Behçet disease, new insights in disease associations and manifestations: a next-generation sequencing study.

Behçet disease is a multi-system disease associated with human leukocyte antigen (HLA) class I polymorphism. High-resolution next-generation sequencing (NGS) with haplotype analysis has not been performed previously for this disease. Sixty Egyptian patients diagnosed according to the International Study Group (ISG) criteria for Behçet disease and 160 healthy geographic and ethnic-matched controls were genotyped for HLA class I loci (HLA-A, B, C). For HLA class II loci (DRB1, DRB3/4/5, DQA1, DQB1, DPA1, DPB1), 40 control samples were genotyped. High-resolution HLA genotyping was performed using NGS and the results were analyzed. Clinical manifestations were oral ulcers (100%), genital ulcers (100%), eye (55%) and neurological (28%) and vascular involvement (35%). HLA-B*51:08 [odds ratio (OR) = 19·75, 95% confidence interval (CI) = 6·5-79; P < 0·0001], HLA-B*15:03 (OR = 12·15, 95% CI = 3·7-50·7; P < 0·0001), HLA-C*16:02 (OR = 6·53, 95% CI = 3-14; P < 0·0001), HLA-A*68:02 (OR = 3·14, 95% CI = 1·1-8·9; P < 0·01) were found to be associated with Behçet disease, as were HLA-DRB1*13:01 and HLA-DQB1*06:03 (OR = 3·39, 95% CI = 0·9-18·9; P = 0·04 for both). By contrast, HLA-A*03:01 (OR = 0·13, 95% CI = 0-0·8; P = 0·01) and HLA-DPB1*17:01 were found to be protective (OR = 0·27, 95% CI = 0·06-1·03; P = 0·02). We identified strong linkage disequilibrium between HLA-B*51:08 and C*16:02 and A*02:01 in a haplotype associated with Behçet disease. HLA-B*51:08 was significantly associated with legal blindness (OR = 2·98, 95% CI = 1·06-8·3; P = 0·01). In Egyptian Behçet patients, HLA-B*51:08 is the most common susceptibility allele and holds poor prognosis for eye involvement.

Presentation outside office hours does not negatively influence treatment times for reperfusion therapy for acute ischemic stroke.

BACKGROUND: Treatment outside office hours has been associated with increased workflow times for intravenous thrombolysis (IVT) in acute ischemic stroke (AIS). Limited data suggest that this "off-hours effect" also exists for endovascular treatment (EVT). We investigated this phenomenon in a well-organized acute stroke care region in the Netherlands. METHODS: Retrospective, observational cohort study of consecutive patients with AIS who received reperfusion therapy in the Greater Amsterdam Area, consisting of 14 primary stroke centers and 1 comprehensive stroke center (IVT: 2009-2015, EVT: 2014-2017). Office hours were defined as presentation during weekdays between 8 AM and 5 PM, excluding National Festive days. Primary outcome was door-to-treatment time (door-to-needle [DNT] for IVT, door-to-groin [DGT] for EVT). For DGT, we used the door time of the first hospital. Other outcomes were in-hospital mortality, modified Rankin Scale (mRS) score at 90 days and symptomatic intracranial hemorrhage (sICH). We performed multivariable linear and logistic regression analyses and used multiple imputation to account for missing values. RESULTS: In total, 59% (2450/4161) and 61% (239/395) of patients treated with IVT and EVT, respectively, presented outside office hours. Median DNT was minimally longer outside office hours (32 vs. 30 min, p = 0.024, adjusted difference 2.5 min, 95% CI 0.7-4.2). Presentation outside office hours was not associated with a longer DGT (median 130 min for both groups, adjusted difference 7.0 min, 95% CI - 4.2 to 18.1). Clinical outcome and sICH rate also did not differ. CONCLUSION: Presentation outside office hours did not lead to clinically relevant treatment delays for reperfusion therapy in patients with AIS.

Mitochondrial dysfunction increases pro-inflammatory cytokine production and impairs repair and corticosteroid responsiveness in lung epithelium.

