Investigation of the Microbiome of Industrial PDO Sfela Cheese and Its Artisanal Variants Using 16S rDNA Amplicon Sequencing and Shotgun Metagenomics.

Sfela is a white brined Greek cheese of protected designation of origin (PDO) produced in the Peloponnese region from ovine, caprine milk, or a mixture of the two. Despite the PDO status of Sfela, very few studies have addressed its properties, including its microbiology. For this reason, we decided to investigate the microbiome of two PDO industrial Sfela cheese samples along with two non-PDO variants, namely Sfela touloumotiri and Xerosfeli. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS), 16S rDNA amplicon sequencing and shotgun metagenomics analysis were used to identify the microbiome of these traditional cheeses. Cultured-based analysis showed that the most frequent species that could be isolated from Sfela cheese were Enterococcus faecium, Lactiplantibacillus plantarum, Levilactobacillus brevis, Pediococcus pentosaceus and Streptococcus thermophilus. Shotgun analysis suggested that in industrial Sfela 1, Str. thermophilus dominated, while industrial Sfela 2 contained high levels of Lactococcus lactis. The two artisanal samples, Sfela touloumotiri and Xerosfeli, were dominated by Tetragenococcus halophilus and Str. thermophilus, respectively. Debaryomyces hansenii was the only yeast species with abundance > 1% present exclusively in the Sfela touloumotiri sample. Identifying additional yeast species in the shotgun data was challenging, possibly due to their low abundance. Sfela cheese appears to contain a rather complex microbial ecosystem and thus needs to be further studied and understood. This might be crucial for improving and standardizing both its production and safety measures.

Early microvascular coronary endothelial dysfunction precedes pembrolizumab-induced cardiotoxicity. Preventive role of high dose of atorvastatin.

Immune checkpoint inhibitors (ICIs) exhibit remarkable antitumor activity and immune-related cardiotoxicity of unknown pathomechanism. The aim of the study was to investigate the ICI class-dependent cardiotoxicity in vitro and pembrolizumab's (Pem's) cardiotoxicity in vivo, seeking for translational prevention means. Cytotoxicity was investigated in primary cardiomyocytes and splenocytes, incubated with ipilimumab, Pem and avelumab. Pem's cross-reactivity was assessed by circular dichroism (CD) on biotechnologically produced human and murine PD-1 and in silico. C57BL6/J male mice received IgG4 or Pem for 2 and 5 weeks. Echocardiography, histology, and molecular analyses were performed. Coronary blood flow velocity mapping and cardiac magnetic resonance imaging were conducted at 2 weeks. Human EA.hy926 endothelial cells were incubated with Pem-conditioned media from human mononuclear cells, in presence and absence of statins and viability and molecular signaling were assessed. Atorvastatin (20 mg/kg, daily) was administered in vivo, as prophylaxis. Only Pem exerted immune-related cytotoxicity in vitro. Pem's cross-reactivity with the murine PD-1 was confirmed by CD and docking. In vivo, Pem initiated coronary endothelial and diastolic dysfunction at 2 weeks and systolic dysfunction at 5 weeks. At 2 weeks, Pem induced ICAM-1 and iNOS expression and intracardiac leukocyte infiltration. At 5 weeks, Pem exacerbated endothelial activation and triggered cardiac inflammation. Pem led to immune-related cytotoxicity in EA.hy926 cells, which was prevented by atorvastatin. Atorvastatin mitigated functional deficits, by inhibiting endothelial dysfunction in vivo. We established for the first time an in vivo model of Pem-induced cardiotoxicity. Coronary endothelial dysfunction precedes Pem-induced cardiotoxicity, whereas atorvastatin emerges as a novel prophylactic therapy.

Higher Infection Rate after ACL Reconstruction with Hamstrings Tendon Autografts Compared with Bone Patellar Bone Tendon Autografts: A Review.

