Gait analysis in normal and spinal contused mice using the TreadScan system

Advances in spinal cord injury (SCI) research are dependent on quality animal models, which in turn rely on sensitive outcome measures able to detect functional differences in animals following injury. To date, most measurements of dysfunction following SCI rely either on the subjective rating of observers or the slow throughput of manual gait assessment. The present study compares the gait of normal and contusion-injured mice using the TreadScan system. TreadScan utilizes a transparent treadmill belt and a high-speed camera to capture the footprints of animals and automatically analyze gait characteristics. Adult female C57Bl/6 mice were introduced to the treadmill prior to receiving either a standardized mild, moderate, or sham contusion spinal cord injury. TreadScan gait analyses were performed weekly for 10 weeks and compared with scores on the Basso Mouse Scale (BMS). Results indicate that this software successfully differentiates sham animals from injured animals on a number of gait characteristics, including hindlimb swing time, stride length, toe spread, and track width. Differences were found between mild and moderate contusion injuries, indicating a high degree of sensitivity within the system. Rear track width, a measure of the animal's hindlimb base of support, correlated strongly both with spared white matter percentage and with terminal BMS. TreadScan allows for an objective and rapid behavioral assessment of locomotor function following mild-moderate contusive SCI, where the majority of mice still exhibit hindlimb weight support and plantar paw placement during stepping.

Cardiac c-kit+AT2+ cell population is increased in response to ischemic injury and supports cardiomyocyte performance

The expression pattern of angiotensin AT2 receptors with predominance during fetal life and upregulation under pathological conditions during tissue injury/repair process suggests that AT2 receptors may exert an important action in injury/repair adaptive mechanisms. Less is known about AT2 receptors in acute ischemia-induced cardiac injury. We aimed here to elucidate the role of AT2 receptors after acute myocardial infarction. Double immunofluorescence staining showed that cardiac AT2 receptors were mainly detected in clusters of small c-kit+ cells accumulating in peri-infarct zone and c-kit+AT2+ cells increased in response to acute cardiac injury. Further, we isolated cardiac c-kit+AT2+ cell population by modified magnetic activated cell sorting and fluorescence activated cell sorting. These cardiac c-kit+AT2+ cells, represented approximately 0.19% of total cardiac cells in infarcted heart, were characterized by upregulated transcription factors implicated in cardiogenic differentiation (Gata-4, Notch-2, Nkx-2.5) and genes required for self-renewal (Tbx-3, c-Myc, Akt). When adult cardiomyocytes and cardiac c-kit+AT2+ cells isolated from infarcted rat hearts were cocultured, AT2 receptor stimulation in vitro inhibited apoptosis of these cocultured cardiomyocytes. Moreover, in vivo AT2 receptor stimulation led to an increased c-kit+AT2+ cell population in the infarcted myocardium and reduced apoptosis of cardiomyocytes in rats with acute myocardial infarction. These data suggest that cardiac c-kit+AT2+ cell population exists and increases after acute ischemic injury. AT2 receptor activation supports performance of cardiomyocytes, thus contributing to cardioprotection via cardiac c-kit+AT2+ cell population.

Non-selective cutaneous sensory neurectomy as an alternative treatment for auto-mutilation lesion following arthrodesis in three dogs

OBJECTIVE: To describe an alternative method for the treatment of non-responsive self-mutilation injuries in three dogs after carpal/tarsal arthrodesis. STUDY DESIGN: Case series ANIMALS: Two dogs with carpal injury and one dog with tarsal injury treated by arthrodesis METHODS: All dogs developed self-mutilation injuries due to licking and/or chewing of the toes within 21-52 days of surgery. Clinical signs did not resolve within one week after conservative treatment with wound debridement and protective bandages. Following general anaesthesia, a deep horseshoe-shaped skin incision, including the subdermal tissue, was performed proximal to the self-mutilation injury transecting the sensory cutaneous afferent nerves. The skin incision was closed with simple interrupted sutures. RESULTS: All wounds healed without complication. Self-mutilation resolved completely within 24 hours after surgery in all dogs. No recurrence was observed (5 months to 3 years). CONCLUSION: Non-selective cutaneous sensory neurectomy may lead to resolution of self-mutilation following arthrodesis in dogs. CLINICAL RELEVANCE: Failure of conservative treatment in self-mutilation injuries often leads to toe or limb amputation as a last resort. The technique described in this case series is a simple procedure that should be considered prior to amputation. The outcome of this procedure in dogs self-multilating due to neurological or behavioral disturbances unrelated to carpal or tarsal arthrodesis is not known.