COPD is characterized by chronic lung inflammation and irreversible lung tissue damage. Inhaled noxious gases, including cigarette smoke, are the major risk factor for COPD. Inhaled smoke first encounters the epithelial lining of the lungs, causing oxidative stress and mitochondrial dysfunction. We investigated whether a mitochondrial defect may contribute to increased lung epithelial pro-inflammatory responses, impaired epithelial repair and reduced corticosteroid sensitivity as observed in COPD. We used wild-type alveolar epithelial cells A549 and mitochondrial DNA-depleted A549 cells (A549 Rho-0) and studied pro-inflammatory responses using (multiplex) ELISA as well as epithelial barrier function and repair (real-time impedance measurements), in the presence and absence of the inhaled corticosteroid budesonide. We observed that A549 Rho-0 cells secrete higher levels of pro-inflammatory cytokines than wild-type A549 cells and display impaired repair upon wounding. Budesonide strongly suppressed the production of neutrophil attractant CXCL8, and promoted epithelial integrity in A549 wild-type cells, while A549 Rho-0 cells displayed reduced corticosteroid sensitivity compared to wild-type cells. The reduced corticosteroid responsiveness may be mediated by glycolytic reprogramming, specifically glycolysis-associated PI3K signaling, as PI3K inhibitor LY294002 restored the sensitivity of CXCL8 secretion to corticosteroids in A549 Rho-0 cells. In conclusion, mitochondrial defects may lead to increased lung epithelial pro-inflammatory responses, reduced epithelial repair and reduced corticosteroid responsiveness in lung epithelium, thus potentially contributing to the pathogenesis of COPD.

Should internal mammary chain (IMC) sentinel node biopsy be performed? Outcome in 90 consecutive non-biopsied patients with a positive IMC scintigraphy.

INTRODUCTION: Although the status of the regional lymph nodes is an important determinant of prognosis in breast cancer, harvesting sentinel nodes (SN) detected in the internal mammary chain (IMC) is still controversial. AIMS: To determine in how many patients a positive IMC-SN might change the systemic or locoregional adjuvant therapy, with a possible benefit in outcome. PATIENTS AND METHODS: During 6 1/2 years data of T1-2 breast cancer patients, having an SN procedure, were prospectively collected. Our policy was not to explore the IMC even if it was the only localization of an SN. RESULTS: In 86 of 571 patients lymphoscintigraphy showed an IMC-SN. In 64 of these, the axillary SN was negative and only 25 of these patients did not have an indication for adjuvant systemic treatment based on their tumor characteristics. In the literature, IMC metastases are found in 0-10% of axillary negative patients. Routine IMC-SN biopsies would have resulted in an indication for adjuvant systemic therapy in 2-3 of our patients. Four parasternal recurrences were found during a median follow-up of 51 months. CONCLUSIONS: Harvesting IMC-SNs is a procedure of which only a limited number of patients have therapeutical benefit. Even with a thorough selection of patients, the extra morbidity of the procedure should be weighed against the potential benefit for the patient.

The influence of age on the pharmacokinetics of the antiepileptic agent oxcarbazepine.

The disposition of oxcarbazepine was studied in 12 young and 12 elderly healthy male and 12 young and 12 elderly healthy female volunteers, with emphasis on the influence of age. Oxcarbazepine was administered as a single dose of either 300 mg (men) or 600 mg (women), followed by multiple-dose (300 mg) administration twice a day for 7 days (men) or 6 days (women). Semilogarithmic plasma concentration-time curves showed an increasing decline at decreasing concentrations. Accumulation of the pharmacologically active metabolite monohydroxycarbamazepine was found to be more than one would anticipate on the basis of linear and unchanged pharmacokinetics. Saturation did not seem to occur at the level of renal excretion. No apparent differences between male and female volunteers were observed. A significant higher maximum concentration, higher area under the curve parameters, and a lower elimination rate constant were observed in the elderly. These observations are in line with a smaller renal clearance of monohydroxycarbamazepine in the elderly group. In a clinical situation, these age-related differences are not likely to have important implications. In general, treatment with oxcarbazepine was well tolerated.

Neurolymphomatosis of the median nerve.

A patient with a non-Hodgkin's lymphoma had a painful axonal neuropathy of the median nerve due to lymphomatous infiltration. The median nerve lesion was the only site of tumor recurrence for 5 months and could be diagnosed with MRI. The median neuropathy responded to chemotherapy.