Infection after anterior cruciate ligament reconstruction is a rare but devastating complication resulting in a deleterious impact on knee function as well as an increased related cost for treatment and rehabilitation for the patients. There are conflicting reports regarding the rate of infection between bone patellar tendon bone (BPTB) and hamstrings tendon (HT) autografts for anterior cruciate ligament reconstruction. Therefore, we performed this review to summarize all the available data regarding the risk of infection after ACL reconstruction, to provide insight on the infection risk between BPTB and HT autografts, and to discuss current recommendations for the diagnosis and treatment of these infections. The incidence and risk of infection after ACL reconstruction with HT graft is higher compared with BPTB grafts. The most commonly subacute and late infections, quadruple type, need for cannulated instruments for harvesting, size and shape and fixation of the extra-tunnel material of the HT are important risk factors for infection. Combined antibiotics administration and adequate arthroscopic lavage and debridement are the optimal treatments for ACL reconstruction infection. Early diagnosis and treatment is the most important predictor for graft retention, which accounts more commonly for BPTB grafts. The treating physicians should be aware of the higher incidence of infection after ACL reconstruction with HT graft, as well as for the need for a high clinical suspicion for early diagnosis of the infection. These will increase the possibility of eradication of the infection and retention of the graft.

Effect of laterality and fatigue in peroneal electromechanical delay.

INTRODUCTION: Extremity dominance is one of the intrinsic factors that have been identified for ankle sprains. Electromechanical delay (EMD) is an integral part of the peroneal motor response and, therefore, substantial in preventing ankle sprains. This study aimed to investigate the effect of laterality on EMD times before and after fatigue. METHODS: Fifteen healthy male volunteers participated in the study. Measurements were taken with the ankle in a neutral (0°) position, and all subjects followed an isokinetic fatigue protocol. Repeated ANOVA was used for statistical analysis, and the α level was set a priori at p ≤ 0.05. RESULTS: No significant difference was noted in EMD times between the dominant and non-dominant legs of the volunteers (p = 0.940). Fatigue caused a significant increase in EMD by 10-15 ms (p = 0.003), while the leg × fatigue interaction was not significant (p = 0.893). CONCLUSIONS: In a non-injured athlete, both ankles seem to be under the same protection of the reactive response of the peroneal muscles. Therefore, athletes should be aware that both their extremities are equally exposed to the danger of an ankle injury. Also, fatigued ankles demonstrate longer EMD times, implying that improving resistance to fatigue may add another layer of protection that has the potential to prevent ankle sprain recurrence.

Acute administration of the olive constituent, oleuropein, combined with ischemic postconditioning increases myocardial protection by modulating oxidative defense.

Oleuropein, one of the main polyphenolic constituents of olive, is cardioprotective against ischemia reperfusion injury (IRI). We aimed to assess the cardioprotection afforded by acute administration of oleuropein and to evaluate the underlying mechanism. Importantly, since antioxidant therapies have yielded inconclusive results in attenuating IRI-induced damage on top of conditioning strategies, we investigated whether oleuropein could enhance or imbed the cardioprotective manifestation of ischemic postconditioning (PostC). Oleuropein, given during ischemia as a single intravenous bolus dose reduced the infarct size compared to the control group both in rabbits and mice subjected to myocardial IRI. None of the inhibitors of the cardioprotective pathways, l-NAME, wortmannin and AG490, influence its infarct size limiting effects. Combined oleuropein and PostC cause further limitation of infarct size in comparison with PostC alone in both animal models. Oleuropein did not inhibit the calcium induced mitochondrial permeability transition pore opening in isolated mitochondria and did not increase cGMP production. To provide further insights to the different cardioprotective mechanism of oleuropein, we sought to characterize its anti-inflammatory potential in vivo. Oleuropein, PostC and their combination reduce inflammatory monocytes infiltration into the heart and the circulating monocyte cell population. Oleuropein's mechanism of action involves a direct protective effect on cardiomyocytes since it significantly increased their viability following simulated IRI as compared to non-treated cells. Οleuropein confers additive cardioprotection on top of PostC, via increasing the expression of the transcription factor Nrf-2 and its downstream targets in vivo. In conclusion, acute oleuropein administration during ischemia in combination with PostC provides robust and synergistic cardioprotection in experimental models of IRI by inducing antioxidant defense genes through Nrf-2 axis and independently of the classic cardioprotective signaling pathways (RISK, cGMP/PKG, SAFE).

A computer tablet software can quantify the deviation of scapula medial border from the thoracic wall during clinical assessment of scapula dyskinesis.

