A Nanometric Probe of the Local Proton Concentration in Microtubule-Based Biophysical Systems.

We show a double-functional fluorescence sensing paradigm that can retrieve nanometric pH information on biological structures. We use this method to measure the extent of protonic condensation around microtubules, which are protein polymers that play many roles crucial to cell function. While microtubules are believed to have a profound impact on the local cytoplasmic pH, this has been hard to show experimentally due to the limitations of conventional sensing techniques. We show that subtle changes in the local electrochemical surroundings cause a double-functional sensor to transform its spectrum, thus allowing a direct measurement of the protonic concentration at the microtubule surface. Microtubules concentrate protons by as much as one unit on the pH scale, indicating a charge storage role within the cell via the localized ionic condensation. These results confirm the bioelectrical significance of microtubules and reveal a sensing concept that can deliver localized biochemical information on intracellular structures.

Modeling Microtubule Counterion Distributions and Conductivity Using the Poisson-Boltzmann Equation.

Microtubules are highly negatively charged proteins which have been shown to behave as bio-nanowires capable of conducting ionic currents. The electrical characteristics of microtubules are highly complicated and have been the subject of previous work; however, the impact of the ionic concentration of the buffer solution on microtubule electrical properties has often been overlooked. In this work we use the non-linear Poisson Boltzmann equation, modified to account for a variable permittivity and a Stern Layer, to calculate counterion concentration profiles as a function of the ionic concentration of the buffer. We find that for low-concentration buffers ([KCl] from 10 µM to 10 mM) the counterion concentration is largely independent of the buffer's ionic concentration, but for physiological-concentration buffers ([KCl] from 100 to 500 mM) the counterion concentration varies dramatically with changes in the buffer's ionic concentration. We then calculate the conductivity of microtubule-counterion complexes, which are found to be more conductive than the buffer when the buffer's ionic concentrations is less than ≈100 mM and less conductive otherwise. These results demonstrate the importance of accounting for the ionic concentration of the buffer when analyzing microtubule electrical properties both under laboratory and physiological conditions. We conclude by calculating the basic electrical parameters of microtubules over a range of ionic buffer concentrations applicable to nanodevice and medical applications.

Revealing and Attenuating the Electrostatic Properties of Tubulin and Its Polymers.

Tubulin is an electrostatically negative protein that forms cylindrical polymers termed microtubules, which are crucial for a variety of intracellular roles. Exploiting the electrostatic behavior of tubulin and microtubules within functional microfluidic and optoelectronic devices is limited due to the lack of understanding of tubulin behavior as a function of solvent composition. This work displays the tunability of tubulin surface charge using dimethyl sulfoxide (DMSO) for the first time. Increasing the DMSO volume fractions leads to the lowering of tubulin's negative surface charge, eventually causing it to become positive in solutions >80% DMSO. As determined by electrophoretic mobility measurements, this change in surface charge is directionally reversible, i.e., permitting control between -1.5 and + 0.2 cm2  (V s)-1 . When usually negative microtubules are exposed to these conditions, the positively charged tubulin forms tubulin sheets and aggregates, as revealed by an electrophoretic transport assay. Fluorescence-based experiments also indicate that tubulin sheets and aggregates colocalize with negatively charged g-C3 N4 sheets while microtubules do not, further verifying the presence of a positive surface charge. This study illustrates that tubulin and its polymers, in addition to being mechanically robust, are also electrically tunable.

All Wired Up: An Exploration of the Electrical Properties of Microtubules and Tubulin.

Microtubules are hollow, cylindrical polymers of the protein α, ß tubulin, that interact mechanochemically with a variety of macromolecules. Due to their mechanically robust nature, microtubules have gained attention as tracks for precisely directed transport of nanomaterials within lab-on-a-chip devices. Primarily due to the unusually negative tail-like C-termini of tubulin, recent work demonstrates that these biopolymers are also involved in a broad spectrum of intracellular electrical signaling. Microtubules and their electrostatic properties are discussed in this Review, followed by an evaluation of how these biopolymers respond mechanically to electrical stimuli, through microtubule migration, electrorotation and C-termini conformation changes. Literature focusing on how microtubules act as nanowires capable of intracellular ionic transport, charge storage, and ionic signal amplification is reviewed, illustrating how these biopolymers attenuate ionic movement in response to electrical stimuli. The Review ends with a discussion on the important questions, challenges, and future opportunities for intracellular microtubule-based electrical signaling.