99mTc-SPECT in the diagnosis of skull base metastasis.

OBJECTIVE: To evaluate the role of bone SPECT in diagnosing metastasis to the skull base in cancer patients. BACKGROUND: Skull base metastasis often causes characteristic clinical syndromes. Antitumor treatment ameliorates or stabilizes symptoms in most patients. Diagnosis may be difficult when neuroimaging studies are negative (as occurs in about one-quarter of patients). Case reports have suggested a role for bone SPECT in these patients. METHODS: We reviewed the charts of all patients (1993-1996) at our institution who had skull base SPECT and at least one neuroimaging study (CT or MRI) for clinically suspected metastasis to the base of the skull. Bone SPECT, CT, and MRI were blindly re-evaluated. RESULTS: We studied 56 patients, of whom 36 had skull base metastasis. Twenty patients had other causes of the clinical syndrome, including leptomeningeal and posterior fossa metastasis, or benign causes. In 29 of 36 patients (81%) with skull base metastasis, CT or MRI clearly demonstrated the lesion. Bone SPECT identified a hot spot in the appropriate region of the skull base in 28 of 36 patients (78%). All seven patients with negative CT or MRI had positive SPECT and four of these had a response to anti-tumor treatment. CONCLUSIONS: SPECT of the skull base can demonstrate lesions not identifiable by CT or MRI. In cancer patients suspected of having skull base metastasis, we recommend SPECT of the skull base when CT or MRI studies are negative.

Effect of an evening dose of regular and effervescent formulations of ranitidine or cimetidine on intragastric pH in healthy volunteers.

AIMS: To compare the effects on intragastric acidity of a single evening dose of either standard or effervescent formulations of ranitidine (300 mg) or cimetidine (800 mg). METHODS: Twelve healthy subjects were studied, using a four-period randomized cross-over design and an ambulatory intragastric pH monitoring technique. The subjects received a standard evening meal at 17.00 hours and one of the H2-receptor antagonist formulations was given at 23.00 hours. RESULTS: Both effervescent formulations caused a transient rapid increase in intragastric pH, reaching a maximum at about 3 min after ingestion. After both effervescent formulations a significantly higher pH was measured during the first 45 min after ingestion (P < 0.05), compared to the regular formulations. The onset of action of the H2-receptor antagonists was similar for both formulations of ranitidine and the effervescent cimetidine, but tended to be slower for the regular cimetidine (P = 0.06). Nocturnal intragastric pH was significantly increased by all four formulations, but more effectively so by the two ranitidine formulations. The duration of action (taken as time with pH > 4) of both ranitidine formulations was longer than that of both cimetidine formulations (P < 0.002). CONCLUSIONS: A single evening dose of 300 mg ranitidine produces a stronger decrease of nocturnal gastric acid secretion than 800 mg cimetidine. The effervescent formulations of both drugs offer the advantage of a rapid decrease (within minutes) of intragastric acidity, with preservation of the sustained systemic effect.

The diagnostic accuracy of magnetic resonance imaging and cerebrospinal fluid cytology in leptomeningeal metastasis.

Diagnostic decision making in the case of patients suspected of having leptomeningeal metastasis (LM) can be very difficult. The results of cerebrospinal fluid (CSF) cytology can be repeatedly negative, and the predictive value of gadolinium-enhanced magnetic resonance imaging (MRI) is not well known. We report the results of CSF cytology and Gd MRI in 61 patients with known cancer, suspected of having LM. We combined our data with those from a similar study and calculated the sensitivity and specificity of CSF and Gd MRI, in the absence of a "gold standard diagnosis." CSF cytology was positive for LM in 35 patients and MRI in 38. With CSF cytology sensitivity 75% and specificity 100%, with Gd MRI sensitivity was 76% but specificity only 77%. We conclude that Gd MRI provides strong support in the diagnosis of LM in patients with cancer who have negative results on CSF cytology.

Spinal epidural abscess complicating chronic epidural analgesia in 11 cancer patients: clinical findings and magnetic resonance imaging.