PURPOSE: Aim of this study is to establish an objective and easily applicable method that will allow clinicians to quantitatively assess scapular dyskinesis during clinical examination using a computer tablet software. Hypothesis is that dyskinetic scapulae present greater motion-deviation from the thoracic wall-compared to the non-dyskinetic ones and that the software will be able to record those differences. METHODS: Twenty-five patients and 19 healthy individuals were clinically evaluated for the presence of dyskinesis or not. According to the clinical diagnosis, the observations were divided into three groups; A. Dyskinetic scapulae with symptoms (n = 25), B. Contralateral non-dyskinetic scapulae without symptoms (n = 25), C. Healthy control scapulae (n = 38). Then, all individuals were tested using a tablet with the PIVOT™ image-based analysis software (PIVOT, Impellia, Pittsburgh, PA, USA). The motion produced by the scapula medial border and inferior angle deviation from the thoracic wall was recorded. RESULTS: The deviation of the medial border and inferior angle of the scapula from the thoracic wall was 24.6 ± 7.3 mm in Group A, 14.7 ± 4.9 mm in Group B, and 12.4 ± 5.2 mm in Group C. The motion recorded in the dyskinetic scapulae group was significantly greater than both the contralateral non-dyskinetic scapulae group (p < 0.01) and the healthy control scapulae group (p < 0.01). CONCLUSION: The PIVOT™ software was efficient to detect significant differences in the motion between dyskinetic and non-dyskinetic scapulae. This system can support the clinical diagnosis of dyskinesis with a numeric value, which not only contributes to scapula dyskinesis grading but also to the evaluation of the progress and efficacy of the applied treatment, thus providing a feedback to the clinician and the patient. LEVEL OF EVIDENCE: IV, laboratory study.

Screening of Heteroaromatic Scaffolds against Cystathionine Beta-Synthase Enables Identification of Substituted Pyrazolo[3,4-c]Pyridines as Potent and Selective Orthosteric Inhibitors.

Cystathionine ß-synthase (CBS) is a key enzyme in the production of the signaling molecule hydrogen sulfide, deregulation of which is known to contribute to a range of serious pathological states. Involvement of hydrogen sulfide in pathways of paramount importance for cellular homeostasis renders CBS a promising drug target. An in-house focused library of heteroaromatic compounds was screened for CBS modulators by the methylene blue assay and a pyrazolopyridine derivative with a promising CBS inhibitory potential was discovered. The compound activity was readily comparable to the most potent CBS inhibitor currently known, aminoacetic acid, while a promising specificity over the related cystathionine γ-lyase was identified. To rule out any possibility that the inhibitor may bind the enzyme regulatory domain due to its high structural similarity with cofactor s-adenosylmethionine, differential scanning fluorimetry was employed. A sub-scaffold search guided follow-up screening of related compounds, providing preliminary structure-activity relationships with respect to requisites for efficient CBS inhibition by this group of heterocycles. Subsequently, a hypothesis regarding the exact binding mode of the inhibitor was devised on the basis of the available structure-activity relationships (SAR) and a deep neural networks analysis and further supported by induced-fit docking calculations.

Does Manual Drilling Improve the Healing of Bone-Hamstring Tendon Grafts in Anterior Cruciate Ligament Reconstruction? A Histological and Biomechanical Study in a Rabbit Model.

BACKGROUND: Heat necrosis due to motorized drilling during anterior cruciate ligament (ACL) reconstruction could be a factor in delayed healing at the bone-tendon graft interface. HYPOTHESIS: The process of osteointegration could be enhanced using manual drilling. It reduces the invasiveness of mechanical-thermal stress normally caused by the traditional motorized drill bit. STUDY DESIGN: Controlled laboratory study. METHODS: ACL reconstruction using semitendinosus tendon autografts was performed in 28 skeletally mature female New Zealand white rabbits, which were randomly divided into 3 groups. In group A (n = 12), the tunnels were drilled using a motorized device; in group B (n = 12), the tunnels were drilled using a manual drill bit; and group C (n = 4) served as a control with sham surgical procedures. The healing process in the tunnels was assessed histologically at 2, 4, 8, and 12 weeks and graded according to the Tendon-Bone Tunnel Healing (TBTH) scoring system. In addition, another 25 rabbits were used for biomechanical testing. The structural properties of the femur-ACL graft-tibia complex, from animals sacrificed at 8 weeks postoperatively, were determined using uniaxial tests. Stiffness (N/mm) and ultimate load to failure (N) were determined from the resulting load-elongation curves. RESULTS: The time course investigation showed that manual drilling (group B) had a higher TBTH score and improved mechanical behavior, reflecting better organized collagen fiber continuity at the bone-fibrous tissue interface, better integration between the graft and bone, and early mineralized chondrocyte-like tissue formation at all the time points analyzed with a maximum difference at 4 weeks (TBTH score: 5.4 [group A] vs 12.3 [group B]; P < .001). Stiffness (23.1 ± 8.2 vs 17.8 ± 6.3 N/mm, respectively) and ultimate load to failure (91.8 ± 60.4 vs 55.0 ± 18.0 N, respectively) were significantly enhanced in the specimens treated with manual drilling compared with motorized drilling (P < .05 for both). CONCLUSION: The use of manual drilling during ACL reconstruction resulted in better tendon-to-bone healing during the crucial early weeks. Manual drilling was able to improve the biological and mechanical properties of bone-hamstring tendon graft healing and was able to restore postoperative graft function more quickly. Tunnel drilling results in bone loss and deficient tendon-bone healing, and heat necrosis after tunnel enlargement may cause mechanical stress, contributing to a delay in healing. Manual drilling preserved the bone stock inside the tunnel, reduced heat necrosis, and offered a better microenvironment for faster healing at the interface. CLINICAL RELEVANCE: Based on study results, manual drilling could be used successfully in human ACL reconstruction, but further clinical studies are needed. A clinical alternative, called the original "all-inside" technique, has been developed for ACL reconstruction. In this technique, the femoral and tibial tunnels are manually drilled only halfway through the bone for graft fixation, reducing bone loss. Data from this study suggest that hamstring tendon-to-bone healing can be improved using a manual drilling technique to form femoral and tibial tunnels.