[Acute iron poisoning : Successful treatment of a potentially fatal suicidal attempt]. / Akute Eisenintoxikation : Erfolgreiche Therapie einer potenziell tödlichen parasuizidalen Geste.

We present the case of a patient who took 150 mg per kg bodyweight of iron in a suicidal attempt. We illustrate the diagnostic and therapeutic procedures required to successfully cope with this poisoning, which is, by surprise, potentially lethal.

[A woman with lower back pain]. / Een vrouw met pijn in de onderrug.

A 88-year old woman presented with severe pain in the lower back which radiated to her left leg. She was diagnosed with a sacral insufficiency fracture. Sacral insufficiency fractures are a cause of lower back pain in the elderly population that is frequently overlooked because the symptoms and signs are not specific.

Cerebral atrophy as outcome measure in short-term phase 2 clinical trials in multiple sclerosis.

INTRODUCTION: Cerebral atrophy is a compound measure of the neurodegenerative component of multiple sclerosis (MS) and a conceivable outcome measure for clinical trials monitoring the effect of neuroprotective agents. In this study, we evaluate the rate of cerebral atrophy in a 6-month period, investigate the predictive and explanatory value of other magnetic resonance imaging (MRI) measures in relation to cerebral atrophy, and determine sample sizes for future short-term clinical trials using cerebral atrophy as primary outcome measure. METHODS: One hundred thirty-five relapsing-remitting multiple sclerosis patients underwent six monthly MRI scans from which the percentage brain volume change (PBVC) and the number and volume of gadolinium (Gd)-enhancing lesions, T2 lesions, and persistent black holes (PBH) were determined. By means of multiple linear regression analysis, the relationship between focal MRI variables and PBVC was assessed. Sample size calculations were performed for all patients and subgroups selected for enhancement or a high T2 lesion load at baseline. RESULTS: A significant atrophy occurred over 6 months (PBVC = -0.33%, SE = 0.061, p < 0.0001). The number of baseline T2 lesions (p = 0.024), the on-study Gd-enhancing lesion volume (p = 0.044), and the number of on-study PBHs (p = 0.003) were associated with an increased rate of atrophy. For a 50% decrease in rate of atrophy, the sample size calculations showed that approximately 283 patients per arm are required in an unselected sampled population and 185 patients per arm are required in a selected population. CONCLUSION: Within a 6-month period, significant atrophy can be detected and on-study associations of PBVC and PBHs emphasizes axonal loss to be a driving mechanism. Application as primary outcome measure in short-term clinical trials with feasible sample size requires a potent drug to obtain sufficient power.

Video analysis of trunk and knee motion during non-contact anterior cruciate ligament injury in female athletes: lateral trunk and knee abduction motion are combined components of the injury mechanism.

BACKGROUND: The combined positioning of the trunk and knee in the coronal and sagittal planes during non-contact anterior cruciate ligament (ACL) injury has not been previously reported. HYPOTHESIS: During ACL injury female athletes demonstrate greater lateral trunk and knee abduction angles than ACL-injured male athletes and uninjured female athletes. DESIGN: Cross-section control-cohort design. METHODS: Analyses of still captures from 23 coronal (10 female and 7 male ACL-injured players and 6 female controls) or 28 sagittal plane videos performing similar landing and cutting tasks. Significance was set at p < or = 0.05. RESULTS: Lateral trunk and knee abduction angles were higher in female compared to male athletes during ACL injury (p < or = 0.05) and trended toward being greater than female controls (p = 0.16, 0.13, respectively). Female ACL-injured athletes showed less forward trunk lean than female controls (mean (SD) initial contact (IC): 1.6 (9.3) degrees vs 14.0 (7.3) degrees, p < or = 0.01). CONCLUSION: Female athletes landed with greater lateral trunk motion and knee abduction during ACL injury than did male athletes or control females during similar landing and cutting tasks. CLINICAL RELEVANCE: Lateral trunk and knee abduction motion are important components of the ACL injury mechanism in female athletes as observed from video evidence of ACL injury.

Carotid stenosis and carotid plaque analysis relevant to carotid endarterectomy and stent-assisted angioplasty.

