Effects of different surgical modalities on the clinical outcome of patients with moyamoya disease: a prospective cohort study.
OBJECTIVE Bypass surgery is the most common treatment for moyamoya disease (MMD), but there is controversy over which surgical modality is best. The objective of this study was to evaluate the clinical outcome of patients with MMD after undergoing different surgical modalities. METHODS A series of 696 consecutive MMD patients treated between June 2009 and May 2015 were screened in this prospective cohort study. Patients who did not undergo revascularization surgeries and those who underwent different surgical modalities in bilateral hemispheres were excluded. Finally, 529 patients who were observed for at least 12 months were included: 438 patients underwent unilateral surgery, and 91 patients underwent bilateral surgery. Of these, 241 patients underwent direct bypass (DB); 81, a combined bypass (CB); and 207, an indirect bypass (IB). Three clinical outcomes were evaluated and compared between surgical groups: recurrent stroke events, modified Rankin Scale (mRS) scores, and change in the main symptoms. RESULTS The mean follow-up period was 40 months. During the follow-up period, recurrent stroke was observed in 43 patients, including 15 patients with hemorrhage, 26 patients with ischemia (transient ischemic attack in 19 patients and infarction in 7 patients), and 2 patients with both hemorrhage and cerebral infarction. Kaplan-Meier analysis showed that patients who underwent a CB or DB had a longer ischemia-free time than those who underwent IB (p = 0.013); however, there was no significant difference in the hemorrhage-free time between the different surgical modalities (p = 0.534). A good neurological status (mRS score ≤ 2) was achieved in 495 patients (93.6%) and was significantly achieved by more children (98.2%) than adults (92.3%; p = 0.022). Surgical modalities were not significantly associated with outcome neurological status (p = 0.860). Moreover, improvement in symptoms was observed in 449 patients (84.9%) and was also significantly more common in children (93.0%) than in adults (82.7%; p = 0.006). No significant difference was observed between the different surgical modalities (p = 0.548). CONCLUSIONS CB and DB are more effective at preventing recurrent ischemic strokes than IB. However, there is no evidence that these 3 surgical modalities demonstrate significant differences in preventing recurrent hemorrhage.


Muscle oxygenation and time to task failure of submaximal holding and pulling isometric muscle actions and influence of intermittent voluntary muscle twitches.
Isometric muscle actions can be performed either by initiating the action, e.g., pulling on an immovable resistance (PIMA), or by reacting to an external load, e.g., holding a weight (HIMA). In the present study, it was mainly examined if these modalities could be differentiated by oxygenation variables as well as by time to task failure (TTF). Furthermore, it was analyzed if variables are changed by intermittent voluntary muscle twitches during weight holding (Twitch). It was assumed that twitches during a weight holding task change the character of the isometric muscle action from reacting (≙ HIMA) to acting (≙ PIMA). Twelve subjects (two drop outs) randomly performed two tasks (HIMA vs. PIMA or HIMA vs. Twitch, n = 5 each) with the elbow flexors at 60% of maximal torque maintained until muscle failure with each arm. Local capillary venous oxygen saturation (SvO2) and relative hemoglobin amount (rHb) were measured by light spectrometry. Within subjects, no significant differences were found between tasks regarding the behavior of SvO2 and rHb, the slope and extent of deoxygenation (max. SvO2 decrease), SvO2 level at global rHb minimum, and time to SvO2 steady states. The TTF was significantly longer during Twitch and PIMA (incl. Twitch) compared to HIMA (p = 0.043 and 0.047, respectively). There was no substantial correlation between TTF and maximal deoxygenation independently of the task (r = - 0.13). HIMA and PIMA seem to have a similar microvascular oxygen and blood supply. The supply might be sufficient, which is expressed by homeostatic steady states of SvO2 in all trials and increases in rHb in most of the trials. Intermittent voluntary muscle twitches might not serve as a further support but extend the TTF. A changed neuromuscular control is discussed as possible explanation.


Synthesis and structure-activity relationships of potent new angiotensin converting enzyme inhibitors containing saturated bicyclic amino acids.
The synthesis of a series of novel, potent angiotensin converting enzyme (ACE) inhibitors containing saturated bicyclic amino acids in place of proline is described. Octahydroindole-2-carboxylic acid, octahydroisoindole-1-carboxylic acid, and octahydro-3-oxoisoindole-1-carboxylic acid can replace proline in both sulfhydryl and non-sulfhydryl ACE inhibitors to give compounds equipotent to captopril and enalapril both in vitro and in vivo. Structure-activity relationships are discussed. Compound 11a (CI-907, indolapril) has advanced to clinical evaluation.


Molecular actions of Escherichia coli MutT for control of spontaneous mutagenesis.
MutT protein of Escherichia coli hydrolyzes oxidized guanine nucleotides, 8-oxo-dGTP and 8-oxoGTP, to the corresponding monophosphates, thereby preventing misincorporation of 8-oxoguanine into DNA and RNA, respectively. Although the biological significance of the MutT has been established, how MutT protein actually works in vivo remains to be elucidated. The current study shows the molecular behavior of the MutT protein in vivo and in vitro with special reference to control of spontaneous mutagenesis. A single E. coli cell carries about 70-75 molecules of the MutT protein and that this number does not change even when the cells were cultured in anaerobic and hyper-oxidative conditions. Conditional gene silencing analyses revealed that about a half number of MutT molecules are needed for keeping the spontaneous mutation frequency at the normal level. The MutT functions are not needed under anaerobic condition, yet the level of the MutT protein in cell is kept constant, probably for preparing for sudden changes of oxygen pressure. There is a possibility that MutT functions in close association with other proteins, and evidence is presented that MutT protein can interact with some proteins in vivo.


Veno-occlusive disease of the liver following high-dose chemotherapy and autologous bone marrow transplantation in children with solid tumors: incidence, clinical course and outcome.
Two hundred and thirty-six consecutive courses of high-dose chemotherapy with autologous bone marrow transplantation in children with solid tumors were reviewed in order to assess the incidence, clinical presentation and outcome of veno-occlusive disease (VOD) of the liver. Patients conditioned with total body irradiation were excluded from this study. Eleven patients (4.6%) met the diagnostic criteria for VOD. The clinical course included sudden weight gain, jaundice, hepatomegaly and ascites. Renal dysfunction and refractoriness to platelet transfusions occurred in the most severe forms. Seven patients recovered within 7-29 days of onset and four patients died, all with renal failure and fluid overload. The time of onset appeared to determine two patterns of outcome: mild forms with early onset (before day 11) and more severe forms with onset after day 17. Analysis of pretransplant factors revealed no significant association with an increased risk of VOD. However, all the patients with severe VOD had received a conditioning regimen containing cyclophosphamide which might be involved in the pathogenesis of VOD.


Derivation of leukemic plasmacytoid dendritic cells coexpressing a progenitor cell surface antigen, CD117, without interferon-alpha production.
CD4+CD56+ lineage negative malignancy has recently been considered as plasmacytoid dendritic cell (PDC) leukemia/lymphoma. We investigated immunophenotypic and functional characterizations of PDC leukemic cells in one case. Lineage markers were all negative except partially positive CD11c and positive CD117, indicating malignant cells were leukemic PDCs coexpressing myeloid and progenitor cell surface antigens. Leukemic PDCs cultured with IL-3 increased in size and expression of CD11c, CD40 and HLA-DR, although the cells cultured with IL-2 or GM-CSF showed little proliferation. Furthermore, CD40 ligation after IL-3 stimulation yielded morphological changes such as expression of dendritic process. These findings showed that malignant cells were consistent with leukemic PDCs. However, secretion of interferon-alpha was not detected in leukemic PDCs with the stimulation of CpG ODN or inactivated herpes simplex virus-1.