We reviewed the records of all patients who had received an epidural catheter for management of chronic cancer pain in a 3-year period (1993-1996). Patients with nervous system infections were identified, and pertinent clinical, radiological (magnetic resonance imaging), and bacteriological data were analyzed. We identified 11 patients who developed spinal epidural abscess (SEA). All of these had back pain; radicular signs occurred in seven patients and spinal cord compression in two patients. Magnetic resonance imaging revealed SEA in all 11 patients. SEA was iso- to hypointense on T1-weighted images and hyperintense on T2-weighted images relative to spinal cord. After gadolinium administration seven lesions showed characteristic rim enhancement while three showed minimal enhancement. No signs of diskitis or osteomyelitis were present, and the abscess was always localized to the posterior epidural space. Cultures were positive in all cases and revealed Staphylococcus epidermidis in eight and S. aureus in three. All patients were treated with intravenous antibiotics, and four had an additional decompressive laminectomy. Two patients died within 1 week of diagnosis from overwhelming septicemia despite apparently adequate antibiotic treatment. Within 4 weeks after diagnosis of SEA two patients died from widely metastatic disease, although infection may have contributed. One patient developed septicemia while receiving appropriate antibiotics and underwent emergency laminectomy. The neurological deficits recovered in all patients who survived the acute infectious episode. We conclude that patients with chronic epidural catheters for cancer pain require prompt neurological evaluation and magnetic resonance imaging when SEA is suspected. Early evaluation and treatment may lead to full recovery.

Negative sural nerve biopsy in neurolymphomatosis.

Patients with non-Hodgkin's lymphoma occasionally develop widespread invasion of peripheral nerves by tumor cells or neurolymphomatosis (NL). Clinically this usually results in asymmetrical, progressive, and painful polyneuropathy. Diagnosis rests on the identification of tumor cells in peripheral nerves. To avoid false-negative biopsy findings in patients with malignant lymphomatous infiltration of peripheral nerves it has been recommended to biopsy clinically involved nerves. We present two patients with histologically confirmed NL in whom sural the nerve biopsy finding was negative despite clinical and neurophysiological evidence of involvement of the sural nerve a. The clinical features of NL are reviewed. Some patients with neurolyphomatosis have only focal or proximal involvement of nerves, requiring the biopsy of an affected part of these nerves. Magnetic resonance imaging may be useful in identifying affected nerves.

Concentration changes in surface Ca, P, F, Zn, Fe, and Sr during white spot formation.

The purpose of this study was to investigate the mechanism of surface layer formation in artificial white spots. Two nondestructive nuclear micro-analysis techniques were employed to analyze windows of human dental enamel at progressive stages of an artificial lesion. Fluoride concentrations were measured to a depth of 9 micron using resonant reaction profiling. Surface concentrations of Ca, P, Zn, Fe, and Sr were measured using proton-induced X-ray emission. The enamel windows were analyzed longitudinally after zero, four, and 16 hours' exposure to lactic acid gels of pH 3.7 and pH 4.5. During the first four hr of lesion formation, nonfluoridated apatite was preferentially removed from the surface with up to a 40% loss of Ca and P, depending on the gel pH. The next 12 hr saw a build-up of Ca and P in the surface to values approaching those of healthy enamel. In comparison, F and Sr concentrations changed very little, Fe tended to show a small increase at the end of each demineralization period, and Zn was less readily lost and gained than was Ca. These results and others suggest that the white spot surface layer phenomenon is a combined demineralization/remineralization process aided by, but not requiring, the presence of endogenous fluoride.

Shallow fluorine depth profiles of cementum in periodontal disease--a pilot study.

A nuclear resonant reaction depth profiling technique was used to analyze elemental fluorine distribution in the first 20 microns of human dental cementum. A pilot sample of six periodontally-involved teeth indicated greater levels of fluorine in exposed cementum (0.9 leads to 2.4%) compared to cementum apical to the zone of epithelial attachment (0.4 leads to 1.1%). Furthermore, the exposed cementum appeared to have fluorine levels within the surface 5 microns comparable to the hypermineralized layer previously reported by x-ray diffraction and microprobe techniques. The nuclear resonant reaction is a non-destructive technique which yields useful information of surface elemental distribution as a function of depth, and may be regarded as a potential means of analyzing changes in the inorganic constitution of cementum during various physico-chemical pre-treatments.

