Morphological, ultrastructural, genetic characteristics and remarkably low prevalence of macroscopic Sarcocystis species isolated from sheep and goats in Kurdistan region, Iraq.

Introduction: Sarcocystis is a genus of cyst-forming parasites that infest both humans and livestock. Some parasites cause clinical and subclinical diseases in their hosts, resulting in economic losses. Methods: Esophagus, diaphragm, and skeletal muscle from slaughtered sheep and goats were examined macroscopically, microscopically, and ultrastructurally and subjected to DNA analysis. Results: We isolated macrocysts of S. gigantea and of S. caprafelis moulei from naturally infected sheep (Ovis aries) and goats (Capra hircus). The macrocyst wall thickness was 18.9 µm in sheep and 15.3 µm in goats, and consisted of an inner Periodic acid Schiff- (PAS) negative primary wall and an outer glycoconjugates containing i.e. PAS-positive secondary wall. The walls inner surface was compartmentalized and filled with bradyzoites. In S. gigantea the bradyzoites were approximently 12.3 x 2.6 µm in size, while in S. caprafelis moulei they were 13.9 x 4.4 µm. Ultrastructurally, both species have nearly identical morphology: cauliflower-like protrusions with numerous microtubules and often dendritic-like filaments, branching from the primary wall. The 18S rRNA gene in S. gigantea was 85.9% identical to that in S. medusiformis and 80.4% to the S. caprafelis moulei gene. The 28S rRNA gene in S. gigantea was 94.6% identical to that in S. medusiformis and 97.3% to the S. caprafelis moulei. Conclusion: This study is the first to (i) detail the ultrastructure of the macrocyst wall of S. caprafelis moulei, (ii) identify S. medusiformis in Iraqi sheep, and (iii) compare the prevalence of macroscopic Sarcocystis at different time periods within the same region. A positive finding was the reduction of macroscopic sarcocystosis occurrences (0.01% in sheep and 0.02% in goats) compared to our previous data from 1992 (4.1%: sheep, 33.6%: goats).

Treatment With Nimodipine or FK506 After Facial Nerve Repair Neither Improves Accuracy of Reinnervation Nor Recovery of Mimetic Function in Rats.

Purpose: Nimodipine and FK506 (Tacrolimus) are drugs that have been reported to accelerate peripheral nerve regeneration. We therefore tested these substances aiming to improve the final functional outcome of motoric reinnervation after facial nerve injury. Methods: In 18 female rats, the transected facial nerve was repaired by an artificial nerve conduit. The rats were then treated with either placebo, nimodipine, or FK506, for 56 days. Facial motoneurons were pre-operatively double-labeled by Fluoro-Gold and again 56 days post-operation by Fast-Blue to measure the cytological accuracy of reinnervation. The whisking motion of the vibrissae was analyzed to assess the quality of functional recovery. Results: On the non-operated side, 93-97% of those facial nerve motoneurons innervating the vibrissae were double-labeled. On the operated side, double-labeling only amounted to 38% (placebo), 40% (nimodipine), and 39% (FK506), indicating severe misdirection of reinnervation. Regardless of post-operative drug or placebo therapy, the whisking frequency reached 83-100% of the normal value (6.0 Hz), but whisking amplitude was reduced to 33-48% while whisking velocity reached 39-66% of the normal values. Compared to placebo, statistically neither nimodipine nor FK506 improved accuracy of reinnervation and function recovery. Conclusion: Despite previous, positive data on the speed and quantity of axonal regeneration, nimodipine and FK506 do not improve the final functional outcome of motoric reinnervation in rats.

Angiotensin II type 1 receptor localizes at the blood-bile barrier in humans and pigs.