The electrophysiological effects of ouabain on sinus node and atrium in man.
Electrophysiological studies were performed in 16 patients before and 30 min after intravenous administration of ouabain (0.1 mg/kg). P-A interval (mean+/-SEM) was 40+/-2.1 ms before and 44+/- 1.5 ms after ouabain (P less than 0.001). Atrial effective and functional refractory periods (ERP and FRP) were measured in all patients during sinus rhythm and during driving at equivalent paced rates in 12 patients. The mean atrial ERP and FRP during sinus rhythm were, respectively, 244+/-10.5 and 307+/-11.0 ms before and 253+/-9.7 and 318+/-11.4 ms after infusion of ouabain (NS). Mean atrial ERP and FRP during driving were, respectively, 231+/-15.3 and 264+/-14.9 ms before and 266+/-18.6 and 296+/-19.7 ms after ouabain (P less than 0.01 and P less than 0.01). Mean sinus cycle length and sinus recovery times were, respectively, 887+/-31.2 and 1,113+/-38.7 ms before and 905+/-38.2 and 1,008+/-30.7 ms after infusion of ouabain (NS and P less than 0.005). Calculated sinoatrial conduction times before and after ouabain were 90+/-6.8 and 110+/-8.5 ms, respectively (P less than 0.005). In summary, ouabain produced depression of intraatrial conduction as manifested by increase in P-A interval and atrial effective and functional refractory periods. Ouabain significantly increased calculated sinoatrial conduction time without significant effect on spontaneous sinus cycle length.


Effects of age on rat glutathione metabolism.
The authors hypothesized that rat plasma or tissue glutathione metabolism could change with age due to possible decreases in glutathione-related enzyme activities. To test this hypothesis, the authors measured plasma and tissue concentrations of glutathione and glutathione-related enzymes. Animals were 3 months, 12 months, or 24 months old at the time of experiments. Plasma glutathione was found to be significantly increased in both the 12-month-old and 24-month-old groups compared to the 3-month-old rats. Tissue enzyme measurements showed no significant differences between the groups in lung or liver glutathione peroxidase or glutathione S-transferase. gamma-Glutamyl transpeptidase activity was significantly decreased in kidney and lung with aging. Decreases in tissue gamma-glutamyl transpeptidase activity occur with age; this may contribute to increases in plasma glutathione concentrations.


Homozygous DNA variants in exon 9 of the LDL receptor gene in a Thai patient with primary hypercholesterolemia phenotype.
Mutation in low density lipoprotein (LDL) receptor gene causes an inherited primary hypercholesterolemia namely familial hypercholesterolemia (FH). In this study, 46 Thai patients with primary hypercholesterolemia were screened for mutations in exon 9 of the LDL receptor gene by polymerase chain reaction-restriction fragment length polymorphism (PCR - RFLP). The analysed fragment was 224 bp in length. According to the published cDNA sequence, exon 9 of the LDL receptor gene contains several hypermutable CpG dinucleotides. Three of these sites are Hpa II recognition sites. PCR product of exon 9 obtained from amplification of wild-type DNA sample would yield four fragments after Hpa II digestion. The expected sizes of these restriction fragments were 15, 30, 40 and 139 bp. All normocholesterolemic subjects (n = 33) showed normal RFLP. However, in one patient (72 year old female), abnormal RFLP from Hpa II digestion of the amplified exon 9 was observed, i.e., a fragment of 70 bp and another one smaller than 139 bp. Such RFLP reflects that exon 9 of both alleles of the LDL receptor gene in this patient lost one and gained one Hpa II site. It is interesting that this patient, eventhough harbouring two mutations on both alleles of the LDL receptor gene (presumably homozygous genotype of FH), apparently revealed lipid levels of heterozygous phenotype of FH without symptoms of coronary artery disease. It has yet to be proved whether these genetic variations are disease-related mutations or presumably common DNA polymorphisms.


Urothelial carcinoma of bladder in a pediatric patient case report of a rare disease.
Urothelial carcinoma is a very rare malignancy in the pediatric population. In spite of the extremely low amount of published cases, the incidence rate is between 0.4 and 0.1% before the age of 20. Insomuch that only less than 30 cases have been reported in the first decade. Those tumors were mostly solitary, non-invasive, with low-risk of progression and recurrence rate compared to adult-onset form. In this case report, we aim to discuss the diagnosis and treatment of a 10-year-old male patient with urothelial carcinoma of bladder who admitted to our clinic.


Enantiomeric assay of escitalopram S(+)-enantiomer and its "in-process impurities" using two different techniques.
The enantiomeric purity of escitalopram oxalate ESC and its "in-process impurities," namely, ESC-N-oxide, ESC-citadiol, and R(-)-enantiomer were studied in drug substance and products using high-performance liquid chromatography (HPLC)-UV (Method I), synchronous fluorescence spectroscopy (SFS) (Method IIA), and first derivative SFS (Method IIB). Method I describes as an isocratic HPLC-UV for the direct resolution and determination of enantiomeric purity of ESC and its "in-process impurities." The proposed method involved the use of αl -acid glycoprotein (AGP) chiral stationary phase. The regression plots revealed good linear relationships of concentration range of 0.25 to 100 and 0.25 to 10 μg mL-1 for ESC and its impurities. The limits of detection and quantifications for ESC were 0.075 and 0.235 μg mL-1 , respectively. Method II involves the significant enhancement of the fluorescence intensities of ESC and its impurities through inclusion complexes formation with hydroxyl propyl-β-cyclodextrin as a chiral selector in Micliavain buffer. Method IIA describes SFS technique for assay of ESC at 225 nm in presence of its impurities: R(-)-enantiomer, citadiol, and N-oxide at ∆λ of 100 nm. This method was extended to (Method IIB) to apply first derivative SFS for the simultaneous determination of ESC at 236 nm and its impurities: the R(-)-enantiomer, citadiol, and N-oxide at 308, 275, and 280 nm, respectively. Linearity ranges were found to be 0.01 to 1.0 μg mL-1 for ESC and its impurities with lower detection and quantification limits of 0.033/0.011 and 0.038/0.013 μg mL-1 for SFS and first derivative synchronous fluorescence spectra (FDSFS), respectively. The methods were used to investigate the enantiomeric purity of escitalopram.


The ins and outs of venous access: part I.
Selection of the proper venous access device is important to maximize patient benefit and minimize patient discomfort, morbidity, mortality, and cost. The decision of which device to use is based on whether or not the patient requires central venous access and whether the need is short-term (<6 to 8 weeks) or long-term. Short-term venous access devices include short peripheral IV catheters, midline catheters, peripherally inserted central catheters (PICC), and central venous catheters (CVC). This article reviews each of these short-term devices and their indications, contraindications, advantages, and disadvantages. Part 1 covers Venous Anatomy and Short-Term Venous Access; Part 2, to be published in the June issue, covers Long-Term Venous Access.


Partition functions and metropolis-type evolution rules for surface growth models with constraints.
We study dynamical scaling properties of the surface growth model with the Metropolis-type evolution rule from a partition function Z= sum ([h(r)])II (h(max))(h=h(min))1/2(1+z(n(h))), where z is a fugacity-like quantity and n(h) is the number of sites with height h in a surface configuration [h(r)]. The partition function describes a 2-particle correlated growth model when z=-1 and a self-flattening growth model when z=0. For one-dimensional equilibrium surfaces, the scaling properties for z>or=-1 except z=1 are all one phase with roughness exponent alpha=1/3 and growth exponent beta approximately equal 0.22. For the growing (eroding) surfaces, there exists a phase transition at z=0 from the grooved phase (alpha=1) for -1<or=z<0 to the ordinary Kardar-Parisi-Zhang phase (alpha=1/2) for z>0.