Estrogen receptor subtype beta2 is involved in neuromast development in zebrafish (Danio rerio) larvae

Estrogens are known to play a role in both reproductive and non-reproductive functions in mammals. Estrogens and their receptors are involved in the development of the central nervous system (brain development, neuronal survival and differentiation) as well as in the development of the peripheral nervous system (sensory-motor behaviors). In order to decipher possible functions of estrogens in early development of the zebrafish sensory system, we investigated the role of estrogen receptor beta(2) (ERbeta(2)) by using a morpholino (MO) approach blocking erbeta(2) RNA translation. We further investigated the development of lateral line organs by cell-specific labeling, which revealed a disrupted development of neuromasts in morphants. The supporting cells developed and migrated normally. Sensory hair cells, however, were absent in morphants' neuromasts. Microarray analysis and subsequent in situ hybridizations indicated an aberrant activation of the Notch signaling pathway in ERbeta(2) morphants. We conclude that signaling via ERbeta(2) is essential for hair cell development and may involve an interaction with the Notch signaling pathway during cell fate decision in the neuromast maturation process.

Histological evaluation of human intrabony periodontal defects treated with an unsintered nanocrystalline hydroxyapatite paste

OBJECTIVES The aim of the study was to clinically and histologically evaluate the healing of human intrabony defects treated with open flap surgery (OFD) and application of a new, resorbable, fully synthetic, unsintered, nanocrystalline, phase-pure hydroxyapatite (nano-HA). MATERIALS AND METHODS Six patients, each of them displaying very advanced intrabony defects around teeth scheduled for extraction due to advanced chronic periodontitis and further prosthodontic considerations, were included in the study. Following local anaesthesia, mucoperiosteal flaps were reflected; the granulation tissue was removed, and the roots were meticulously debrided by hand and ultrasonic instruments. A notch was placed at the most apical extent of the calculus present on the root surface or at the most apical part of the defect (if no calculus was present) in order to serve as a reference for the histological evaluation. Following defect fill with nano-HA, the flaps were sutured by means of mattress sutures to allow primary intention healing. At 7 months after regenerative surgery, the teeth were extracted together with some of their surrounding soft and hard tissues and processed for histological analysis. RESULTS The postoperative healing was uneventful in all cases. At 7 months following surgery, mean PPD reduction and mean CAL gain measured 4.0 ± 0.8 and 2.5 ± 0.8 mm, respectively. The histological analysis revealed a healing predominantly characterized by epithelial downgrowth. Limited formation of new cementum with inserting connective tissue fibers and bone regeneration occurred in three out of the six biopsies (i.e. 0-0.86 and 0-1.33 mm, respectively). Complete resorption of the nano-HA was found in four out of the six biopsies. A few remnants of the graft particles (either surrounded by newly formed mineralized tissue or encapsulated in connective tissue) were found in two out of the six biopsies. CONCLUSION Within their limits, the present results indicate that nano-HA has limited potential to promote periodontal regeneration in human intrabony defects. CLINICAL RELEVANCE The clinical outcomes obtained following surgery with OFD + nano-HA may not reflect true periodontal regeneration.

The prevalence of widespread central hypersensitivity in chronic pain patients

BACKGROUND Chronic pain is associated with generalized hypersensitivity and impaired endogenous pain modulation (conditioned pain modulation; CPM). Despite extensive research, their prevalence in chronic pain patients is unknown. This study investigated the prevalence and potential determinants of widespread central hypersensitivity and described the distribution of CPM in chronic pain patients. METHODS We examined 464 consecutive chronic pain patients for generalized hypersensitivity and CPM using pressure algometry at the second toe and cold pressor test. Potential determinants of generalized central hypersensitivity were studied using uni- and multivariate regression analyses. Prevalence of generalized central hypersensitivity was calculated for the 5th, 10th and 25th percentile of normative values for pressure algometry obtained by a previous large study on healthy volunteers. CPM was addressed on a descriptive basis, since normative values are not available. RESULTS Depending on the percentile of normative values considered, generalized central hypersensitivity affected 17.5-35.3% of patients. 23.7% of patients showed no increase in pressure pain threshold after cold pressor test. Generalized central hypersensitivity was more frequent and CPM less effective in women than in men. Unclearly classifiable pain syndromes showed higher frequencies of generalized central hypersensitivity than other pain syndromes. CONCLUSIONS Although prevalent in chronic pain, generalized central hypersensitivity is not present in every patient. An individual assessment is therefore required in order to detect altered pain processing. The broad basic knowledge about central hypersensitivity now needs to be translated into concrete clinical consequences, so that patients can be offered an individually tailored mechanism-based treatment.