The primary objective of this cadaveric study was to review the morphological variations of the anatomy of the human carotid artery bifurcation relevant to carotid endarterectomy (CEA) and carotid artery stent-supported angioplasty (CSSA). We quantify carotid bifurcation plaque morphology. Results showed that the angle of deviation at the origin of the internal carotid artery (ICA), in relation to the common carotid artery (CCA), measured a mean of 21.8 degrees with a range from seven to 45 degrees. This anatomical finding is important for the interventionalist concerned with insertion of a carotid stent. The angle of the ICA origin may be an independent risk factor for early atherosclerotic changes at the ICA bulb. Carotid bifurcation plaque was observed in a small, random cohort of seven out of 13 cadavers, and contributed to a mean stenosis of 15.2% (range 5.0-34.8%). Plaque morphology (n = 7) showed haemorrhage (29%), superficial thrombosis (57%), calcification (71%), areas of focal necrosis (71%), neovascularisation (14%) and infiltrates (29%). Ulcerations were not detected. Although four out of 13 patients (31%) died of a cerebrovascular accident, the cause of cerebral apoplexy was thought not to be associated with the carotid bifurcation pathology. 'Re-boring' of occluding plaque, as in CEA, offers potential volumetric anatomical advantage over CSSA within the carotid bifurcation and bulb. In conclusion, precise and applied knowledge of carotid bifurcation anatomy is critical to reduce technical complications during CEA or CSSA. This information may reduce potential dangers of iatrogenic thrombo-embolism and ensuing neurologic deficits. Patients with low-grade carotid stenosis, evidence of focal plaque necrosis, are at risk of spontaneous plaque cap rupture, distal thromboembolism and stroke.

EVAR: critical applied aortic morphology relevant to type-II endoleaks following device enhancement in patients with abdominal aortic aneurysms.

Endovascular repair (EVAR) of abdominal aortic aneurysms (AAA) is an established alternative option to conventional surgery for AAA, provided optimal anatomical morphology of the aneurysm sac, neck and outflow exists. In most documented series of EVAR, type-II endoleak occurrence is a universal procedural drawback. This is referred to as the Achilles heel of EVAR. This morphological study, addressing predominantly non-aneurysmal aortic anatomy, reveals the dyssynchronous origins of the renal ostia, ectopia of the superior mesenteric artery and median sacral artery, variations in the length of the infrarenal abdominal aorta, multiple mainstem renal arteries, and the presence of accessory renal arteries (in 13% of cadavers). Such potential vascular anomalies need careful consideration pre-operatively prior to EVAR. In a prospective, clinical study of EVAR in 163 patients over 60 months, using four different aortic stent devices, we demonstrated an intraprocedural type-II endoleak rate, before exclusion, of 3% (5/163). Most were related to patent lumbar arteries. An active policy of intraprocedural aneurysm pressure sac measurement and angiography was used to demonstrate type-I and type-II endoleaks, focusing on the applied anatomy of aortic side branches and variations. Selective intraprocedural coil embolisation and thrombin injection into the sac was utilised to thrombose persisting and large lumbar arteries that predisposed to retroleaks. We recorded a low incidence of persisting type-II endoleaks using this proactive treatment strategy by addressing variant aortic morphology and patent lumbar arteries during EVAR. One aneurysm-related death (0.6%) was observed due to late rupture after EVAR, and a single intraprocedural death was related to unpredictable aneurysm rupture. In conclusion, comprehensive anatomical knowledge of the abdominal aorta and its main collateral side branches, including variations, is a fundamental prerequisite if satisfactory and predictable results are to be achieved after EVAR, especially regarding prevention, diagnosis and treatment of type-II endoleaks. Intraprocedural aneurysm sac pressure monitoring, coil embolisation and the use of injection of thrombin into the aneurysm sac of selected patients is useful in reducing the incidence of post-EVAR type-II persisting endoleaks.

Low-risk stress fractures.

Stress fractures can occur in almost any bone in the body, with the lower extremity weightbearing bones, especially the tibia, tarsals, and metatarsals, being affected most frequently. Although the cause of these fractures is multifactoral, repetitive physical forces without adequate rest are the primary culprits. Stress fractures may be broadly classified as low-risk or high-risk injuries. Low-risk stress fractures, the topic of this review article, can be diagnosed through a thorough history, physical examination, and radiographs. Nuclear scintigraphy is occasionally necessary for confirmation, especially for fractures of the spine and pelvis. When diagnosed early and treated with restriction of activity, low-risk stress fractures have a favorable prognosis.