Synthesis and application of a ternary composite of clay, saw-dust and peanut husks in heavy metal adsorption.
The adsorption of a multi-component system of ferrous, chromium, copper, nickel and lead on single, binary and ternary composites was studied. The aim of the study was to investigate whether a ternary composite of clay, peanut husks (PH) and saw-dust (SD) exhibited a higher adsorption capacity than that of a binary system of clay and SD as well as a single component adsorbent of PH alone. The materials were used in their raw state without any chemical modifications. This was done to retain the cost effective aspect of the naturally occurring adsorbents. The adsorption capacities of the ternary composite for the heavy metals Fe2+, Cr3+, Cu2+, Ni2+ and Pb2+ were 41.7 mg/g, 40.0 mg/g, 25.5 mg/g, 41.5 mg/g and 39.0 mg/g, respectively. It was found that the ternary composite exhibited excellent and enhanced adsorption capacity compared with both a binary and single adsorbent for the heavy metals Fe2+, Ni2+ and Cr3+. Characterization of the ternary composites was done using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and thermogravimetric analysis (TGA). Kinetic models and adsorption isotherms were also studied. The pseudo second order kinetic model and the Langmuir adsorption isotherm best described the adsorption mechanisms for the ternary composite towards each of the heavy metal ions.


Hydrodynamic mobility of an optically trapped colloidal particle near fluid-fluid interfaces.
Using optical tweezers, we measure the anisotropic hydrodynamic mobility of a colloidal particle by tracking its thermal motion in an optical trap located near fluid-fluid interfaces, namely, liquid-vapor and liquid-liquid interfaces. The method requires no controlled fluid flow, is independent of conservative interactions between particle and interface, and resolves distance dependent friction to within a fraction of the particle radius. Near the liquid-vapor interface, the friction decreases below that in the bulk, corresponding to predictions of a "perfect-slip" surface.


Hypocretin neuron-specific transcriptome profiling identifies the sleep modulator Kcnh4a.
Sleep has been conserved throughout evolution; however, the molecular and neuronal mechanisms of sleep are largely unknown. The hypothalamic hypocretin/orexin (Hcrt) neurons regulate sleep\wake states, feeding, stress, and reward. To elucidate the mechanism that enables these various functions and to identify sleep regulators, we combined fluorescence cell sorting and RNA-seq in hcrt:EGFP zebrafish. Dozens of Hcrt-neuron-specific transcripts were identified and comprehensive high-resolution imaging revealed gene-specific localization in all or subsets of Hcrt neurons. Clusters of Hcrt-neuron-specific genes are predicted to be regulated by shared transcription factors. These findings show that Hcrt neurons are heterogeneous and that integrative molecular mechanisms orchestrate their diverse functions. The voltage-gated potassium channel Kcnh4a, which is expressed in all Hcrt neurons, was silenced by the CRISPR-mediated gene inactivation system. The mutant kcnh4a (kcnh4a(-/-)) larvae showed reduced sleep time and consolidation, specifically during the night, suggesting that Kcnh4a regulates sleep.


Electromyographic (EMG) normalization method for cycle fatigue protocols.
To determine the most effective electromyographic (EMG) normalization method for cycling fatigue protocols. Ten healthy subjects performed two 5-s isometric knee extension maximal voluntary contractions (MVC) at a knee joint angle of 60 degrees, two fixed cycle pedal contraction at knee joint angles of 60 degrees (60 degrees A) and 108 degrees (108 degrees A), and a dynamic single maximal revolution of a cycle pedal (1REV). Integrated EMG (IEMG) data were recorded for all contractions and power output recorded during MVC and 1REV. Mean IEMG for MVC was significantly (P < 0.01) greater than 60 degrees C, 108 degrees C, and 1REV. There were no significant differences between MVC and 1REV power output/EMG relationship. MVC will record a higher IEMG than 60 degrees A, 108 degrees A, and 1REV. As IEMG was greatest during MVC, and the relationship between IEMG and power output was not different between MVC and 1REV, normalization against maximal possible recruitment potential is most likely during MVC.


Hematology of chicks experiencing marginal vitamin B6 deficiency.
An economical vitamin B6-deficient ration, which was palatable to broiler chickens, was prepared and fed to 1-day-old broiler chicks. The experimental ration was a glucose-soybean meal diet. Vitamin B6 was removed by washing the soybean meal with water. Microbiological analysis revealed that the washed ration contained .45 mg vitamin B6 activity/kg. Experimental rations were formulated to contain .5, 1.0, and 3.0 mg added pyridoxine HCl/kg of ration. These supplemental levels produced total pyridoxine concentrations, as assayed, of .95, 1.48, and 3.18 mg pyridoxine HCl activity/kg of diet. Chicks were grown to 7 weeks of age and characteristic B6 deficiency traits, including increased mortality, decreased body weight gain, and increased incidence of abnormal leg conformation, were quantitated or observed. An extensive hematological evaluation at 4 weeks of age indicated that this marginal B6 deficiency resulted in increased erythrocyte numbers, decreased mean corpuscular hemoglobin levels, and increased erythrocyte fragility. No changes in hematocrits, total hemoglobin level, intracellular hemoglobin concentration, or reticulocyte number were found. These results indicate that anemia did not occur in broilers experiencing a moderate vitamin B6 deficiency. The hematological condition is described as microcytic, normochromic polycythemia.


Rab34 small GTPase is required for Hedgehog signaling and an early step of ciliary vesicle formation in mouse.
The primary cilium is a microtubule-based organelle that protrudes from the cell surface and plays essential roles in embryonic development. Ciliogenesis begins with the successive fusion of preciliary vesicles to form ciliary vesicles, which then dock onto the distal end of the mother centriole. Rab proteins have been linked to cilia formation in cultured cells, but not yet in vivo In the present study, we demonstrate that endocytic recycling protein Rab34 localizes to cilia, and that its mutation results in significant decrease of ciliogenesis in both cultured cells and mice. Rab34 is required for the successive fusion of preciliary vesicles to generate ciliary vesicles and for the migration of the mother centriole from perinuclear region to plasma membrane. We also show that Rab34 mutant mice exhibit polydactyly, and cleft-lip and -palate. These phenotypes are consistent with observations that nonciliated Rab34 mutant cells fail to respond to Hedgehog signaling and that processing of full-length Gli3 to its C-terminally truncated form is reduced in Rab34 mutant embryos. Therefore, Rab34 is required for an early step of ciliary vesicle formation and Hh signaling in vivo This article has an associated First Person interview with the first author of the paper.


Surgical Versus Medical Treatment for Diabetic Macular Edema: A Review.
We aimed to compare the results of pars plana vitrectomy (PPV) with internal limiting membrane (ILM) peeling, an alternative therapeutic strategy, with those of medical treatment for chronic macular edema. We conducted a review of the literature on the microscopic, anatomical, and functional reasons for performing PPV with ILM peeling in patients with diabetic macular edema (DME). We searched the PubMed database for articles published between 2000 and 2017. We used the medical subject heading "vitrectomy diabetic macular edema" and the keywords "diabetic macular edema", "internal limiting membrane peeling", "pars plana vitrectomy", "diabetic retinopathy", and "optical coherence tomography". Analysis of the literature revealed that cytokines, vascular endothelial growth factor, reactive oxygen species (ROS), and advanced glycation end-products (AGEs) play a unique role in DME. The vitreous cavity serves as a physiological reservoir for all inflammatory molecules. AGE receptors are localized at the footplates of Müller cells and the external limiting membrane (ELM). The footplates of Müller cells are in contact with the ILM, which suggests that they might be responsible for the structural damage (i.e., thickening) observed in the ILM of patients with DME. Therefore, PPV could allow a reduction of cytokines and pro-inflammatory molecules from the vitreous cavity. ILM peeling could eliminate not only the physical traction of a thickened structure, but also the natural reservoir of AGEs, ROS, and inflammatory molecules. PPV with ILM peeling is a surgical option that should be considered when treating patients with chronic DME.