Complete Genome Sequence of the Deep-Sea Bacterium Moritella marina MP-1 (ATCC 15381).

Here, the complete assembly of the Moritella marina MP-1 (ATCC 15381) genome, combining Illumina and long Nanopore reads, is presented. The gapless assembly consists of a 4.7-Mb circular chromosome and a 26-kb plasmid, with a G+C content of 40.7%, and will assist in further studies of the molecular pathways in this biotechnologically significant organism.

Consensus protein engineering on the thermostable histone-like bacterial protein HUs significantly improves stability and DNA binding affinity.

Consensus-based protein engineering strategy has been applied to various proteins and it can lead to the design of proteins with enhanced biological performance. Histone-like HUs comprise a protein family with sequence variety within a highly conserved 3D-fold. HU function includes compacting and regulating bacterial DNA in a wide range of biological conditions in bacteria. To explore the possible impact of consensus-based design in the thermodynamic stability of HU proteins, the approach was applied using a dataset of sequences derived from a group of 40 mesostable, thermostable, and hyperthermostable HUs. The consensus-derived HU protein was named HUBest, since it is expected to perform best. The synthetic HU gene was overexpressed in E. coli and the recombinant protein was purified. Subsequently, HUBest was characterized concerning its correct folding and thermodynamic stability, as well as its ability to interact with plasmid DNA. A substantial increase in HUBest stability at high temperatures is observed. HUBest has significantly improved biological performance at ambience temperature, presenting very low Kd values for binding plasmid DNA as indicated from the Gibbs energy profile of HUBest. This Kd may be associated to conformational changes leading to decreased thermodynamic stability and, therefore, higher flexibility at ambient temperature.

Treatment of SLAP Lesions.

BACKGROUND: The surgical treatment of a Superior Labrum Anterior and Posterior (SLAP) lesion becomes more and more frequent as the surgical techniques, the implants and the postoperative rehabilitation of the patient are improved and provide in most cases an excellent outcome. OBJECTIVE: However, a standard therapy of SLAP lesions in the shoulder surgery has not been established yet. An algorithm on how to treat SLAP lesions according to their type and data on the factors that influence the surgical outcome is essential for the everyday clinical practice. METHOD: In this article, a retrospective evaluation of patients with SLAP lesion, treated surgically in our orthopaedic clinic was conducted. RESULTS: According to the clinical outcome and our experience with the surgical therapy of SLAP lesions we demonstrate an algorithm on the proper therapeutic approach. CONCLUSION: SLAP I lesions are treated with debridement. Most controversies concern patients with SLAP II lesions, whose therapy is either fixation of the superior labrum or tenotomy/tenodesis of the long head of the biceps tendon. For patients with SLAP III or IV lesions the most commonly accepted approach is tenotomy or tenodesis of the long head of biceps tendon.

Restoring tibiofemoral alignment during ACL reconstruction results in better knee biomechanics.