Distal radius attachments of the radiocarpal ligaments: an anatomical study

Background Understanding the anatomy of the ligaments of the distal radius aids in the surgical repair of ligamentous injuries and the prediction of intraarticular fracture patterns. Purposes (1) to measure the horizontal and vertical distances of the origins of the radiocarpal ligaments from the most ulnar corner of the sigmoid notch and the joint line, respectively; and (2) to express them as a percentile of the total width of the bony distal radius. Methods We dissected 8 cadaveric specimens and identified the dorsal radiocarpal, radioscaphocapitate, and the long and short radiolunate ligaments. Results The dorsal radiocarpal ligament attached from the 16th to the 52nd percentile of the radial width. The radioscaphocapitate ligament attached around the radial styloid from the 86th percentile volarly to the 87th percentile dorsally. The long radiolunate ligament attached from the 59th to the 85th percentile, and the short radiolunate ligament attached from the 14th to the 41st percentile. Discussion There was a positive correlation between the radial width and the horizontal distance of the ligaments from the sigmoid notch. These findings may aid individualized surgical repair or reconstruction adjusted to patient size and enable further standardized research on distal radial fractures and their relationship with radiocarpal ligaments.

Lameness and foot lesions in Swiss dairy cows: I. Prevalence.

Prevalences of foot lesions and lameness were recorded in 1'449 Swiss dairy cows during routine claw-trimming on 78 farms from June 2010 until February 2011. Lameness was present in 14.8 % of cows and on 80.8 % of investigated farms. Highest prevalences were seen for widened white line (80.7 %/100 %), signalling foot lesion (65.6 %/98.7 %), heel-horn erosion (34.2 %/88.5 %), digital dermatitis complex (29.1 %/73.1 %), severe hemorrhages (27.9 %/87.2 %), and Rusterholz' sole ulcers (11.5 %/74.4 %) at cow and herd level, respectively. Lower prevalences were found for subclinical laminitis (5.4 %/47.4 %), chronic laminitis (3.3 %/25.6 %), white line disease (4.7 %/42.3 %), double soles (2.6 %/33.3 %), interdigital hyperplasia (3.1 %/33.3 %), sole ulcers (0.4 %/6.4 %), toe infections caused by faulty claw-trimming (3.9 %/39.7 %) and by injury (0.1 %/2.6 %), deep lacerations (0.4 %/6.4 %), and interdigital phlegmona (0.1 %/1.3 %). Lameness and foot lesions were shown to represent important health problems of dairy cows under the conditions of the typical grass-based production system in Switzerland. Digital dermatitis has developed to the most relevant foot disease with a high impact on welfare of Swiss dairy cows within the past 10 years.

Delta like ligand 4 is a critical regulator of bone marrow cell differentiation into pericytes/vascular smooth muscle cells and is essential for the vasculogenesis that supports the growth of Ewing’s sarcoma

We have previously shown that vasculogenesis, the process by which bone marrow-derived cells are recruited to the tumor and organized to form a blood vessel network de novo, is essential for the growth of Ewing’s sarcoma. We further demonstrated that these bone marrow cells differentiate into pericytes/vascular smooth muscle cells(vSMC) and contribute to the formation of the functional vascular network. The molecular mechanisms that control bone marrow cell differentiation into pericytes/vSMC in Ewing’s sarcoma are poorly understood. Here, we demonstrate that the Notch ligand Delta like ligand 4 (DLL4) plays a critical role in this process. DLL4 is essential for the formation of mature blood vessels during development and in several tumor models. Inhibition of DLL4 causes increased vascular sprouting, decreased pericyte coverage, and decreased vessel functionality. We demonstrate for the first time that DLL4 is expressed by bone marrow-derived pericytes/vascular smooth muscle cells in two Ewing’s sarcoma xenograft models and by perivascular cells in 12 out of 14 patient samples. Using dominant negative mastermind to inhibit Notch, we demonstrate that Notch signaling is essential for bone marrow cell participation in vasculogenesis. Further, inhibition of DLL4 using either shRNA or the monoclonal DLL4 neutralizing antibody YW152F led to dramatic changes in blood vessel morphology and function. Vessels in tumors where DLL4 was inhibited were smaller, lacked lumens, had significantly reduced numbers of bone marrow-derived pericyte/vascular smooth muscle cells, and were less functional. Importantly, growth of TC71 and A4573 tumors was significantly inhibited by treatment with YW152F. Additionally, we provide in vitro evidence that DLL4-Notch signaling is involved in bone marrow-derived pericyte/vascular smooth muscle cell formation outside of the Ewing’s sarcoma environment. Pericyte/vascular smooth muscle cell marker expression by whole bone marrow cells cultured with mouse embryonic stromal cells was reduced when DLL4 was inhibited by YW152F. For the first time, our findings demonstrate a role for DLL4 in bone marrow-derived pericyte/vascular smooth muscle differentiation as well as a critical role for DLL4 in Ewing’s sarcoma tumor growth.