CD8+ T cells in psoriatic lesions preferentially use T-cell receptor V beta 3 and/or V beta 13.1 genes.
Psoriasis is an inflammatory skin disorder characterized by epidermal keratinocyte hyperproliferation in association with a cellular infiltrate. There is evidence that activated T cells play a role in psoriatic plaque formation. We examined the T-cell receptor beta-chain variable gene segment (V beta) use of epidermal T cells in shave biopsies of psoriatic lesions. Our results show increased expression of V beta 3 and/or V beta 13.1 messages in the CD8+, but not CD4+, T cells in the lesions of a majority of patients studied. Sequence analysis of complementarity-determining region 3 (CDR3) of these two V beta genes from the skin demonstrated monoclonality or marked oligoclonality. A second biopsy from the same or different lesions, performed 3.5-8 months later in four patients, again revealed increased V beta 3 and/or V beta 13.1 expression and clonality. Moreover, in three of the four patients, the same V beta CDR3 rearrangement was found in both biopsies, although there was no V beta CDR3 homology between patients. In two patients in which V beta 3 and/or V beta 13.1 was not increased, an increase in V beta 17 gene use and clonality was found. The clonality of V beta sequence data indicates these cells are recruited and expanded in situ. The persistence of V beta 3-and/or V beta 13.1-bearing CD8+ T cells in lesions that did not undergo resolution suggests their role as effector cells rather than as regulatory cells.


Relative hypermetropia: a conception of Donders fallen into oblivion.
Donders created 2 triads in connection with hypermetropia. The first is well known: Ht = Hm + H1. The second is a division into facultative, relative, and absolute hypermetropia. In present ophthalmic literature the second triad is confused with the first; the important member called relative hypermetropia is not even mentioned. I tried to find the cause of this misunderstanding. In my opinion the second triad--in the form Hess had given to it--should be placed among the basic informations of ophthalmology.


Surgical management of syringomyelia: a five year experience in the era of magnetic resonance imaging.
We summarize our experience with 59 consecutive surgically managed cases of syringomyelia (SM) over a 5 year period. All cases had magnetic resonance imaging (MRI) preoperatively and postoperatively. Twenty-eight patients presented with SM and the adult Chiari (Chiari I) malformation (SM-ACM), 6 patients had post traumatic syrinxes, 14 patients had syrinxes associated with an intramedullary neoplasm, 3 patients had syringomyelia associated with spinal arachnoiditis and 8 patients had idiopathic syringomyelia. Holocord syrinxes were more often associated with SM-ACM, while focal syrinxes were associated with posttraumatic, spinal arachnoiditis and neoplastic conditions. In all cases with neoplasms the MRI revealed parenchymal intramedullary signal abnormalities in addition to the syrinx cavity. Posterior fossa decompression with obex plugging (the Gardner operation) was the procedure of choice for SM-ACM and for idiopathic holocord syringomyelia. Exploration and drainage of the syrinx with or without shunting was carried out mainly for focal syrinxes associated with trauma and neoplasm. Patients with SM-ACM responded well to posterior fossa decompression with satisfactory results in 24/28 patients. Idiopathic SM is probably a forme fruste of SM-ACM and when treated with the Gardner procedure showed good results in all 8 patients. Posttraumatic and neoplastic SM had the least predictable results with surgery. The disappearance of the syrinx on postoperative MRI correlated well with a good surgical outcome.


Refractory spontaneous hypoglycaemia: a diagnostic conundrum
A frail 79-year-old lady with dementia presented with a 2-year history of frequent falls. Recurrent hypoglycaemic episodes were diagnosed and treated with continuous glucose infusion in multiple hospital admissions. Hypoadrenalism and hypothyroidism were ruled out. Whilst hypoglycaemic (blood glucose 1.6 mmol/L), both plasma C-peptide and proinsulin concentrations, were inappropriately elevated at 4210 pmol/L (174–960) and >200 pmol/L (0–7) respectively with plasma insulin suppressed at 12 pmol/L (0–180). Whilst reported cases of proinsulinoma are typically pancreatic in origin, radiological investigations of the pancreas in this patient did not identify abnormalities. Unexpectedly contrast CT identified a heterogeneously enhancing mass (6.6 cm) at the lower pole of the left kidney consistent with renal cell carcinoma. Non-islet cell tumour-induced hypoglycaemia has been associated with renal malignancy; however, a serum IGF2:IGF1 ratio measured at <10 effectively excludes this diagnosis. Concomitantly on the CT, extensive peripherally enhancing heterogeneous mass lesions in the liver were identified, the largest measuring 12 cm. A palliative approach was taken due to multiple comorbidities. On post-mortem, the kidney lesion was confirmed as clear cell renal carcinoma, whilst the liver lesions were identified as proinsulin-secreting neuroendocrine tumours. In conclusion, the diagnosis of proinsulinoma can be missed if plasma proinsulin concentration is not measured at the time of hypoglycaemia. In this case, the plasma insulin:C-peptide ratio was too high to be accounted for by the faster relative clearance of insulin and was due to proinsulin cross-reactivity in the C-peptide assay. In addition, the concomitant malignancy proved to be a challenging red herring. Learning points: •• Even in non-diabetics, hypoglycaemia needs to be excluded in a setting of frequent falls. Insulin- or proinsulinsecreting tumours are potentially curable causes. •• Whilst investigating spontaneous hypoglycaemia, if plasma insulin concentration is appropriate for the hypoglycaemia, it is prudent to check proinsulin concentrations during the hypoglycaemic episode. •• Proinsulin cross-reacts variably with C-peptide and insulin assays; the effect is method dependent. In this case, the discrepancy between the insulin and C-peptide concentrations was too great to be accounted for by the faster relative clearance of insulin, raising the suspicion of assay interference. The C-peptide assay in question (Diasorin liaison) has been shown to be 100% cross reactive with proinsulin based on spiking studies with a proinsulin reference preparation. •• Whilst reported cases of proinsulinoma and 99% of insulinomas are of pancreatic origin, conventional imaging studies (CT, MRI or ultrasound) fail to detect neuroendocrine tumours <1 cm in 50% of cases. •• The concomitant renal mass identified radiologically proved to be a red herring. •• In view of the rarity of proinsulinoma, no conclusive association with renal cell carcinoma can be established.


A multifunction and programmable computerized barostat.
A new computerized barostat allowing digestive tone measurement but also programmed distensions is described. The apparatus consists of a pump, driven by a computer, which injects or aspirates air in a flacid bag placed in a part of the gastrointestinal tract when relaxation or contraction occurs. Several parameters can be programmed for tone measurement: mean pressure, window pressure and air flow rate. Isovolumic or isobaric distensions can be performed using three modes: linear, stepwise or step by step with intermediate deflation. The height (mL or mmHg) of the steps and the duration of the distensions and the deflations can also be programmed. Three automatic and one manual safety systems are included. In comparison with previously described barostats the main feature of this apparatus is that it is entirely computerized and can be used for programmed distensions.