Magnetic resonance imaging of the coronary arteries: imaging planes and resulting anatomy in two-dimensional imaging.

Magnetic-resonance imaging techniques use different imaging planes than does conventional coronary angiography to acquire longer segments of a coronary artery in a single tomographic slice. At first sight, these planes appear rather puzzling, because the coronary arteries are displayed in unfamiliar orientations. In this article we will review the existing methodology for obtaining the orientations for the proximal coronary arteries and describe the associated anatomical landmarks that can be seen. Additional orientations for the middle segment of the circumflex and distal right coronary artery are introduced. These orientations are used both in various acquisition techniques and for evaluation of three-dimensional data when using multiplanar reformatting.

The effect of anisotropy on the potential distribution in biological tissue and its impact on nerve excitation simulations.

We present a finite difference solution of the potential distribution associated with electrical current stimulation in an anisotropic in-homogeneous tissue environment and compare it to the isotropic case. The results demonstrate that there can be significant errors associated with the assumption of isotropic tissue properties in calculating the potential distribution along an axon in nerve excitation simulations. These errors can have a significant impact on predicted nerve fiber recruitment patterns when evaluating the efficacy of specific surface or intramuscular stimulus electrode configurations. The results of this study also suggest when a more comprehensive tissue model should be implemented in an electrode design study. Simulation results indicate that the isotropy assumption is worst under bipolar electrode stimulation as opposed to monopolar stimulation and that the bipolar error increases as the distance between electrodes decreases. In light of these results, it is concluded that in order to avoid large errors in the calculated potential distribution along an axon, the isotropy assumption should only be used when the transverse depth from the electrode to the nerve is relatively small.

Bacteriophages and Clostridium spores as indicator organisms for removal of pathogens by passage through saturated dune sand.

In a field study on the efficiency of dune recharge for drinking water production, bacteriophage MS2 was shown to be removed 8 log(10) by passage through the dune sand. The question of whether pathogenic viruses would be removed as much as MS2 was studied by comparing complete breakthrough curves of MS2 with those of the human viruses Coxsackievirus B4 (CB4) and Poliovirus 1 (PV1) in laboratory columns. The columns were designed to closely simulate the field conditions: same sand, water, porewater velocity and temperature. Employing a two-site kinetic model to simulate breakthrough curves, attachment/detachment to two types of kinetic sites as well as inactivation of free and attached viruses were evaluated. It was found that attachment to only one of the sites is of significance for determining overall removal. At field scale, removal of the less negatively charged PV1 was extrapolated to be about 30 times greater than that of MS2, but removal of CB4 would be only as much as that of MS2. Also, removal of spores of Clostridium perfringens D10, a potential surrogate for Cryptosporidium oocysts, was studied. The attachment rate coefficient of the spores was 7.5 times greater than that of MS2. However, this does not imply that the removal of the spores is 7.5 times greater than that of MS2. Due to negligible inactivation in combination with detachment of previously attached spores, the actual removal rate of the spores depends on the duration of contamination and eventually all spores will break through. Provided no irreversible attachment or physical straining occurs, this may also be the case for other persistent microorganisms, like oocysts of Cryptosporidium.

Motor unit populations in healthy and diseased muscles.

The numbers of functioning motor units can be estimated in proximal and distal muscles of human limbs by an electrophysiological technique in which the mean sizes of the motor unit potentials are compared with the maximum M-waves of the same muscles. Although manual methods of estimation have been used successfully in the past, the introduction of automated techniques has brought considerable advantages, including greater objectivity and reduced contamination of the results by "alternation." In healthy subjects, the intrinsic muscles of the hand have approximately 100 motor units each, and the biceps brachii muscle has only slightly more. With advancing age, there is a loss of motor units, which appears to be more pronounced in distal muscles. The motor unit estimating methodology has been found to be of value in the diagnosis and assessment of patients suspected of having muscle denervation. In amyotrophic lateral sclerosis, the mean rate of motor unit loss is swift, whereas in late-onset cases of spinal muscular atrophy, the reduction in the motor unit population does not appear to progress. In only the most rapidly deteriorating cases of post-polio syndrome is it possible to demonstrate further loss of motor units. In all of these denervating disorders, and in peripheral neuropathies, the importance of collateral reinnervation as a compensatory mechanism is emphasized.

