanism to spread these antiviral effectors through a population, for example, by using CRISPR/Cas9-based gene drive systems. Critical to the design of a single-locus autonomous gene drive is that the selected genomic locus is amenable to both gene drive and appropriate expression of the antiviral effector. In our study, we used reverse engineering to target 2 intergenic genomic loci, which had previously shown to be highly permissive for antiviral effector gene expression, and we further investigated the use of 3 promoters (nanos, ß2-tubulin, or zpg) for Cas9 expression. We then quantified the accrual of insertions or deletions (indels) after single-generation crossings, measured maternal effects, and assessed fitness costs associated with various transgenic lines to model the rate of gene drive fixation. Overall, MGDrivE modeling suggested that when an autonomous gene drive is placed into an intergenic locus, the gene drive system will eventually be blocked by the accrual of gene drive blocking resistance alleles and ultimately be lost in the population. Moreover, while genomic locus and promoter selection were critically important for the initial establishment of the autonomous gene drive, it was the fitness of the gene drive line that most strongly influenced the persistence of the gene drive in the simulated population. As such, we propose that when autonomous CRISPR/Cas9-based gene drive systems are anchored in an intergenic locus, they temporarily result in a strong population replacement effect, but as gene drive-blocking indels accrue, the gene drive becomes exhausted due to the fixation of CRISPR resistance alleles.

Microbiological Trends and Antibiotic Susceptibility Patterns in Patients with Periprosthetic Joint Infection of the Hip or Knee over 6 Years.

We sought to analyze trends of the causative pathogens and their antibiotic susceptibility patterns in patients with periprosthetic joint infections (PJI) of the hip and knee to get better insights and improve treatment. Retrospective evaluation of all consecutive patients with microbiological detection of a causative pathogen at a tertiary endoprothetic referral center between January 2016 and December 2021 in Germany was performed. Overall, 612 different microorganisms could be detected in 493 patients (hip: n = 293; knee: n = 200). Evaluation did not show a change in the relative abundance of pathogens detected, with coagulase-negative staphylococci (n = 275; 44.9%) found frequently, followed by S. aureus (n = 86; 14.1%), Enterococcus species (n = 57; 9.3%), Streptococcus species (n = 48; 7.8%), and Gram-negative bacteria (n = 80; 13.1%). Evaluation of the antibiotic susceptibilities showed increasing rates of oxacillin-resistant coagulase-negative staphylococci (60.4%; 46.8−76.7%) and piperacillin-tazobactam-resistant Gram-negative bacteria (26.5%; 0−57.1%), although statistically not significant. Resistance of Gram-positive bacteria to vancomycin (<1%) and Gram-negative microorganisms to meropenem (1.25%) remained an exception. In summary, coagulase-negative staphylococci, as the most frequent pathogen, displayed a continuously high rate of oxacillin resistance. For the highest antimicrobial coverage in the case of an empiric therapy/unknown pathogen, vancomycin might be chosen. Level of evidence: IV.

Identification of the extracellular metallo-endopeptidases ADAM and ADAMTS in the yellow fever mosquito Aedes aegypti.

The mosquito Aedes aegypti is a major vector for dengue, Zika, yellow fever, and chikungunya (CHIKV) viruses, which cause significant morbidity and mortality among human populations in the tropical regions of the world. Following ingestion of a viremic bloodmeal from a vertebrate host, an arbovirus needs to productively infect the midgut epithelium of the mosquito. De novo synthesized virions then exit the midgut by traversing the surrounding basal lamina (BL) in order to disseminate to secondary tissues and infect those. Once the salivary glands are infected, the virus is transmitted to a vertebrate host along with saliva released during probing of the mosquito. Midgut tissue distention due to bloodmeal ingestion leads to remodeling of the midgut structure and facilitates virus dissemination from the organ. Previously, we described the matrix-metalloproteinases (MMP) of Ae. aegypti as zinc ion dependent endopeptidases (Metzincins) and showed MMP activity during midgut BL rearrangement as a consequence of bloodmeal ingestion and subsequent digestion thereby affecting arbovirus dissemination from the midgut. Here we investigate the ADAM/ADAMTS of Ae. aegypti, which form another major group of multi-domain proteinases within the Metzincin superfamily and are active during extra-cellular matrix (ECM) remodeling. Seven different ADAM and five ADAMTS were identified in Ae. aegypti. The functional protein domain structures of the identified mosquito ADAM resembled those of human ADAM10, ADAM12, and ADAM17, while two of the five mosquito ADAMTS had human orthologs. Expression profiling of Ae. aegypti ADAM/ADAMTS in immature forms, whole body-females, midguts, and ovarian tissues showed transcriptional activity of the proteinases during metamorphosis, bloodmeal ingestion/digestion, and female reproduction. Custom-made antibodies to ADAM10a and ADAM12c showed that both were strongly expressed in midgut and ovarian tissues. Furthermore, transient silencing of ADAM12c significantly reduced the carcass infection rate with CHIKV at 24 h post-infection, while silencing of ADAM12a significantly increased viral titers in secondary tissues at the same time point. Our results indicate a functional specificity for several ADAM/ADAMTS in those selected mosquito tissues.