Catastrophic injuries in pole-vaulters.

Pole vaulting is a unique sport in that athletes often land from heights ranging from 10 to 20 feet. We retrospectively reviewed 32 catastrophic pole-vault injuries that were reported to the National Center for Catastrophic Sports Injury Research between 1982 and 1998. The purpose of this study was to determine the mechanisms of injury so that preventive strategies can be implemented. Information was obtained by means of a telephone interview with someone familiar with the accident. All injuries occurred in male athletes at an average age of 17.5 years; 31 were catastrophic head injuries and 1 was a thoracic spine fracture that resulted in paraplegia. Three common mechanisms were identified: 17 (53%) athletes landed with their body on the landing pad and their head on the surrounding hard ground, 8 (25%) landed in the vault box after being stranded at the height of the jump, and 5 (16%) completely missed the landing pad. The mechanism of injury in the remaining two athletes was unknown. The accident resulted in death in 16 (50%) athletes and in permanent disability in 6 (19%). Increasing the minimum landing pad size and enforcing the rule requiring soft surfaces adjacent to the landing pads are the primary recommendations for preventing injuries. The authors discuss other rule and equipment changes that may help reduce the occurrence of future injuries.

High-risk stress fractures: evaluation and treatment.

Stress fractures are common overuse injuries seen in athletes and military recruits. The pathogenesis is multifactorial and usually involves repetitive submaximal stresses. Intrinsic factors, such as hormonal imbalances, may also contribute to the onset of stress fractures, especially in women. The classic presentation is a patient who experiences the insidious onset of pain after an abrupt increase in the duration or intensity of exercise. The diagnosis is primarily clinical, but imaging modalities such as plain radiography, scintigraphy, computed tomography, and magnetic resonance imaging may provide confirmation. Most stress fractures are uncomplicated and can be managed by rest and restriction from the precipitating activity. A subset of stress fractures can present a high risk for progression to complete fracture, delayed union, or nonunion. Specific sites for this type of stress fracture are the femoral neck (tension side), the patella, the anterior cortex of the tibia, the medial malleolus, the talus, the tarsal navicular, the fifth metatarsal, and the great toe sesamoids. Tensile forces and the relative avascularity at the site of a stress-induced fracture often lead to poor healing. Therefore, high-risk stress fractures require aggressive treatment.

Rapid cardiopulmonary support for children with complex congenital heart disease.

BACKGROUND: Extracorporeal membrane oxygenation has limitations in children with congenital heart disease (prolonged setup times, increased postoperative blood loss, and difficulty during transport). We developed a miniaturized cardiopulmonary support circuit to address these limitations. PATIENTS AND METHODS: The cardiopulmonary support system includes a preassembled, completely heparin-coated circuit, a BP-50 Bio-Medicus centrifugal pump, a Minimax plus membrane oxygenator, a Bio-Medicus flow probe, and a Bio-trend hematocrit/oxygen saturation monitor. Short tubing length permits a 250-mL bloodless prime in less than 5 minutes. From 1995 to 1997, 23 children with congenital heart disease were supported with this technique. RESULTS: Overall survival to discharge was 48% (11 of 23 patients). Survival to discharge was 80% (4 of 5) in the preoperative support group, 20% (1 of 5) in the postoperative failure to wean from cardiopulmonary bypass group, 44% (4 of 9) in the group placed on support postoperatively after transfer to the intensive care unit, and 50% (2 of 4 patients) in the nonoperative group. Neonatal cardiopulmonary support survival to discharge was 46% (6 of 13 patients). CONCLUSIONS: This pediatric cardiopulmonary support system is safe and effective. Advantages over conventional extracorporeal membrane oxygenation include rapid setup time, decreased postoperative blood loss, and simplified transport.

Mechanisms of anterior cruciate ligament injury.