The effect of stimulus current pulse width on nerve fiber size recruitment patterns.

There have been theoretical studies presented that postulate a change in the stimulus current amplitude required to recruit nerve fibers with different stimulus current pulse widths. Based on these theoretical predictions, it has been suggested that the stimulus pulse width parameter may be used to selectively recruit fibers of different sizes and that this selectivity should increase with increasing distance from the stimulus electrode. In this paper, a simulation study of the recruitment patterns of a population of motor nerve fibers with a histologically accurate fiber diameter distribution is presented. Nerve fiber excitation simulations coupled with a time varying field simulation suggest that, for surface stimulation, there is only a marginal selectivity achievable in the average nerve fiber diameter that is recruited across the range of commonly used stimulus pulse widths but this selectivity also increases with increased electrode distance. Experimental evidence consisting of estimates of nerve fiber diameter based on motor unit latency studies is also presented that is consistent with the predictions made by the electromagnetic field and nerve fiber excitation simulations.

Column experiments to study nonlinear removal of bacteriophages by passage through saturated dune sand.

In a recent field study on dune recharge, bacteriophages MS2 and PRD1 were found to be removed 3 log10 over the first 2.4 m and only 5 log10 over the next 27 m. To understand the causes of this nonlinear removal, column experiments were carried out under conditions similar to the field: same recharge water, temperature (5 +/- 3 degrees C) and pore water velocity (1.5 m day(-1)). Soil samples were taken along a streamline between the recharge canal and the first monitoring well. Bacteriophage phiX174 was included for comparison. The high initial removal in the field was found not to be due to heterogeneity of phage suspensions but to soil heterogeneity. Phage removal rates correlated strongly positively with soil organic carbon content, and relatively strongly positively with silt content and the presence of ferric oxyhydroxides. Soil organic carbon content, silt content and the presence of ferric oxyhydroxides were found to decrease exponentially with travel distance. Removal rates of phiX174 were found to be 3-10 times higher than those of MS2 and PRD1 due to the lower electrostatic repulsion that the less negatively charged phiX174 experiences. It is suggested that the high initial removal in the field is due to the presence of favorable sites for attachment formed by ferric oxyhydroxides that decrease exponentially with travel distance. Similar removal rates may be found at both laboratory and field scale. However, due to local variations at field scale detailed knowledge on soil heterogeneity may be needed to enable a reliable prediction of removal.

Minute-by-minute differences in co-activation during treadmill walking in cerebral palsy.

The primary purpose of this study was to determine, in children and adolescents with mild spastic cerebra palsy (CP); 1) minute-by-minute differences in lower limb antagonist muscle co-activation and stride length (SL) during treadmill walking following 12-15 minutes of treadmill walking practice, and 2) if the minute-by-minute pattern of co-activation is affected by site (thigh or lower leg) and lower limb dominance. A secondary purpose was to determine if overall there is a difference in co-activation between the dominant and non-dominant lower limbs. Eight independently ambulatory children and adolescents with mild spastic CP (9.2-15.7 yr) participated in the study. Minute-by-minute lower limb antagonist muscle co-activation and SL were measured during a 3-minute treadmill walk at 90% of individually determined fastest treadmill walking speed. Non-dominant thigh (quadriceps, hamstring muscles) co-activation decreased between minute 1 and a) minute 2 (6%), b) minute 3 (7.2%). Co-activation for the dominant lower leg (tibialis anterior, triceps surae muscles) decreased between minute 1 and minute 3 (11.3%). Non-dominant thigh co-activation was on average 27.3% higher than for the dominant thigh. Thigh co-activation was on average 27.7% higher than for the lower leg, independent of dominance or time. SL increased between minute 1 and minute 3 by 2.1%. Twelve to 15 minutes of treadmill walking practice may be sufficient time to obtain stable co-activation and SL values by minute 2 of a fast treadmill walk. Dominance and site affect the magnitude of co-activation.