Microbiological Profiles of Patients with Periprosthetic Joint Infection of the Hip or Knee.

Periprosthetic joint infections (PJI) are one of the most devastating consequences after total joint arthroplasty. We sought to analyze the causative pathogens of patients with PJI to get better insights and improve treatment. We performed a retrospective study of all patients with PJI of the hip and knee with microbiological detection of a causative pathogen at a tertiary endoprothetic referral center between January 2016 and March 2021. A total of 432 cases with PJI (hip: n = 250; knee: n = 182) were included. The most common causative pathogen were coagulase-negative staphylococci (n = 240; 44.2%), of which Staphylococcus epidermidis (n = 144; 26.7%) was the most frequently detected, followed by S. aureus (n = 77; 14.3%) and enterococci (n = 49; 9%). Gram-negative pathogens and fungi could be detected in 21% (n = 136) and 2.4% (n = 13) of all cases. Overall, 60% of all coagulase-negative staphylococci were oxacillin-resistant, while none of these displayed to be vancomycin-resistant. In summary, the majority of pathogens in cases of PJI could be identified as coagulase-negative staphylococci. For empirical therapy vancomycin might provide the highest antimicrobial coverage in case of an unknown pathogen.

Impact of a Six-Week Prehabilitation With Blood-Flow Restriction Training on Pre- and Postoperative Skeletal Muscle Mass and Strength in Patients Receiving Primary Total Knee Arthroplasty.

Introduction: Total Knee Arthroplasty (TKA) is one of the most successful interventions in gonarthrosis, however the operation is leading to muscle atrophy and long-term muscular deficits. To enhance rehabilitation after TKA, exercise programs try to improve muscle function preoperatively, called prehabilitation. Blood-Flow-Restriction Exercises (BFRE) is a training method which is characterized by using tourniquets to reduce arterial and occlude venous blood flow simultaneously during the exercise to increase metabolic stress. The present study aimed to evaluate the effects of a 6-week prehabilitation with BFR on pre- and postoperative muscle mass, strength, and quality of life (QoL). Methods: 30 patients with end-stage gonarthrosis participated in this study. Patients were randomized into one of three groups: 1) Control-Group (CON): Standard clinical approach without prehabilitation. 2) Active-Control-Group (AC): Participation in a prehabilitation with sham-BFR. 3) BFR-Group (BFR): Participation in a prehabilitation with BFR. The prehabilitation protocol consist of a cycling-ergometer-based training performed twice per week over 6 weeks. During exercise, BFR was applied periodically three times per leg with a pressure of 40% of the individual-limb-occlusion-pressure. Measurement time points were six- (baseline), 3-weeks and 5-days before the surgery (Pre-OP), as well as three- and 6-months postoperatively. Outcome measures were muscular strength of the thigh muscles, thigh circumference as well as QoL and functional activity, examined by 6-min walking- and chair rising test. Results: Both training groups indicated significantly improved leg muscle strength following the prehabilitation period with a superior effect for the BFR-group (BFR: ∼170% vs. AC: ∼91%, p < 0.05). No significant changes in leg strength occurred in the CON (∼3%, p = 0.100). Further, patients in BFR-group indicated significantly improved skeletal muscle mass assessed by femoral circumference following prehabilitation period (∼7%, p < 0.05), while no significant changes occurred in the CON (-1.14%, p = 0.131) and AC-group (∼3%, p = 0.078). At 3-months Post-OP, the CON and BFR-group revealed a significant decrease in femoral circumference compared to the Pre-OP (CON: ∼3%, BFR: ∼4%; p < 0.05), but BFR-group remained above the baseline level (∼3%, p < 0.05). No significant change in femoral circumference was found for AC-group (∼2%, p = 0.078). In addition, the prehabilitation with BFR provided notably improved Knee Injury and Osteoarthritis Outcome Scores (KOOS) especially in pain perception with significant higher effect compared to other groups (CON: -2%, AC: 13%, BFR: 41%; p < 0.05). In long-term rehabilitation after 6-months, all groups showed significantly improved KOOS scores in all dimensions (CON: ∼110%, AC: ∼132%, BFR: ∼225%; p < 0.01), and functional examinations (CON: ∼26%, AC: ∼16%, BFR: ∼53%; p < 0.01). Conclusion: The present findings show that BFR-prehabilitation induce significant improvements in muscle function and QoL before TKA surgery. In addition, the supporting effect of prehabilitation on postoperative regeneration and QoL should be highlighted, illustrating prolonged beneficial effects of BFR on muscular and functional performance in a "better in, better out"-manner.