This study examined the mechanisms of anterior cruciate ligament (ACL) injury. In the first part of the study, using a comprehensive, standardized questionnaire, 89 athletes (100 knees) were interviewed about the events surrounding their ACL injury. A noncontact mechanism was reported in 71 (72%) knees and a contact injury in 28 (28%) knees; one patient was unsure if there was any contact. Most of the injuries were sustained at footstrike with the knee close to full extension. Noncontact mechanisms were classified as sudden deceleration prior to a change of direction or landing motion, while contact injuries occurred as a result of valgus collapse of the knee. Hamstring flexibility parameters revealed a statistically higher level of laxity in the injured athletes compared with a matched group of 28 controls. In the second part of the study, videotapes of 27 separate ACL disruptions were reviewed and confirmed that most noncontact injuries occur with the knee close to extension during a sharp deceleration or landing maneuver. Because the knee is in a position to allow the extensor mechanism to strain the ACL and maximum, eccentric muscle force conditions usually apply, the quadriceps may play an important role in ACL disruption. Passive protection of the ACL by the hamstring muscles may be reduced in patients with above-average flexibility.

Etiology and Prevention of Noncontact ACL Injury.

An understanding of the etiology and prevention of noncontact ACL injuries has lagged behind diagnosis and treatment. However, a growing research implicates hormonal, anatomic, environmental, and neuromuscular factors that may predispose athletes, particularly women, to these injuries. Specific factors may include estrogen levels, the shape of the intercondylar notch, playing style, and neuromuscular control of the quadriceps and hamstring muscles. Prevention programs that involve proprioception, plyometrics, strength training, and improved jumping, stopping, and turning techniques show promising results.

Tibia and fibula fractures in soccer players.

We performed a retrospective review of 31 athletes who sustained a fracture of the lower leg from a direct blow while playing soccer. Fifteen fractures involved both the tibia and fibula 11 only the tibia, and 5 only the fibula. Information was collected using a standardized questionnaire. The mean follow-up from the time of injury was 30 months. Injuries typically occurred in young, competitive athletes during game situations. The mechanisms were broadly classified into several categories: contact during a slide tackle (13, 42%), a collision with the goalkeeper (8, 26%), two opposing players colliding while swinging for a loose ball (7, 23%), or a player being kicked by a standing opponent (3, 10%). The majority of fractures (26, 90%) occurred while the athletes were wearing shin guards. The point of impact was with the shin guard prior to the fracture in 16 cases (62%). Return to competitive soccer averaged 40 weeks for combined tibia and fibula fractures, 35 weeks for isolated tibia fractures, and 18 weeks for isolated fibula fractures. Injuries were associated with a high incidence of major complications (12 out of 31, 39%), especially in concurrent tibia and fibula fractures (8 out of 15, 50%). These findings suggest that lower leg fractures in soccer players are serious injuries, often necessitating a prolonged recovery time. In addition, this study questions the ability of shin guards to protect against fractures.

Concussion incidence in elite college soccer players.

A unique aspect of soccer is the use of the head for directing the ball. The potential for resultant head injuries has been the focus of discussions worldwide. Prior work has attributed neuropsychologic deficits to the cumulative effects of heading, without evaluating concussion rates in soccer players. We prospectively studied the seven men's and eight women's varsity soccer teams in the Atlantic Coast Conference during two seasons to document concussion incidence. The 29 concussions diagnosed over the 2 years in 26 athletes, 17 (59%) concussions in men and 12 (41%) in women, resulted from contact with an opponent's head (8, 28%), elbow (4, 14%), knee (1, 3%), or foot (1, 3%); the ball (7, 24%); the ground (3, 10%); concrete sidelines (1, 3%); goalpost (1, 3%); or a combination of objects (3, 10%). Twenty concussions (69%) occurred in games; none resulted from intentional heading of the ball. The basic incidence was 0.96 concussions per team per season. The overall incidence was 0.6 per 1000 athlete-exposures for men, and 0.4 per 1000 athlete-exposures for women. By concussion grade, there were 21 (72%) grade 1, 8 (28%) grade 2, and no grade 3 concussions. These findings suggest that concussions are more common in soccer than anticipated and that acute head injuries may have potential for long-term neuropsychologic changes.

Leg injuries and shin guards.

Soccer players have a high risk of soft-tissue injuries to their legs. Fractures of the tibia and fibula represent a serious potential injury; despite this, the incidence of these fractures in soccer players is unknown. Shin guards have become the only protective devices that are required by international and collegiate soccer associations. Nonetheless, the protective ability of shin guards has only been studied to a limited extent. Shin guards are assumed to be most effective at reducing leg abrasions and contusions. The role of shin guards in protecting against fractures has yet to be determined. Further analysis of the clinical effectiveness and biomechanical properties of shin guards is necessary to reduce the rate of leg fractures.