Early life environmental predictors of asthma age-of-onset.
Prevention strategies that delay the onset of asthma may improve clinical outcomes. To identify early life environmental exposures associated with asthma age-of-onset and potential genetic modifiers of these exposures, we studied 1085 subjects with physician-diagnosed asthma and disease onset at or after age two. Subjects reported retrospectively on their exposure to 17 environmental factors before the age of two. The presence of individual or combinations of these early life exposures was then tested for association with variation in asthma age-of-onset. For exposures significantly associated with age-of-onset, we tested if 26 single nucleotide polymorphisms (SNP) with an established association with allergic disease significantly modified the effect of the exposure. Five environmental exposures were significantly associated with variation in asthma age-of-onset after correction for multiple testing: carpet at home (P = 6 × 10(-5)), a serious chest illness (P = 10(-4)), father a cigarette smoker (P = 6 × 10(-4)) and direct exposure to father's smoking (P = 3 × 10(-4)). Individuals with early childhood asthma onset, between the ages of two and six, were 1.4-fold (CI 1.1-1.9) more likely to report having lived in a house with carpet and 2.1-fold (CI 1.3-3.5) more likely to report suffering a serious chest illness before the age of two, than asthmatics with later disease onset. We further found these individual risks to increase to 3.2-fold (CI 1.7-6.0) if carpet exposure and suffering a serious chest illness co-occurred before age two. Paternal smoking exposures were less likely to be reported by asthmatics with early when compared to later disease onset (OR 0.5, CI 0.3-0.7). There were no significant SNP interactions with these environmental exposures after correction for multiple testing. Our results suggest that disease onset in individuals at a high-risk of developing asthma can potentially be delayed by avoiding exposure to carpet at home and preventing serious chest illnesses during the first 2 years of life.


Independent active contraction of extraocular muscle compartments.
Intramuscular innervation of horizontal rectus extraocular muscle (EOMs) is segregated into superior and inferior (transverse) compartments, whereas all EOMs are also divided into global (GL) and orbital (OL) layers with scleral and pulley insertions, respectively. Mechanical independence between both types of compartments has been demonstrated during passive tensile loading. We examined coupling between EOM compartments during active, ex vivo contraction. Fresh bovine EOMs were removed, and one compartment of each was coated with hydrophobic petrolatum. Contraction of the uncoated compartment was induced by immersion in a solution of 50 mM CaCl2 at 38°C labeled with sodium fluorescein dye, whereas tensions in both compartments were monitored by strain gauges. Control experiments omitted petrolatum so that the entire EOM contracted. After physiological experiments, EOMs were sectioned transversely to demonstrate specificity of CaCl2 permeation by yellow fluorescence dye excited by blue light. In control experiments without petrolatum, both transverse and GL and OL compartments contracted similarly. Selective compartmental omission of petrolatum caused markedly independent compartmental contraction whether measured at the GL or the OL insertions or for transverse compartments at the scleral insertion. Although some CaCl2 spread occurred, mean (±SD) tension in the coated compartments averaged only 10.5 ± 3.3% and 6.0 ± 1.5% in GL/OL and transverse compartments, respectively relative to uncoated compartments. Fluorescein penetration confirmed selective CaCl2 permeation. These data confirm passive tensile findings of mechanical independence of EOM compartments and extend results to active contraction. EOMs behave actively as if composed of mechanically independent parallel fiber bundles having different insertional targets, consistent with the active pulley and transverse compartmental hypotheses.


Premenopausal levels of circulating insulin-like growth factor I and the risk of postmenopausal breast cancer.
Increased levels of insulin-like growth factor I (IGF-I) may directly stimulate breast cell proliferation and promote growth and survival of transformed cells. Higher levels of IGF-I have been associated with increased risk of premenopausal breast cancer but not postmenopausal breast cancer. We investigated whether circulating levels of IGF-I prior to menopause are associated with breast cancer diagnosed after menopause in a population-based nested case-control study. Female cohort participants were enrolled in 1974 (n = 15,192) and 1989 (n = 18,724) and blood was drawn. Cases were women diagnosed with primary breast cancer at ages > or =50, of whom 152 were premenopausal at blood draw. One control was matched to each case on cohort participation, age, ethnic group, menopausal status and date of blood draw. Levels of IGF-I and IGF binding protein 3 (IGFBP-3) were measured using enzyme-linked immunoabsorbent assays. The association between IGF-I and breast cancer was determined using conditional logistic regression, adjusting for IGFBP-3. IGF-I levels decreased with age (p = 0.0001). Prior to age-stratification, IGF-I levels neither measured before nor after menopause were associated with postmenopausal breast cancer. After age-stratification, associations were suggested in the youngest premenopausal age group (upper vs. lowest third: odds ratio (OR) = 5.31, 95% confidence intervals (CI) = 0.85-33.13; p trend = 0.06) and oldest postmenopausal age group (upper vs. lowest third: OR = 3.41, 95% CI = 0.66-17.71; p trend = 0.13). The association between circulating levels of IGF-I and postmenopausal breast cancer risk may be modified by age. Increased levels of circulating IGF-I may be of particular interest in the younger premenopausal women and older postmenopausal women. Age-stratification should be undertaken in larger investigations of IGF-I levels as predictors of postmenopausal breast cancer.


Autologous Whole-Blood Versus Corticosteroid Local Injection in Treatment of Plantar Fasciitis: A Randomized Single Blind Placebo-Controlled Study.
Autologous whole-blood intralesional injection has attracted interest as a possible means of treatment for chronic plantar fasciitis. We contrasted its effectiveness with that of corticosteroids, which have a longer history of success in treating tendinopathies such as plantar fasciitis. In order to monitor the disease's progress naturally, we also compared them with a placebo. Sixty clinically diagnosed patients were taken up for intralesional injection of autologous whole blood (AWB), corticosteroid, and normal saline as placebo mixed with 2 mL of lignocaine after dividing them into three groups. Two doses were given and followed up in the third, sixth, and 12th weeks. The evaluation was done according to the visual analog scale (VAS) and the number of tablets of paracetamol (PCM) 500 mg consumed for the following period. When compared to the placebo group in the third, sixth, and 12th weeks, the corticosteroid group exhibited a significant improvement with a p<0.001 in the VAS score, whereas the autologous whole blood group showed no meaningful difference. When compared to the placebo group at the sixth and 12th weeks, the AWB group's VAS score showed a statistically significant difference with a p>0.001. At each follow-up, the placebo group consumed more analgesics than the corticosteroid group, with a p<0.001. Only in the third week of follow-up did AWB demonstrate a statistically significant difference in PCM consumption as compared to the corticosteroid group. Statistically significant improvement was seen in both the AWB group and corticosteroid group as compared to the placebo group. The corticosteroid group achieved earlier and superior relief of pain while AWB had a longer lasting effect. Therefore, study results indicate almost similar results in short-term studies.


Facile Synthesis of Novel Coumarin Derivatives, Antimicrobial Analysis, Enzyme Assay, Docking Study, ADMET Prediction and Toxicity Study.
The work reports the synthesis under solvent-free condition using the ionic liquid [Et₃NH][HSO₄] as a catalyst of fifteen novel 3-((dicyclohexylamino)(substituted phenyl/heteryl)-methyl)-4-hydroxy-2H-chromen-2-onederivatives 4a-o as potential antimicrobial agents. The structures of the synthesized compounds were confirmed by IR, ¹H-NMR, 13C-NMR, mass spectral studies and elemental analyses. All the synthesized compounds were evaluated for their in vitro antifungal and antibacterial activity. The compound 4k bearing 4-hydroxy-3-ethoxy group on the phenyl ring was found to be the most active antifungal agent. The compound 4e bearing a 2,4-difluoro group on the phenyl ring was found to be the most active antibacterial agent. The mode of action of the most promising antifungal compound 4k was established by an ergosterol extraction and quantitation assay. From the assay it was found that 4k acts by inhibition of ergosterol biosynthesis in C. albicans. Molecular docking studies revealed a highly spontaneous binding ability of the tested compounds to the active site of lanosterol 14α-demethylase, which suggests that the tested compounds inhibit the synthesis of this enzyme. The synthesized compounds were analyzed for in silico ADMET properties to establish oral drug like behavior and showed satisfactory results. To establish the antimicrobial selectivity and safety, the most active compounds 4e and 4k were further tested for cytotoxicity against human cancer cell line HeLa and were found to be non-cytotoxic in nature. An in vivo acute oral toxicity study was also performed for the most active compounds 4e and 4k and results indicated that the compounds are non-toxic.