Most subjectively affected joints in patients with haemophilia - what has changed after 20 years in Germany?

BACKGROUND: In patients with haemophilia (PwH), most frequently affected joints are the ankle, knee and elbow. Due to improved factor therapy in the last decades, these previous findings have to be verified in Germany. AIM: The aim of this study is to detect the most affected joint, evaluate the significance of the source of pain and determine the point prevalence of back pain in Germany today. PATIENTS AND METHODS: In a retrospective study, data of n = 300 patients with severe moderate and mild haemophilia were evaluated regarding the most affected joint, the most common source of pain, and the point prevalence of back pain. An anamnesis questionnaire and the German Pain Questionnaire were used for this assessment. RESULTS: The most affected joint in German PwH is still the ankle (41%), followed by the knee (27%) and the elbow (11%). The most common source of pain is also the ankle joint (32%). Back pain was also identified as one of the most common sources of pain, which is comparable to the elbow (elbow:15%; back:13%). The point prevalence in PwH for back pain was significantly higher compared to the general German population (P = .031). CONCLUSION: Our data showed that the ankle is still the most affected joint and the most common source of pain in Germany. These results also showed the relevance of back pain as a pain source. The evaluations also demonstrated the high point prevalence of back pain in PwH. Future therapies should also focus on the spine because joint changes affect posture.

Cellular diversity and gene expression profiles in the male and female brain of Aedes aegypti.

BACKGROUND: Aedes aegypti is a medically-important mosquito vector that transmits arboviruses including yellow fever, dengue, chikungunya, and Zika viruses to humans. The mosquito exhibits typical sexually dimorphic behaviors such as courtship, mating, host seeking, bloodfeeding, and oviposition. All these behaviors are mainly regulated by the brain; however, little is known about the function and neuron composition of the mosquito brain. In this study, we generated an initial atlas of the adult male and female brain of Ae. aegypti using 10xGenomics based single-nucleus RNA sequencing. RESULTS: We identified 35 brain cell clusters in male and female brains, and 15 of those clusters were assigned to known cell types. Identified cell types include glia (astrocytes), Kenyon cells, (ventral) projection neurons, monoaminergic neurons, medulla neurons, and proximal medulla neurons. In addition, the cell type compositions of male and female brains were compared to each other showing that they were quantitatively distinct, as 17 out of 35 cell clusters varied significantly in their cell type proportions. Overall, the transcriptomes from each cell cluster looked very similar between the male and female brain as only up to 25 genes were differentially expressed in these clusters. The sex determination factor Nix was highly expressed in neurons and glia of the male brain, whereas doublesex (dsx) was expressed in all neuron and glia cell clusters of the male and female brain. CONCLUSIONS: An initial cell atlas of the brain of the mosquito Ae. aegypti has been generated showing that the cellular compositions of the male and female brains of this hematophagous insect differ significantly from each other. Although some of the rare brain cell types have not been detected in our single biological replicate, this study provides an important basis for the further development of a complete brain cell atlas as well as a better understanding of the neurobiology of the brains of male and female mosquitoes and their sexually dimorphic behaviors.

Starvation at the larval stage increases the vector competence of Aedes aegypti females for Zika virus.

Aedes aegypti is the primary vector of Zika virus (ZIKV), a flavivirus which typically presents itself as febrile-like symptoms in humans but can also cause neurological and pregnancy complications. The transmission cycle of mosquito-borne arboviruses such as ZIKV requires that various key tissues in the female mosquito get productively infected with the virus before the mosquito can transmit the virus to another vertebrate host. Following ingestion of a viremic blood-meal from a vertebrate, ZIKV initially infects the midgut epithelium before exiting the midgut after blood-meal digestion to disseminate to secondary tissues including the salivary glands. Here we investigated whether smaller Ae. aegypti females resulting from food deprivation as larvae exhibited an altered vector competence for blood-meal acquired ZIKV relative to larger mosquitoes. Midguts from small 'Starve' and large 'Control' Ae. aegypti were dissected to visualize by transmission electron microscopy (TEM) the midgut basal lamina (BL) as physical evidence for the midgut escape barrier showing Starve mosquitoes with a significantly thinner midgut BL than Control mosquitoes at two timepoints. ZIKV replication was inhibited in Starve mosquitoes following intrathoracic injection of virus, however, Starve mosquitoes exhibited a significantly higher midgut escape and population dissemination rate at 9 days post-infection (dpi) via blood-meal, with more virus present in saliva and head tissue than Control by 10 dpi and 14 dpi, respectively. These results indicate that Ae. aegypti developing under stressful conditions potentially exhibit higher midgut infection and dissemination rates for ZIKV as adults, Thus, variation in food intake as larvae is potentially a source for variable vector competence levels of the emerged adults for the virus.