Robust estimates of climate-induced hydrological change in a temperate mountainous region

A sustainable water resources management depends on sound information about the impacts of climate change. This information is, however, not easily derived because natural runoff variability interferes with the climate change signal. This study presents a procedure that leads to robust estimates of magnitude and Time Of Emergence (TOE) of climate-induced hydrological change that also account for the natural variability contained in the time series. Firstly, natural variability of 189 mesoscale catchments in Switzerland is sampled for 10 ENSEMBLES scenarios for the control (1984–2005) and two scenario periods (near future: 2025–2046, far future: 2074–2095) applying a bootstrap procedure. Then, the sampling distributions of mean monthly runoff are tested for significant differences with the Wilcoxon-Mann–Whitney test and for effect size with Cliff’s delta d. Finally, the TOE of a climate change induced hydrological change is determined when at least eight out of the ten hydrological projections significantly differ from natural variability. The results show that the TOE occurs in the near future period except for high-elevated catchments in late summer. The significant hydrological projections in the near future correspond, however, to only minor runoff changes. In the far future, hydrological change is statistically significant and runoff changes are substantial. Temperature change is the most important factor determining hydrological change in this mountainous region. Therefore, hydrological change depends strongly on a catchment’s mean elevation. Considering that the hydrological changes are predicted to be robust in the near future highlights the importance of accounting for these changes in water resources planning.

Time of emergence of trends in ocean biogeochemistry

For the detection of climate change, not only the magnitude of a trend signal is of significance. An essential issue is the time period required by the trend to be detectable in the first place. An illustrative measure for this is time of emergence (ToE), that is, the point in time when a signal finally emerges from the background noise of natural variability. We investigate the ToE of trend signals in different biogeochemical and physical surface variables utilizing a multi-model ensemble comprising simulations of 17 Earth system models (ESMs). We find that signals in ocean biogeochemical variables emerge on much shorter timescales than the physical variable sea surface temperature (SST). The ToE patterns of pCO2 and pH are spatially very similar to DIC (dissolved inorganic carbon), yet the trends emerge much faster – after roughly 12 yr for the majority of the global ocean area, compared to between 10 and 30 yr for DIC. ToE of 45–90 yr are even larger for SST. In general, the background noise is of higher importance in determining ToE than the strength of the trend signal. In areas with high natural variability, even strong trends both in the physical climate and carbon cycle system are masked by variability over decadal timescales. In contrast to the trend, natural variability is affected by the seasonal cycle. This has important implications for observations, since it implies that intra-annual variability could question the representativeness of irregularly sampled seasonal measurements for the entire year and, thus, the interpretation of observed trends.

Surgical anatomy of the feline sacroiliac joint for lag screw fixation of sacroiliac fracture-luxation

Twenty-eight feline pelves (56 hemipelves) were examined in order to identify the location for optimal sacroiliac screw placement in sacroiliac fracture-luxation repair. A drill hole was started on the median plane of the hemipelvis in the centre of the body of the first sacral segment until it penetrated the lateral cortex of the ilial wing, thus providing optimal drill hole placement. The position of the drill hole on the articular surface of the sacral wing and on the lateral surface of the ilial wing was measured. The distance of the drill hole from the cranial margin of the sacral wing was 51% of sacral wing length, just cranial to the crescent shaped hyaline cartilage. The distance from the dorsal margin was 47% of sacral wing height. The drill bit direction has to be adjusted to the cranio-caudal inclination (range 10° to 29°) and dorso-ventral inclination (range 2° to 25°) of the sacral wing. A notch in the cranial edge of the sacral wing was present, with variable position, in 34% of the specimens and is consequently not a useful landmark for sacroiliac screw placement. The drill hole on the lateral surface of the ilium was located in craniocaudal direction at a distance of 69% of sacral tuber length, measured from the cranial dorsal iliac spine. The dorso-ventral position of the drill hole was at a distance of 52% of ilial wing height measured from the sacral tuber. The ventral gluteal line, present in 93% of the cases, is a useful landmark to locate optimal screw hole position on the ilial wing.