Animal models and clinical studies suggest an influence of angiotensin II (AngII) on the pathogenesis of liver diseases via the renin-angiotensin system. AngII application increases portal blood pressure, reduces bile flow, and increases permeability of liver tight junctions. Establishing the subcellular localization of angiotensin II receptor type 1 (AT1R), the main AngII receptor, helps to understand the effects of AngII on the liver. We localized AT1R in situ in human and porcine liver and porcine gallbladder by immunohistochemistry. In order to do so, we characterized commercial anti-AT1R antibodies regarding their capability to recognize heterologous human AT1R in immunocytochemistry and on western blots, and to detect AT1R using overlap studies and AT1R-specific blocking peptides. In hepatocytes and canals of Hering, AT1R displayed a tram-track-like distribution, while in cholangiocytes AT1R appeared in a honeycomb-like pattern; i.e., in liver epithelia, AT1R showed an equivalent distribution to that in the apical junctional network, which seals bile canaliculi and bile ducts along the blood-bile barrier. In intrahepatic blood vessels, AT1R was most prominent in the tunica media. We confirmed AT1R localization in situ to the plasma membrane domain, particularly between tight and adherens junctions in both human and porcine hepatocytes, cholangiocytes, and gallbladder epithelial cells using different anti-AT1R antibodies. Localization of AT1R at the junctional complex could explain previously reported AngII effects and predestines AT1R as a transmitter of tight junction permeability.

Treadmill training of rats after sciatic nerve graft does not alter accuracy of muscle reinnervation.

Background and purpose: After peripheral nerve lesions, surgical reconstruction facilitates axonal regeneration and motor reinnervation. However, functional recovery is impaired by aberrant reinnervation. Materials and methods: We tested whether training therapy by treadmill exercise (9 × 250 m/week) before (run-idle), after (idle-run), or both before and after (run-run) sciatic nerve graft improves the accuracy of reinnervation in rats. Female Lewis rats (LEW/SsNHsd) were either trained for 12 weeks (run) or not trained (kept under control conditions, idle). The right sciatic nerves were then excised and reconstructed with 5 mm of a congenic allograft. One week later, training started in the run-run and idle-run groups for another 12 weeks. No further training was conducted in the run-idle and idle-idle groups. Reinnervation was measured using the following parameters: counting of retrogradely labeled motoneurons, walking track analysis, and compound muscle action potential (CMAP) recordings. Results: In intact rats, the common fibular (peroneal) and the soleus nerve received axons from 549 ± 83 motoneurons. In the run-idle group, 94% of these motoneurons had regenerated 13 weeks after the nerve graft. In the idle-run group, 81% of the normal number of motoneurons had regenerated into the denervated musculature and 87% in both run-run and idle-idle groups. Despite reinnervation, functional outcome was poor: walking tracks indicated no functional improvement of motion in any group. However, in the operated hindlimb of run-idle rats, the CMAP of the soleus muscle reached 11.9 mV (normal 16.3 mV), yet only 6.3-8.1 mV in the other groups. Conclusion: Treadmill training neither altered the accuracy of reinnervation nor the functional recovery, and pre-operative training (run-idle) led to a higher motor unit activation after regeneration.

Caldesmon ablation in mice causes umbilical herniation and alters contractility of fetal urinary bladder smooth muscle.

The actin-, myosin-, and calmodulin-binding protein caldesmon (CaD) is expressed in two splice isoforms: h-CaD, which is an integral part of the actomyosin domain of smooth muscle cells, and l-CaD, which is widely expressed and is involved in many cellular functions. Despite extensive research for many years, CaD's in vivo function has remained elusive. To explore the role of CaD in smooth muscle contraction in vivo, we generated a mutant allele that ablates both isoforms. Heterozygous animals were viable and had a normal life span, but homozygous mutants died perinatally, likely because of a persistent umbilical hernia. The herniation was associated with hypoplastic and dysmorphic abdominal wall muscles. We assessed mechanical parameters in isometrically mounted longitudinal strips of E18.5 urinary bladders and in ring preparations from abdominal aorta using wire myography. Ca2+ sensitivity was higher and relaxation rate was slower in Cald1-/- compared with Cald1+/+ skinned bladder strips. However, we observed no change in the content and phosphorylation of regulatory proteins of the contractile apparatus and myosin isoforms known to affect these contractile parameters. Intact fibers showed no difference in actin and myosin content, regardless of genotype, although KCl-induced force tended to be lower in homozygous and higher in heterozygous mutants than in WTs. Conversely, in skinned fibers, myosin content and maximal force were significantly lower in Cald1-/- than in WTs. In KO abdominal aortas, resting and U46619 elicited force were lower than in WTs. Our results are consistent with the notion that CaD impacts smooth muscle function dually by (1) acting as a molecular brake on contraction and (2) maintaining the structural integrity of the contractile machinery. Most importantly, CaD is essential for resolution of the physiological umbilical hernia and ventral body wall closure.