[Splenic marginal zone lymphoma].
Splenic marginal zone lymphoma (SMZL) is a rare B-cell lymphoma, well defined according to the 2016 WHO classification of tumors of the hematopoietic tissue, which can raise diagnostic problems. We report the case of a 72-year old patient, hospitalised for exploration of a massive splenomegaly confirmed by computed tomography (CT). Laboratory tests were normal. The patient underwent splenectomy. Microscopic examination of the surgical specimen showed widespread lymphomatous proliferation in small cells associated with CD20 marker. CD 5 and CD 43 were negative. The diagnosis of SMZL was retained. SMZL accounts for less than 2% of non-Hodgkin lymphomas. It affects subjects older than 50 years, and it is usually characterized by the presence of a massive splenomegaly without lymphadenopathies. Blood count shows inconsistent presence of villous lymphocytes in three quarters of cases. The diagnosis is essentially based on anatomopathological evaluation and shows constant nodular involvement or sometimes diffuse involvement of white pulp of the splenic parenchyma.Tumor cells are small in size with B lymphoid markers expression: CD19, CD20, CD22, CD79. They are negative for CD5, CD10, cyclin D1 and CD43. No specific cytogenetic abnormality in SMZL has been identified. SMZL is an indolent lymphoma whose treatment is not codified, depending on prognostic factors. Mortality is related to the risk of its transformation to large cell lymphoma.


Chicken dystrophy. The geometry of the transverse tubules.
The pectoral muscle of chickens afflicted with muscular dystrophy, when examined with the electron microscope, contains a) numerous, often quite large vesicles with and without caveolar evaginations, b) tubules with caveolar evaginations, and c) tubular networks. We have demonstrated that all these structures are derivatives of the transverse tubles as revealed by tracer studies and freeze-fracture complementary replicas. The membranes of transverse tubular origin show a small number of intramembranous particles on both P and E faces with no complementary geometry. The membranes of the free sacrcoplasmic reticulum (SR) and the junctional SR of normal and dystrophic muscle appear identical in complementary freeze-fracture replicas. Vesicles that carry only a small number of particles on both E and P faces exposed by freeze-fracturing in isolated SR preparations can be taken as presumptive evidence and serve as a morphologic marker for transverse tubular origin of such vesicles when mitochondrial and lysosomal contamination has been excluded.


Changing ideas on antiviral agents for respiratory infection.
Since the early 1950s it has been shown that some nontoxic antiviral substances, given either orally or intransally, can inhibit the growth of respiratory viruses and their clinical effects. The next targets of research are the improvement of present methods of drug administration and the exploration of the possibility of giving drugs in combination. The most potent antiviral agents available should be used to establish whether drugs given when symptoms commence can have a significant effect, i.e., whether antiviral therapy is possible.


Acute renal failure following snakebite.
Eight patients with acute renal failure following snakebite were studied. Intravascular hemolysis and disseminated intravascular coagulation contributed to the development of acute renal failure in 6 patients. Direct nephrotoxicity causing acute renal failure is postulated in 2 patients, 1 of whom also revealed evidence of mild, disseminated intravascular coagulation. Three patients had histopathological lesions of acute symmetrical cortical necrosis and 3 had acute tubular necrosis. In 1 patient with acute tubular necrosis, in whom direct nephrotoxicity seemed to be responsible for renal failure, the striking histological feature was a uniform debasement and disappearance of tubular epithelium. In 2 patients with a clinical course of acute tubular necrosis, histological lesions could not be documented. All the 5 patients with acute tubular necrosis regained full recovery of renal function, 3 of them with the help of dialysis and 2 with conservative management. None of the 3 patients with acute cortical necrosis survived in spite of intermittent dialysis therapy.


Rectosigmoid (colonic) xanthoma: a report of four cases and review of the literature.
Xanthomas of the colon appear to be rare lesions. Four cases are reported to bring wider attention to this entity. Slides, reports and clinical notes from all cases coded as colonic or rectal xanthoma or xanthelasma during the period 1 January 1993 to 1 January 2000 were reviewed. The clinicopathological features are described and integrated into a review of nine previously reported cases. The ages of the patients ranged from 51 to 62 years (2M, 2F). Three lesions were located in the sigmoid colon and one in the rectum. They were all incidental findings at colonoscopy. Grossly, they varied from cream to yellow papules/polyps, ranged in size from 1 to 4mm and were all solitary. Two patients had an associated rectosigmoid hyperplastic polyp. There were no associated cutaneous xanthomatous lesions. One patient was diabetic and the serum lipid levels were normal in all patients. Histologically, all cases consisted of aggregates of mucosal foamy macrophages but in one case there were also macrophages involving the muscularis mucosae. The macrophages were negative for mucin and cytokeratin but were positive for CD68. Colonic xanthomas do not appear to be the cause of lower gastrointestinal symptoms and are an incidental colonoscopic finding as well as at times being seen in association with colonic polyps.


Distress in orthopedic trainees and attending surgeons: a Canadian national survey.
Physician health is of increasing concern in health care systems. The purpose of this study was to determine the prevalence of distress among orthopedic surgeons and trainees and to identify factors associated with distress. Voluntary, anonymous online surveys were sent to attending orthopedic surgeons and orthopedic trainees across Canada. The survey for attending surgeons used the Expanded Physician Well-Being Index, and the survey for trainees used the Resident/Fellow Well-Being Index. Demographic information was also collected. To look for predictors of physician distress, we evaluated the relationship between respondents' classification as "distressed" and "not distressed" against demographic factors. In total, 1138 attending orthopedic surgeons and 493 orthopedic trainees were invited to complete the survey. The survey response rate was 31.2% for attending orthopedic surgeons and 24.3% for orthopedic trainees. Overall, 55.4% of attending surgeons and 40.0% of trainees screened positive for distress. Among both attending surgeons and trainees, having dependents was not a risk factor for distress, nor was gender. Practice location was not a risk factor for distress among attending surgeons. Attending surgeons who were classified as distressed had spent significantly fewer years in practice (median 11 yr) than those who were classified as "not distressed" (median 16 yr) (p = 0.004). We found a higher rate of distress among orthopedic surgeons than has been previously reported. The distress rate among orthopedic trainees in this population is similar to that reported in other international publications, although self-reported rates of burnout were higher. The findings from this study may indicate a need for continuing research to determine intrinsic and extrinsic risk factors for distress among orthopedic surgeons and trainees and for the evaluation of prescriptive, evidence-based initiatives to address this crisis.


Subcutaneous perfusion of tumor necrosis factor induces local proliferation of fibroblasts, capillaries, and epidermal cells, or massive tissue necrosis.
Mouse recombinant tumor necrosis factor (TNF) (or its solvent alone as a control) was administered subcutaneously to mice by a cannula connected to an osmotic minipump. Perfusion at a rate of 35 ng/hr for seven days induced the formation of a tissue mass composed mainly of fibroblasts, collagen, and capillaries. Necrosis (apoptosis) of isolated fibroblasts was observed. Polymorphonuclear leukocytes were abundant after three to four days of perfusion but were absent later. The covering epidermis showed a hyperplastic reaction associated with necrosis of isolated keratinocytes. Perfusion at a rate of 170 ng/hr led, after four to five days, to a massive necrosis. Necrosis was completely prevented by rabbit anti-TNF IgG but not by anti-LPS IgG, irradiation, or administration of indomethacin.