Anatomy, biology, physiology and basic pathology of the nail organ

The nail is the largest skin appendage. It grows continuously through life in a non-cyclical manner; its growth is not hormone-dependent. The nail of the middle finger of the dominant hand grows fastest with approximately 0.1 mm/day, whereas the big toe nail grows only 0.03-0.05 mm/d. The nails' size and shape vary characteristically from finger to finger and from toe to toe, for which the size and shape of the bone of the terminal phalanx is responsible. The nail apparatus consists of both epithelial and connective tissue components. The matrix epithelium is responsible for the production of the nail plate whereas the nail bed epithelium mediates firm attachment. The hyponychium is a specialized structure sealing the subungual space and allowing the nail plate to physiologically detach from the nail bed. The proximal nail fold covers most of the matrix. Its free end forms the cuticle which seals the nail pocket or cul-de-sac. The dermis of the matrix and nail bed is specialized with a morphogenetic potency. The proximal and lateral nail folds form a frame on three sides giving the nail stability and allowing it to grow out. The nail protects the distal phalanx, is an extremely versatile tool for defense and dexterity and increases the sensitivity of the tip of the finger. Nail apparatus, finger tip, tendons and ligaments of the distal interphalangeal joint form a functional unit and cannot be seen independently. The nail organ has only a certain number of reaction patterns that differ in many respects from hairy and palmoplantar skin.

Reliability of Quantitative Sensory Tests in a Low Back Pain Population.

BACKGROUND AND OBJECTIVES Reliability is an essential condition for using quantitative sensory tests (QSTs) in research and clinical practice, but information on reliability in patients with chronic pain is sparse. The aim of this study was to evaluate the reliability of different QST in patients with chronic low back pain. METHODS Eighty-nine patients with chronic low back pain participated in 2 identical experimental sessions, separated by at least 7 days. The following parameters were recorded: pressure pain detection and tolerance thresholds at the toe, electrical pain thresholds to single and repeated stimulation, heat pain detection and tolerance thresholds at the arm and leg, cold pain detection threshold at the arm and leg, and conditioned pain modulation using the cold pressor test.Reliability was analyzed using the coefficient of variation, the coefficient of repeatability, and the intraclass correlation coefficient. It was judged as acceptable or not based primarily on the analysis of the coefficient of repeatability. RESULTS The reliability of most tests was acceptable. Exceptions were cold pain detection thresholds at the leg and arm. CONCLUSIONS Most QST measurements have acceptable reliability in patients with chronic low back pain.

Clinical Significance of NOTCH1 and NOTCH2 Expression in Gastric Carcinomas: An Immunohistochemical Study.

BACKGROUND NOTCH signaling can exert oncogenic or tumor suppressive functions and can contribute to chemotherapy resistance in cancer. In this study, we aimed to clarify the clinicopathological significance and the prognostic and predictive value of NOTCH1 and NOTCH2 expression in gastric cancer (GC). METHODS NOTCH1 and NOTCH2 expression was determined immunohistochemically in 142 primarily resected GCs using tissue microarrays and in 84 pretherapeutic biopsies from patients treated by neoadjuvant chemotherapy. The results were correlated with survival, response to therapy, and clinico-pathological features. RESULTS Primarily resected patients with NOTCH1-negative tumors demonstrated worse survival. High NOTCH1 expression was associated with early-stage tumors and with significantly increased survival in this subgroup. Higher NOTCH2 expression was associated with early-stage and intestinal-type tumors and with better survival in the subgroup of intestinal-type tumors. In pretherapeutic biopsies, higher NOTCH1 and NOTCH2 expression was more frequent in non-responding patients, but these differences were statistically not significant. CONCLUSION Our findings suggested that, in particular, NOTCH1 expression indicated good prognosis in GC. The close relationship of high NOTCH1 and NOTCH2 expression with early tumor stages may indicate a tumor-suppressive role of NOTCH signaling in GC. The role of NOTCH1 and NOTCH2 in neoadjuvantly treated GC is limited.