Direct, gabapentin-insensitive interaction of a soluble form of the calcium channel subunit α2δ-1 with thrombospondin-4.

The α2δ-1 subunit of voltage-gated calcium channels binds to gabapentin and pregabalin, mediating the analgesic action of these drugs against neuropathic pain. Extracellular matrix proteins from the thrombospondin (TSP) family have been identified as ligands of α2δ-1 in the CNS. This interaction was found to be crucial for excitatory synaptogenesis and neuronal sensitisation which in turn can be inhibited by gabapentin, suggesting a potential role in the pathogenesis of neuropathic pain. Here, we provide information on the biochemical properties of the direct TSP/α2δ-1 interaction using an ELISA-style ligand binding assay. Our data reveal that full-length pentameric TSP-4, but neither TSP-5/COMP of the pentamer-forming subgroup B nor TSP-2 of the trimer-forming subgroup A directly interact with a soluble variant of α2δ-1 (α2δ-1S). Interestingly, this interaction is not inhibited by gabapentin on a molecular level and is not detectable on the surface of HEK293-EBNA cells over-expressing α2δ-1 protein. These results provide biochemical evidence that supports a specific role of TSP-4 among the TSPs in mediating the binding to neuronal α2δ-1 and suggest that gabapentin does not directly target TSP/α2δ-1 interaction to alleviate neuropathic pain.

Tumor-Derived Extracellular Vesicles Inhibit Natural Killer Cell Function in Pancreatic Cancer.

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal malignancies. Tumor-derived extracellular vesicles (EVs) induce pre-metastatic niche formation to promote metastasis. We isolated EVs from a highly-metastatic pancreatic cancer cell line and patient-derived primary cancer cells by ultracentrifugation. The protein content of EVs was analyzed by mass spectrometry. The effects of PDAC-derived EVs on natural kill (NK) cells were investigated by flow cytometry. The serum EVs' TGF-ß1 levels were quantified by ELISA. We found that integrins were enriched in PDAC-derived EVs. The expression of NKG2D, CD107a, TNF-α, and INF-γ in NK cells was significantly downregulated after co-culture with EVs. NK cells also exhibited decreased levels of CD71 and CD98, as well as impaired glucose uptake ability. In addition, NK cell cytotoxicity against pancreatic cancer stem cells was attenuated. Moreover, PDAC-derived EVs induced the phosphorylation of Smad2/3 in NK cells. Serum EVs' TGF-ß1 was significantly increased in PDAC patients. Our findings emphasize the immunosuppressive role of PDAC-derived EVs and provide new insights into our understanding of NK cell dysfunction regarding pre-metastatic niche formation in PDAC.

Kyphoplasty of Osteoporotic Fractured Vertebrae: A Finite Element Analysis about Two Types of Cement.