Isolated limb perfusion: the European experience.
Isolated limb perfusion (ILP) is a method of cancer treatment allowing the administration of high doses of anticancer agents in a limb surgically isolated from systemic circulation. By using continuous leakage monitoring and using the drug melphalan, a high complete remission rate is obtained in patients with melanoma. In patients with sarcomas, ILP with tumor necrosis factor and melphalan represents a neoadjuvant treatment for limb-sparing surgery. This treatment is the first demonstration of an active anti-angiogenic regimen in the clinic.


Influence of CYP2C9 and VKORC1 polymorphisms on warfarin and acenocoumarol in a sample of Lebanese people.
The authors assessed the impact of CYP2C9*2, CYP2C9*3, and/or VKORC1-1639G>A/1173C>T single-nucleotide polymorphisms on oral anticoagulants in a Lebanese population. This study recruited 231 Lebanese participants on long-term warfarin or acenocoumarol maintenance therapy with an international normalized ratio (INR) monitored at the American University of Beirut Medical Center. CYP2C9 and VKORC1 variant alleles were screened by real-time PCR. Plasma R- and S-warfarin and R- and S-acenocoumarol levels were assayed using high-performance liquid chromatography. The variant allele frequencies of CYP2C9*2, CYP2C9*3, and VKORC1 -1639G>A/1173C>T were 15.4%, 7.8%, and 52.4%, respectively. Fifty-five participants were excluded from analysis because of nontherapeutic INR values at recruitment, leaving 43 participants taking warfarin and 133 taking acenocoumarol. There was a significant decrease in the weekly maintenance dose of both drugs with CYP2C9 and VKORC1 variants when compared with wild-type patients. CYP2C9*2 had the least impact on the response to both drugs. The concentrations of R- and S-warfarin in plasma were significantly correlated with CYP2C9 genotypes. For acenocoumarol, time to reach target INR was more prolonged in patients carrying any CYP2C9 variant allele but failed to reach statistical significance because of low numbers of patients. There was no association between allelic variants and bleeding events. This is the first pharmacogenetic study of oral anticoagulants in Arabs. The authors showed that both CYP2C9 and VKORC1 polymorphisms are common in Lebanon and influence warfarin and acenocoumarol dose requirements, with the CYP2C9*2 polymorphism having less effect on acenocoumarol, the most commonly used oral anticoagulant in Lebanon.


The incidence, treatment and prognosis of cervical carcinoma in young women: a retrospective analysis of 4,975 cases in Japan.
To determine the clinical characteristics of patients (young women) with cervical carcinoma aged less than 35 years. Data from patients who were treated for cervical carcinomas from 1990 to 2000 in the Kinki District were retrospectively investigated for clinical stage, histologic type, treatment procedure and prognosis. Of a total of 4,975 cases, 441 patients were aged less than 35 years old. The incidence of cervical carcinoma in these women was 7.9% from 1990 to 1995, 9.1% from 1996 to 2000, and 9.5% from 2001 to 2005. FIGO Stage I included 374 cases, followed by, 49 in Stage II, 11 in Stage III, and seven in Stage IV. Squamous cell carcinoma incidence was 80.7% and non-squamous cell carcinoma incidence was 19.3%. Several types of surgery were performed in patients with Stage I and II, while patients with Stage III and IV were treated with radiotherapy and/or chemotherapy without any type of surgery. In patients who underwent lymphadenectomy, 21.1% cases had nodal involvement. The 5-year survival rate was 95% for Stage I disease, 73% for Stage II, 68% for Stage III, and 19% for Stage IV. The incidence of cervical carcinoma in young women slightly increased from 1990 to 2005. The prognosis of cervical carcinoma tends to be better in young women than in older patients, especially in Stage III disease.


Comparison on hypoglycemic and antioxidant activities of the fresh and dried Portulaca oleracea L. in insulin-resistant HepG2 cells and streptozotocin-induced C57BL/6J diabetic mice.
Fresh Portulaca oleracea L. (family: Portulacaceae; POL) has been used as a folk medicine for the treatment of diabetes mellitus for a long time. More bioactive components with higher activity could be retained in fresh medicinal herbs compared to the dried ones. The present study was conducted to compare different antidiabetic activity between fresh and dried POL, including hypoglycemic and antioxidant activities both in vivo and in vitro. Furthermore, in order to explore which components were responsible for the antidiabetic activity, the difference on chemical components between fresh and dried POL was analyzed and compared. Insulin-resistant HepG2 cells induced by insulin were used to evaluate the promoting effect of the fresh and dried POL on glucose utilization in vitro. Streptozotocin (STZ)-induced C57BL/6J diabetic mice were used to compare the differences on hypoglycemic and antioxidant activities of fresh and dried POL, including the fasting blood glucose, glucose tolerance, serum insulin level, malondialdehyde (MDA) level and superoxide dismutase (SOD) activity in vivo. UPLC/Q-TOF-MS method was performed to analyze the difference of antidiabetic components between fresh and dried POL. Compared with the dried POL extract, the fresh POL extract significantly increased the consumption of extracellular glucose in insulin-resistant HepG2 cells (P<0.05). In STZ-induced C57BL/6J diabetic mice, both fresh and dried extracts decreased markedly the fasting blood glucose (FBG) levels, and improved significantly oral glucose tolerance test (OGTT), as well as enhanced significantly insulin secretion and antioxidative activities (P<0.05; P<0.01). Furthermore, the fresh extract showed stronger antidiabetic activity (P<0.05). The UPLC/Q-TOF-MS analysis results also revealed that the relative contents of polyphenols and alkaloids in the fresh herbs were more abundant than those in the dried POL. Our results indicated that both fresh and dried POL possessed antidiabetic activities, besides stronger activity was observed in the fresh herb. These findings provided evidence for the application and development of fresh POL in the treatment of diabetes mellitus.


The relationship between restrictive and repetitive behaviors in individuals with autism and obsessive compulsive symptoms in parents.
This study investigated the relationship between repetitive behaviors in individuals with autism and obsessive-compulsive behaviors in parents. We hypothesized that repetitive behaviors in probands with autism would be associated with increased obsessive-compulsive behaviors in parents in sporadic families (1 known case of autism per family and no known history of autism). Parents with clinically significant Y-BOCS scores were more likely to have a family history of obsessive-compulsive disorder. The empirically derived Autism Diagnostic Interview-R (ADI-R) factor, Insistence on Sameness, was positively correlated with obsessive-compulsive behaviors in parents. Further, when probands were grouped on the basis of parental Y-BOCS scores (clinically significant versus non-clinically significant), probands whose parents had clinically significant Y-BOCS scores had higher ADI-R Insistence on Sameness factor scores. The findings of the current study of sporadic families extend previous work that has shown an association between restrictive/repetitive behaviors in probands with autism and obsessive-compulsive features in parents.


miR-21a-5p Contributes to Porcine Hemagglutinating Encephalomyelitis Virus Proliferation via Targeting CASK-Interactive Protein1 In vivo and vitro.
Porcine hemagglutinating encephalomyelitis virus (PHEV) is a highly neurovirulent coronavirus that can cause nervous symptoms in piglets with muscle tremors, hind limb paralysis, and nystagmus. Whether some factors affect virus replication and proliferation had not been fully understood in the course of nerve damage caused by PHEV infection. In recent years, some reports suggested that miRNA might play a key regulatory role in viral infection. In this study, we found the miR-21a-5p is notably up-regulated in the brains of mice and N2a cells infected with PHEV, and it down-regulated the expression of CASK-interactive protein1 (Caskin1) by directly targeting the 3'-UTR of Caskin1 using a Dual-Luciferase reporter assay. The over-expression of miR-21a-5p or Caskin1 knockdown in the host significantly contributes to PHEV proliferation. Conversely, the silencing of miR-21a-5p by miR-21a-5p inhibitors suppressed the virus proliferation. Taken together, our results indicate that Caskin1 is the direct target gene of miR-21a-5p, and it is advantageous to virus proliferation by down-regulating Caskin1. These findings may help in the development of strategies for therapeutic applications.