PURPOSE: Anterior cruciate ligament (ACL) reconstruction (ACLR) aims to restore normal knee joint function, stability and biomechanics and in the long term avoid joint degeneration. The purpose of this study is to present the anatomic single bundle (SB) ACLR that emphasizes intraoperative correction of tibiofemoral subluxation that occurs after ACL injury. It was hypothesized that this technique leads to optimal outcomes and better restoration of pathological tibiofemoral joint movement that results from ACL deficiency (ACLD). METHODS: Thirteen men with unilateral ACLD were prospectively evaluated before and at a mean follow-up of 14.9 (SD = 1.8) months after anatomic SB ACLR with bone patellar tendon bone autograft. The anatomic ACLR replicated the native ACL attachment site anatomy and graft orientation. Emphasis was placed on intraoperative correction of tibiofemoral subluxation by reducing anterior tibial translation (ATT) and internal tibial rotation. Function was measured with IKDC, Lysholm and the Tegner activity scale, ATT was measured with the KT-1000 arthrometer and tibial rotation (TR) kinematics were measured with 3Dmotion analysis during a high-demand pivoting task. RESULTS: The results showed significantly higher TR of the ACL-deficient knee when compared to the intact knee prior to surgery (12.2° ± 3.7° and 10.7° ± 2.6° respectively, P = 0.014). Postoperatively, the ACLR knee showed significantly lower TR as compared to the ACL-deficient knee (9.6°±3.1°, P = 0.001) but no difference as compared to the control knee (n.s.). All functional scores were significantly improved and ATT was restored within normal values (P < 0.001). CONCLUSIONS: Intraoperative correction of tibiofemoral subluxation that results after ACL injury is an important step during anatomic SB ACLR. The intraoperative correction of tibiofemoral subluxation along with the replication of native ACL anatomy results in restoration of rotational kinematics of ACLD patients to normal levels that are comparable to the control knee. These results indicate that the reestablishment of tibiofemoral alignment during ACLR may be an important step that facilitates normal knee kinematics postoperatively. LEVEL OF EVIDENCE: Level II, prospective cohort study.

Genome Instability and γH2AX.

γH2AX has emerged in the last 20 years as a central player in the DDR (DNA damage response), with specificity for DSBs (double-strand breaks). Upon the generation of DSBs, γ-phosphorylation extends along megabase-long domains in chromatin, both sides of the damage. The significance of this mechanism is of great importance; it depicts a biological amplification mechanism where one DSB induces the γ-phosphorylation of thousands of H2AX molecules along megabaselong domains of chromatin, that are adjusted to the sites of DSBs. A sequential recruitment of signal transduction factors that interact to each other and become activated to further amplify the signal that will travel to the cytoplasm take place on the γ-phosphorylated chromatin. γ-phosphorylation is an early event in the DSB damage response, induced in all phases of the cell cycle, and participates in both DSB repair pathways, the HR (homologous recombination) and NHEJ (non-homologous end joining). Today, numerous studies support the notion that γH2AX functions as a guardian of the genome by preventing misrepaired DSB that increase the mutation load of the cells and may further lead to genome instability and carcinogenesis.

Peroneal electromechanical delay and fatigue in patients with chronic ankle instability.

PURPOSE: The purpose of this study was to investigate the effect of chronic ankle instability (CAI) on electromechanical delay times (EMD) before and after fatigue. Understanding the mechanisms that contribute to CAI is essential for the development of effective rehabilitation programmes. It was hypothesized that patients with CAI will demonstrate prolonged EMD times compared to healthy subjects and that fatigue will cause greater increases in EMD times in the CAI group. METHODS: Twenty-one male volunteers participated in the study providing data on 16 ankles with CAI and 26 with no history of ankle injury. EMD was measured on an isokinetic dynamometer. Measurements were taken with the ankle in neutral (0°) and at 30° of inversion. All subjects followed an isokinetic fatigue protocol until eversion torque fell below 50 % of initial torque for three consecutive repetitions. A 2 × 2 × 2 ANOVA was used to calculate the effect of ankle status (CAI vs. healthy), fatigue, angle (0° vs. 30°) and their interactions on EMD. RESULTS: Fatigue caused a significant increase on EMD [non-fatigued: 122(29)ms vs. fatigue 155(54)ms; p < 0.001]. EMD times were shorter at 30° of inversion compared to neutral [neutral: 145(39)ms vs. 30° of inversion: 132(40)ms, p = 0.015]. An interaction effect for ankle status and angle was found (p = 0.026) with CAI ankles demonstrating longer EMD [CAI: 156(45)ms vs. healthy: 133(40)ms] in neutral but not at 30° of inversion [CAI: 133(46)ms vs. 132(33)ms]. CONCLUSIONS: Patients with CAI had longer EMD times in neutral, but not when the ankle was placed in inversion. This suggests that rehabilitation programmes may be more effective when retraining occurs with the ankle in neutral position. It is likely that low EMD times prevent ankle acceleration at the beginning of the mechanism of injury, but they are less important when the ankle has already inverted at 30°. Both CAI and healthy subjects demonstrated longer EMD after fatigue, emphasizing the importance of proper conditioning in the prevention of delayed peroneal response and subsequent ankle injury. Improving resistance to fatigue of the peroneals may prove to be an effective prevention tool of ankle sprain recurrence in patients with CAI. LEVEL OF EVIDENCE: III.