If conservative treatment of osteoporotic vertebral compression fractures fails, vertebro- or kyphoplasty is indicated. Usually, polymethylmethacrylate cement (PMMA) is applied coming along with many disadvantageous features. Aluminum-free glass-polyalkenoate cement (GPC) appears to be a benefit alternative material. This study aimed at comparing the mean stress values in human vertebrae after kyphoplasty with PMMA and GPC (IlluminOss™) at hand of a finite element analysis. Three models were created performing kyphoplasty using PMMA or IlluminOss™, respectively, at two native, human lumbar vertebrae (L4) while one remains intact. Finite element analysis was performed using CT-scans of every vertebra. Moreover the PMMA-treated vertebra was used as a model as analyses were executed using material data of PMMA and of GPC. The unimpaired, spongious bone showed potentials of 0.25 MPa maximally. After augmentation stress levels showed fivefold increase, rising from externally to internally, revealing stress peaks at the ventral border of the spinal canal. At central areas of cement 1 MPa is measured in both types of cement. Around these central areas the von Mises stress decreased about 25-50% (0.5-0.75 MPa). If workload of 500 N was applied, the stress appeared to be more centralized at the IlluminOss™-model, similar to the unimpaired. Considering the endplates the GPC model also closely resembles the unimpaired. Comparing the PMMA-treated vertebral body and the GPC-simulation, there is an obvious difference. While the PMMA-treated model showed a central stress peak of 5 MPa, the GPC-simulation of the same vertebral body presents lower stress of 1.2-2.5 MPa. Finite element analysis showed that IlluminOss™ (GPC), used in kyphoplasty of vertebral bodies, creates lower level stress and strain compared to standardly used PMMA, leading to lower stress concentrations on the cranial and caudal vertebral surface especially. GPC appears to own advantageous biological and clinical relevant features.

Insertional anatomy of the anterior medial collateral ligament on the sublime tubercle of the elbow.

BACKGROUND: Acute injuries to the anterior medial collateral ligament (AMCL) can occur due to valgus trauma or during other dislocating events to the elbow. AMCL lesions are often associated with bony lesions, such as radial head fractures or fractures of the coronoid process. We analyzed the insertion of the AMCL on the sublime tubercle in relation to surrounding osseous structures. We aimed to increase the understanding of the involvement of the AMCL in bony lesions to the sublime tubercle. METHODS: We investigated 86 elbows from 43 embalmed human specimens. We measured the most ventral extensions of the AMCL at the sublime tubercle in relation to a clearly defined and reproducible landmark. We used as our landmark a horizontal line (baseline) originating on the lesser sigmoid notch in a right angle to the ulnar ridge. RESULTS: The mean distance of the coronoid process tip to the baseline was 4.0 mm (standard deviation [SD], 1.3 mm; range, 1.4-6.7 mm). The mean distance of the ventral extension of the AMCL to the horizontal line was 3.7 mm (SD, 2.6 mm; range: 9.4-2.2 mm). The mean horizontal distance between the ventral aspect of the AMCL and the coronoid tip was 13.7 mm (SD, 2.5 mm; range, 7.7-20.5 mm). CONCLUSIONS: We present a detailed description of the insertional anatomy of the AMCL at the sublime tubercle. These values could be helpful for classifications of coronoid fractures and to estimate the involvement of the AMCL in fractures of the sublime tubercle.

The radial head size in relation to osseous landmarks of the forearm.

PURPOSE: Radial head fractures are regularly treated with radial head arthroplasty. To prevent limited motion or pain, the implant's size should match its normal anatomy. Preoperative estimation of the radial head size helps in finding the correct head component. The aim of this study was to measure bony landmarks in proximity to the radial head to estimate the required size of a prosthesis preoperatively. METHODS: Anatomical landmarks on 82 elbows from 41 embalmed specimens (19 male, 22 female) were measured using a digital caliper after removal of the specimens' tissue: the largest and smallest radial head diameter, length of the radius (styloid tip to radial head articular surface), and the length of the ulna (styloid tip to coronoid base). Additionally, cranio-caudal and antero-posterior diameters of the capitulum on scaled lateral elbow X-ray images were measured. RESULTS: The mean largest and smallest radial head diameters were 24.2 mm (± 2.2, range 19.9-30.3; ICC = 0.992) and 22.5 mm (± 2.0, range 18.9-27.5; ICC = 0.985). The mean radius length was 23.8 cm (± 1.6, range 20.1-27.1; ICC = 0.986), and the mean ulna length was 23.1 cm (± 1.6, range 19.3-26.3; ICC = 0.969). The mean antero-posterior capitulum diameter was 16.2 mm (± 2.4, range 10.4-21.0; ICC = 0.506), and the mean cranio-caudal diameter was 17.0 mm (± 3.3, range 10.0-23.9; ICC = 0.529). The highest correlation to radial head diameters could be shown for diameters of the contralateral radial head and the radius length. CONCLUSIONS: For preoperative estimation of the radial head, the diameters of the contralateral radial head or the radius length are the most accurate.

The ß2 Integrin Mac-1 Induces Protective LC3-Associated Phagocytosis of Listeria monocytogenes.

The intracellular pathogen Listeria monocytogenes (L.m.) is targeted by the autophagic machinery, but the molecular mechanisms involved and consequences for anti-listerial immunity remain enigmatic. Here, we demonstrate that L.m. infection of macrophages in vivo exclusively evokes LC3-associated phagocytosis (LAP), but not canonical autophagy, and that targeting of L.m. by LAP is required for anti-listerial immunity. The pathway leading to LAP induction in response to L.m. infection emanates from the ß2 integrin Mac-1 (CR3, integrin αMß2), a receptor recognizing diverse microbial ligands. Interaction of L.m. with Mac-1 induces acid sphingomyelinase-mediated changes in membrane lipid composition that facilitate assembly and activation of the phagocyte NAPDH oxidase Nox2. Nox2-derived reactive oxygen species then trigger LC3 recruitment to L.m.-containing phagosomes by LAP. By promoting fusion of L.m.-containing phagosomes with lysosomes, LAP increases exposure of L.m. to bactericidal acid hydrolases, thereby enhancing anti-listerial activity of macrophages and immunity of mice.

Depletion of Mitofusin-2 Causes Mitochondrial Damage in Cisplatin-Induced Neuropathy.

Sensory neuropathy is a relevant side effect of the antineoplastic agent cisplatin. Mitochondrial damage is assumed to play a critical role in cisplatin-induced peripheral neuropathy, but the pathomechanisms underlying cisplatin-induced mitotoxicity and neurodegeneration are incompletely understood. In an animal model of cisplatin-induced neuropathy, we determined in detail the extent and spatial distribution of mitochondrial damage during cisplatin treatment. Changes in the total number of axonal mitochondria during cisplatin treatment were assessed in intercostal nerves from transgenic mice that express cyan fluorescent protein. Further, we explored the impact of cisplatin on the expression of nuclear encoded molecules of mitochondrial fusion and fission, including mitofusin-2 (MFN2), optic atrophy 1 (OPA1), and dynamin-related protein 1 (DRP1). Cisplatin treatment resulted in a loss of total mitochondrial mass in axons and in an abnormal mitochondrial morphology including atypical enlargement, increased vacuolization, and loss of cristae. These changes were observed in distal and proximal nerve segments and were more prominent in axons than in Schwann cells. Transcripts of fusion and fission proteins were reduced in distal nerve segments. Significant reduced expression levels of the fusion protein MFN2 was detected in nerves of cisplatin-exposed animals. In summary, we provide for the first time an evidence that cisplatin alters mitochondrial dynamics in peripheral nerves. Loss of MFN2, previously implicated in the pathogenesis of other neurodegenerative diseases, also contributes to the pathogenesis in cisplatin-induced neuropathy.

Cav2.3 (R-Type) Calcium Channels are Critical for Mediating Anticonvulsive and Neuroprotective Properties of Lamotrigine In Vivo.