Generation of stem cell-based bioartificial anterior cruciate ligament (ACL) grafts for effective ACL rupture repair.

In the present study, we combined stem cell technology with a non-absorbable biomaterial for the reconstruction of the ruptured ACL. Towards this purpose, multipotential stromal cells derived either from subcutaneous human adipose tissue (hAT-MSCs) or from induced pluripotent stem cells (iPSCs) generated from human foreskin fibroblasts (hiPSC-MSCs) were cultured on the biomaterial for 21days in vitro to generate a 3D bioartifical ACL graft. Stem cell differentiation towards bone and ligament at the ends and central part of the biomaterial was selectively induced using either BMP-2/FGF-2 or TGF-ß/FGF-2 combinations, respectively. The bioartificial ACL graft was subsequently implanted in a swine ACL rupture model in place of the surgically removed normal ACL. Four months post-implantation, the tissue engineered ACL graft generated an ACL-like tissue exhibiting morphological and biochemical characteristics resembling those of normal ACL.

Insights into the functionality and stability of designer cellulosomes at elevated temperatures.

Enzymatic breakdown of lignocellulose is a major limiting step in second generation biorefineries. Assembly of the necessary activities into designer cellulosomes increases the productivity of this step by enhancing enzyme synergy through the proximity effect. However, most cellulosomal components are obtained from mesophilic microorganisms, limiting the applications to temperatures up to 50 °C. We hypothesized that a scaffoldin, comprising modular components of mainly mesophilic origin, can function at higher temperatures when combined with thermophilic enzymes, and the resulting designer cellulosomes could be employed in higher temperature reactions. For this purpose, we used a tetravalent scaffoldin constituted of three cohesins of mesophilic origin as well as a cohesin and cellulose-binding module derived from the thermophilic bacterium Clostridium thermocellum. The scaffoldin was combined with four thermophilic enzymes from Geobacillus and Caldicellulosiruptor species, each fused with a dockerin whose specificity matched one of the cohesins. We initially verified that the biochemical properties and thermal stability of the resulting chimeric enzymes were not affected by the presence of the mesophilic dockerins. Then we examined the stability of the individual single-enzyme-scaffoldin complexes and the full tetravalent cellulosome showing that all complexes are stable and functional for at least 6 h at 60 °C. Finally, within this time frame and conditions, the full complex appeared over 50 % more efficient in the hydrolysis of corn stover compared to the free enzymes. Overall, the results support the utilization of scaffoldin components of mesophilic origin at relatively high temperatures and provide a framework for the production of designer cellulosomes suitable for high temperature biorefinery applications.

Analysis of the complete genome sequence of the archaeon Pyrococcus chitonophagus DSM 10152 (formerly Thermococcus chitonophagus).

Here we analyze the first complete genome sequence of Pyrococcus chitonophagus. The archaeon was previously suggested to belong to the Thermococcus rather than the Pyrococcus genus. Whole genome phylogeny as well as whole proteome comparisons using all available complete genomes in Thermococcales clearly showed that the species belongs to the Pyrococcus genus. P. chitonophagus was originally isolated from a hydrothermal vent site and it has been described to effectively degrade chitin debris, and therefore is considered to play a major role in the sea water ecology and metabolic activity of microbial consortia within hot sea water ecosystems. Indeed, an obvious feature of the P. chitonophagus genome is that it carries proteins showing complementary activities for chitin degradation, i.e. endo- and exo-chitinase, diacetylchitobiose deacetylase and exo-ß-D glucosaminidase activities. This finding supports the hypothesis that compared to other Thermococcales species P. chitonophagus is adapted to chitin degradation.