BACKGROUND/AIMS: Lamotrigine (LTG) is a popular modern antiepileptic drug (AED), however, its mechanism of action has yet to be fully understood, as it is known to modulate many members of several ion channel families. In heterologous systems, LTG inhibits Cav2.3 (R-type) calcium currents, which contribute to kainic-acid- (KA) induced epilepsy in vivo. To gain insight into the role of R-type currents in LTG drug action in vivo, we compared the effects of LTG to topiramate and lacosamide in Cav2.3-deficient mice and controls on KA-induced seizures. METHODS: Behavioral seizure rating and quantitative electrocorticography were performed after injection of 20 mg/kg [and 30 mg/kg] KA. One hour before KA injection, mice were pretreated with either 30 mg/kg LTG, 50 mg/kg topiramate (TPM) or 30 mg/kg lacosamide (LSM). RESULTS: Ablation of Cav2.3 reduced total seizure scores by 28.6% (p=0.0012) and pretreatment with LTG reduced seizure activity of control mice by 23.2% (p=0.02). In Cav2.3-deficient mice LTG pretreatment increased seizure activity by 22.1% (p=0.018) and increased the percentage of degenerated CA1 pyramidal neurons (p=0.02). All three tested AEDs reduced seizure activity in control mice, however only the non-calcium channel modulating AED, LSM had an anticonvulsive effect in Cav2.3-deficient mice. Furthermore LTG altered electrocorticographic parameters differently in the two genotypes, decreasing relative power of ictal spikes in control mice compared to Cav2.3-defcient mice. CONCLUSION: These findings give first in vivo evidence for an essential role for Cav2.3 in LTG pharmacology and shed light on a paradoxical effect of LTG in their absence. Furthermore, LTG appears to promote ictal activity in Cav2.3-deficient mice resulting in increased neurotoxicity in the CA1 region. This paradoxical mechanism, possibly reflecting rebound hyperexcitation of pyramidal CA1 neurons after increased inhibition, may be key in understanding LTG-induced seizure aggravation, observed in clinical practice.

Kinesin-5 Blocker Monastrol Protects Against Bortezomib-Induced Peripheral Neurotoxicity.

Neurotoxicity is a relevant side effect of bortezomib treatment. Previous reports have shown that the development of peripheral neuropathy caused by anti-neoplastic agents may be a result of reduced axonal transport. Based on evidence from prior studies that the kinesin-5 inhibitor monastrol enhances axonal transport and improves neuronal regeneration, we focused on the neuroprotective role of monastrol during the chemotherapeutic treatment with bortezomib. Prolonged treatment of C57BL/6 mice with bortezomib induced a length-dependent small-fiber neuropathy with axonal atrophy and loss of sensory nerve fibers. The administration of monastrol substantially alleviated morphological features of axonal injury and functional measures of sensory neuropathy. Cytotoxicity studies in leukemia and multiple myeloma cell lines showed no interference of monastrol with the cytostatic effects of bortezomib. Our data indicate that the novel approach of targeting microtubule turnover by monastrol provides protection against bortezomib-induced neurotoxicity. The favorable cytotoxic profile of monastrol makes it an interesting candidate as neuroprotective agent in combined chemotherapy regimens that warrants further consideration.

Radial shortening osteotomy reduces radiocapitellar contact pressures while preserving valgus stability of the elbow.

PURPOSE: Shortening osteotomy of the proximal radius might represent a potential salvage procedure in symptomatic radiocapitellar osteoarthritis, which could decrease radiocapitellar load while preserving the native radial head. In an in-vitro biomechanical investigation, we sought to determine whether shortening osteotomy of the proximal radius (1) decreases the radiocapitellar joint pressure upon axial loading and (2) retains valgus stability of the elbow. In addition, the anatomic configuration of the lesser sigmoid notch was evaluated to assess possible contraindications. METHODS: Axial loading (0-400 N) and valgus torque (7.5 N m) over the full range of motion were applied to 14 fresh-frozen specimens before and after shortening osteotomy of the proximal radius by 2.5 mm. Radiocapitellar and ulnohumeral load distribution during axial compression was evaluated using a digital pressure mapping sensor. Valgus displacement was analyzed with a 3D camera system. The inclination angle (α) of the lesser sigmoid notch was assessed via 50 CT scans. RESULTS: Up to axial loading of 250 N, shortening osteotomy caused a significant decrease in radiocapitellar contact pressures (p < 0.041). Valgus stability of specimens did not differ before and after shortening osteotomy (n.s.). The mean inclination angle (α) of the lesser sigmoid notch was 11.3° ± 6.3°. 46% had an inclination angle of ≤ 10° (type I). 46% had an inclination angle of 11°-20° (type II). In 8%, the inclination angle was >20° (type III). CONCLUSION: Shortening osteotomy of the proximal radius can decrease radiocapitellar contact pressures during axial loading of up to 250 N. Primary valgus stability is not relevantly influenced by this procedure. In few patients, shortening osteotomy may cause radioulnar impingement of the radial head at the distal edge of the lesser sigmoid notch due to an inclination angle of >20°. Shortening osteotomy might be a promising treatment option to decrease pain levels in case of isolated radiocapitellar osteoarthritis.

Biomechanical comparison of transosseous re-fixation of the deep fibres of the distal radioulnar ligaments versus deep and superficial fibres: a cadaver study.

PURPOSE: We hypothesized that the re-fixation of the deep and superficial fibres of the distal radioulnar ligaments provide improved stability compared to reconstruction of the deep fibres alone. METHODS: Fourteen fresh-frozen cadaver upper extremities were used for biomechanical testing. Transosseous re-fixation of the deep fibres of the distal radioulnar ligaments alone (single mattress suture group; n = 7) was compared to the transosseous re-attachment of the deep and superficial fibres (double mattress suture group; n = 7). Cyclic load application provoked palmar translation of the radius with respect to the rigidly affixed ulna. Creep, stiffness, and hysteresis were obtained from the load-deformation curves, respectively. Testing was done in neutral forearm rotation, 60° pronation, and 60° supination. RESULTS: The re-fixation techniques did not differ significantly regarding the viscoelastic parameters creep, hysteresis, and stiffness. Several significant differences of one cycle to the consecutive one within each re-fixation group could be detected especially for creep and hysteresis. No significant differences between the different forearm positions could be detected for each viscoelastic parameter. CONCLUSIONS: The re-fixation techniques did not differ significantly regarding creep, hysteresis, and stiffness. This means that the additional re-attachment of the superficial fibres may not provide greater stability to the DRUJ. Bearing in mind that the study was a cadaver examination with a limited number of specimens we may suppose that the re-attachment of the superficial fibres seem to be unnecessary. A gradual decline of creep and hysteresis from first to last loading-unloading cycle is to be expected and typical of ligaments which are viscoelastic.

Paclitaxel inhibits mRNA transport in axons.

Paclitaxel is an integral component of solid tumor treatment. This chemotherapeutic agent provokes an often irreversible peripheral sensory neuropathy with pathological features of distal axonal degeneration. Current pathological concepts assume that polymerization of axonal microtubules and mitochondrial dysfunction contributes to the development of paclitaxel-induced peripheral neuropathy. The relationship, however, between microtubule stabilization, mitotoxicity and axonal degeneration is still not completely understood. To explore the function of axonal mitochondria we treated transgenic mice that harbor cyan fluorescent protein (CFP)-labeled neuronal mitochondria with repeated doses of paclitaxel and assessed neuropathic changes by nerve conduction and histological studies. In addition, mitochondrial content and morphology was determined by ex vivo imaging of axons containing CFP-labeled mitochondria. Using quantitative RT-PCR and fluorescence-labeled mRNA we determined axonal mRNA transport of nuclear encoded mitochondrial proteins. Prolonged treatment with high doses of paclitaxel-induced a predominant sensory neuropathy in mice. Although mitochondrial velocity in axons per se was not altered, we observed significant changes in mitochondrial morphology, suggesting that paclitaxel treatment impairs the dynamics of axonal mitochondria. These changes were caused by decreased levels of nuclear encoded mRNA, including the mitochondrial fusion/fission machinery. Moreover, impaired axonal mRNA transport in vitro resulted in mitochondrial dysfunction and subsequent axonal degeneration. Taken together, our experiments provide evidence that disrupted axonal transport of nuclear derived mRNA plays a crucial role in the pathogenesis of paclitaxel-induced sensory neuropathy.