Comparison of knee flexion isokinetic deficits between seated and prone positions after ACL reconstruction with hamstrings graft: Implications for rehabilitation and return to sports decisions.

OBJECTIVES: Hamstrings grafts are commonly used in ACL reconstruction, however, the effect of graft harvesting on knee flexion strength has not been longitudinally evaluated in functional positions. We hypothesized that greater deficits in knee flexion strength exist in the prone compared to the seated position and these deficits remain as rehabilitation progresses. DESIGN: Case series. METHODS: Forty-two consecutive patients who underwent ACL reconstruction with a hamstrings graft were followed prospectively for 9 months. Isokinetic knee flexion strength at a slow and a fast speed were collected at 3, 4, 6, and 9 months in two different positions: conventional (seated) and functional (0° of hip flexion). RESULTS: Peak torque knee flexion deficits were higher in the prone position compared to the seated position by an average of 6.5% at 60°/s and 9.1% at 180°/s (p<0.001). CONCLUSIONS: Measuring knee flexion strength in prone demonstrates higher deficits than in the conventional seated position. Most athletes would not be cleared to return to sports even at 9 months after surgery with this method.

Electromechanical delay of the knee flexor muscles after anterior cruciate ligament reconstruction using semitendinosus tendon.

The purpose of the study was to evaluate whether using only the semitendinosus as a tripled short graft would affect the electromechanical delay (EMD) of the knee flexors. EMD was evaluated in volunteers (N = 15) after they had undergone surgery for anterior cruciate ligament (ACL) reconstruction where the semitendinosus tendon alone was used as a graft. The results were compared with the intact leg and healthy controls (N = 15). After warming up, each subject performed four maximally explosive isometric contractions on an isokinetic dynamometer. Torques were measured by the dynamometer, while the electrical activity of the semitendinosus and biceps femoris muscles was detected using surface electromyography. EMD was found to be significantly increased (p = 0.001) in patients who had undergone ACL reconstruction compared to the controls. On the contrary, no significant differences (p = 0.235) were found for the biceps femoris muscle between the two groups. Similar results were found when the study group was compared with the intact leg group (p = 0.027 for semitendinosus and p = 0.859 for biceps femoris). Harvesting the semitendinosus tendon increases the EMD for the semitendinosus muscle but does not influence the EMD outcomes for the biceps femoris muscle.

Association of the Single-Limb Hop Test With Isokinetic, Kinematic, and Kinetic Asymmetries in Patients After Anterior Cruciate Ligament Reconstruction.

BACKGROUND: Asymmetries persist after anterior cruciate ligament reconstruction (ACLR). Physical performance tests such as the single-limb hop test have been used extensively to assess return-to-sport criteria, as they reproduce dynamic athletic maneuvers. HYPOTHESIS: The single-limb hop is associated with muscle strength and kinematic and kinetic asymmetries in ACLR patients 6 to 9 months after surgery. STUDY DESIGN: Controlled laboratory study. METHODS: Twenty-two men with ACLR (mean age, 28.8 ± 11.2 years) at 6 to 9 months (mean, 7.01 ± 0.93 months) after surgery completed isokinetic testing in 3 velocities (120, 180, and 300 deg/s) and a kinetic, kinematic, and functional evaluation of the single-limb hop test. Pearson correlation coefficients were used to assess the relationship between the Limb Symmetry Index (LSI) of the single-limb hop distance and each of the outcome variables. RESULTS: There were significant positive correlations between the LSI of the single-limb hop distance and the LSI of the peak extension torque at 120 deg/s (P = 0.044, r = 0.37) and the peak extension torque at 180 deg/s (P = 0.042, r = 0.38) as well as a negative correlation with the peak flexion torque at 180 deg/s (P = 0.043, r = -0.38). The LSI of the single-limb hop test was not correlated with any kinetic or kinematic variable (P > 0.05). CONCLUSION: The findings of the present study demonstrate that distance LSI of the single-limb hop test correlates with isokinetic extension peak torque LSI but not kinetic and kinematic asymmetry. CLINICAL RELEVANCE: The single-limb hop test can be used as an additional tool for the recognition of muscle strength asymmetries but not for kinetic or kinematic asymmetries 6 to 9 months after ACLR.