The course of the median and radial nerve across the elbow: an anatomic study.

INTRODUCTION: Nerve transection has been described as complication of arthroscopic elbow arthrolysis. Therefore, the goal of this study was to define bony landmarks for intraoperative orientation regarding the location of the median and radial nerve. METHODS: In 22 formalin-fixated upper extremities, the radial and median nerves were dissected and marked with respect to their native course. A 3D X-ray scan was performed. The distances of the radial nerve to the radial head (R1), the capitulum (R2), and its lateral border (RC) were measured. The location of the radial nerve in relation to the transversal diameter of the humeral condyle (HC) was calculated. Similarly, the distances of the median nerve to the trochlea (M1), the medial border of the trochlea (M2), and its relation to HC were calculated. RESULTS: The mean value for R1 was 8 mm (±2.9 mm), for R2 was 11.3 mm (±3.8 mm), and for RC was 10.6 mm (±5.1 mm). RC/HC averaged 24 % (±11 %). M1 averaged 11.7 mm (±5.2 mm), and M2 was 2.4 mm (±4.1 mm). M2/HC averaged 6 % (±9 %). CONCLUSIONS: The radial nerve is located ventral to the central third of the capitulum. The median nerve lies ventral to the medial quarter of the humeral condyle. When performing arthroscopic arthrolysis, this information should be kept in mind during anterior capsulectomy.

Susceptibility of murine induced pluripotent stem cell-derived cardiomyocytes to hypoxia and nutrient deprivation.

INTRODUCTION: Induced pluripotent stem cell-derived cardiomyocytes (iPS-CMs) may be suitable for myocardial repair. While their functional and structural properties have been extensively investigated, their response to ischemia-like conditions has not yet been clearly defined. METHODS: iPS-CMs were differentiated and enriched from murine induced pluripotent stem cells expressing enhanced green fluorescent protein (eGFP) and puromycin resistance genes under the control of an α-myosin heavy chain (α-MHC) promoter. iPS-CMs maturity and function were characterized by microscopy, real-time PCR, calcium transient recordings, electrophysiology, and mitochondrial function assays, and compared to those from neonatal murine cardiomyocytes. iPS-CMs as well as neonatal murine cardiomyocytes were exposed for 3 hours to hypoxia (1% O2) and glucose/serum deprivation, and viability, apoptosis markers, reactive oxygen species, mitochondrial membrane potential and intracellular stress signaling cascades were investigated. Then, the iPS-CMs response to mesenchymal stromal cell-conditioned medium was determined. RESULTS: iPS-CMs displayed key morphological and functional properties that were comparable to those of neonatal cardiomyocytes, but several parameters indicated an earlier iPS-CMs maturation stage. During hypoxia and glucose/serum deprivation, iPS-CMs exhibited a significantly higher proportion of poly-caspase-active, 7-aminoactinomycin D-positive and TUNEL-positive cells than neonatal cardiomyocytes. The average mitochondrial membrane potential was reduced in "ischemic" iPS-CMs but remained unchanged in neonatal cardiomyocytes; reactive oxygen species production was only increased in "ischemic" iPS-CMs, and oxidoreductase activity in iPS-CMs dropped more rapidly than in neonatal cardiomyocytes. In iPS-CMs, hypoxia and glucose/serum deprivation led to upregulation of Hsp70 transcripts and decreased STAT3 phosphorylation and total PKCε protein expression. Treatment with mesenchymal stromal cell-conditioned medium preserved oxidoreductase activity and restored pSTAT3 and PKCε levels. CONCLUSION: iPS-CMs appear to be particularly sensitive to hypoxia and nutrient deprivation. Counteracting the ischemic susceptibility of iPS-CMs with mesenchymal stromal cell-conditioned medium may help enhance their survival and efficacy in cell-based approaches for myocardial repair.