[The removal efficiency of beta 2-microglobulin, alpha 1-microglobulin and alpha 1-acid glycoprotein by using dialyzers in the bloodstream and transference of endotoxin through membranes].
Retention of beta 2-microglobulin (beta 2-MG) has been indicated as one of the causes of hemodialysis-associated amyloidosis. Membranes with higher permeability (high-performance membrane) have recently been developed accordingly. Several dialyzers were tested by us in an attempt to study their removal efficiency of beta 2-MG and higher molecular substances of alpha 1-microglobulin (alpha 1-MG) and alpha 1-acid glycoprotein (alpha 1-AG), as well as transferring of endotoxin (ET) through membranes into bloodstream. The membranes subjected to our study were PMMA, CTA, PS, EVAL-C and CA. Removal efficiency of beta 2-MG through EVAL-C and CA was lower as compared with that through other membranes (P less than 0.05), while the EVAL-C showed relatively higher removal efficiency for alpha 1-MG and alpha 1-AG. Permeability for ET was not observed with all membranes studied herein.


Burst transcranial magnetic stimulation: which tinnitus characteristics influence the amount of transient tinnitus suppression?
Transcranial Magnetic Stimulation (TMS) is a method capable of temporarily suppressing tinnitus by delivering tonic or burst stimuli. Burst TMS has a high interindividual variability and low effect size. Tinnitus type and laterality, tinnitus-related distress, and tinnitus duration might contribute to this large individual variation. The effect of burst TMS on the auditory cortex in 100 male individuals is evaluated with coil placed over the auditory cortex. For unilateral tinnitus, this coil was placed contralaterally to the tinnitus, whilst for bilateral tinnitus the coil was placed over the right auditory cortex. The site of maximal tinnitus suppression is determined using 1-Hz stimulation with five pulses per burst (intensity of the stimulation set at 90% of the motor threshold). When tinnitus suppression is noted, the patients are asked to estimate the decrease in tinnitus in percentage using the numeric rating scale. The procedure is repeated with stimulations at 5, 10 and 20 Hz, each stimulation session consisting of 200 pulses. Results demonstrate that burst stimulation can decrease the perceived tinnitus intensity transiently in 57.83% of the patients. Patients with bilateral tinnitus respond better to burst TMS than patients with unilateral tinnitus and highly distressed patients presenting with unilateral pure tone tinnitus fail to bust TMS. Burst TMS modulates both unilateral and bilateral tinnitus, both high and low distress and both pure tones and narrow band tinnitus. However, the suppression effect is moderated by tinnitus type and laterality, tinnitus-related distress, and tinnitus duration.


From first symptoms to final diagnosis of Cushing's disease: experiences of 176 patients.
To obtain structured information on the diagnostic delay in patients with Cushing's disease (CD) from the patients perspective to provide leverage points for earlier diagnosis. The study includes 176 patients with ACTH-dependent CD who had received pituitary surgery completed a self-developed questionnaire on their symptomatology before the illness was diagnosed, the course and length of the diagnostic process, and the role of the involved health care professionals. Data were analyzed statistically. Answers in free text options were categorized and counted. The overall diagnostic process took 3.8±4.8 years (median 2 years), during which 4.6±3.8 (1-30) physicians were consulted, most frequently the family physician (FP; 83.0%). The presented symptoms were various and often vague, e.g. 'poor general condition' (at FPs), or very common in the field of the visited specialist (i.e. 'skin changes' at dermatologists). Women recognized the first CD symptoms themselves significantly more frequently than men, whereas physicians recognized CD symptoms significantly more frequently in males. A clear difficulty of diagnosing CD seems that patients describe isolated symptoms to the FP or the respective specialists according to their fields of specialization. As FPs are contacted most frequently, they should be trained to recognize the broad spectrum of CD symptoms, especially in female patients with weight gain, and initiate endocrinological referral.


Rapidly progressive atherosclerosis in aortocoronary saphenous vein grafts. Possible immune-mediated disease.
One hundred fifteen aortocoronary saphenous vein grafts from 100 consecutive reoperations were examined. Ninety-nine percent of the grafts contained diffuse concentric intimal fibroplasia. Forty-six percent of the grafts were atherosclerotic. The atherosclerosis was typically rapid in development, concentric, diffuse, without a fibrous cap, with numerous foam cells and inflammatory cells, including multinucleate giant cells, with erosion of the media by the inflammatory reaction. The histologic features of the grafts were similar to those reported in experimental models of immune-mediated atherosclerosis. On this basis we propose that aortocoronary saphenous vein graft atherosclerosis, and possibly intimal fibroplasia, may be an immune-mediated disease.


Optimization of walking in children.
Previous work demonstrated that adults naturally adopt a walking frequency to optimize physiological cost, symmetry, and stability. Furthermore, the optimal frequency is predictable using the force-driven harmonic oscillator (FDHO) model. However, no studies have established the developmental processes of optimization in children. Thus, the purposes of this study were to examine the predictability of the preferred stride frequency (PSF) and optimization features of 3- to 12-yr-old children using the FDHO model. Forty-five children and nine adults were measured for anthropometric data to calculate the predicted frequency. They later walked at three frequencies (PSF, PSF +25%, and PSF -25%) at a constant speed on a treadmill. The results indicated that the FDHO model was accurate in predicting the preferred frequency of children (prediction error < 0.07 s). We identified three stages of learning in the development of optimization: an early manifestation of sensitivity to resonant frequency, the subsequent development of ability to modulate walking frequency, and the final establishment of an adult optimization form at age seven. Our findings suggest that walking development may be determined by the dynamic cooperation of physiological, neural, and musculoskeletal systems with respect to the environmental context.


Initial testing (stage 1) of BAL101553, a novel tubulin binding agent, by the pediatric preclinical testing program.
BAL101553 is a highly water soluble prodrug of BAL27862 that arrests tumor cell proliferation and induces cell death in cancer cells through disruption of the microtubule network. In vitro BAL27862 demonstrated potent activity, with the median relative IC50 (rIC50 ) of 13.8 nM (range 5.4-25.2 nM). The in vitro activity of BAL27862 against the PPTP cell lines is distinctive from that previously described for vincristine. BAL101553 induced significant differences in EFS distribution compared to control in 16 of 30 (53%) solid tumor xenografts and in two of four (67%) of the evaluable ALL xenografts. No objective responses were observed.


Comparison between oscillometric and auscultatory methods of ambulatory blood pressure measurement in hemodialysis patients.
24-hour ambulatory blood pressure monitoring (ABPM) is commonly used in clinical and research practice. Different methods have been used in BP recording, cuff-oscillometric or Korotkoff sound, and validation studies during ABPM have been performed on general as well as hypertensive populations. Hemodialysis (HD) patients have a high percentage of complications, such as vascular diseases, and they are subject to hyperkinetic blood flows and abrupt body weight changes secondary to HD, which can invalidate BP recording. Therefore, we wanted to compare the 2 methods on an HD population. We performed 86 ABPMs on 44 patients (aged 60.8 +/- 17.2 years) by using a device capable of the simultaneous recording of oscillometric and auscultatory BP (A&D Takeda TM2421). The data obtained with the 2 different ABPM methods have been compared, and the differences between auscultatory and oscillometric determinations have been analyzed, as presented by Bland and Altman [1986]. The percentage of valid recordings was significantly higher with the oscillometric method than with the auscultatory method (93.6 +/- 11.3% vs. 71.7 +/- 17.04%, p < 0.001). 24-hour diastolic BP and night-time systolic BP were higher when recorded with the oscillometric method (DBP = 75.4 +/- 9.6 mmHg vs. 71.8 +/- 9.6 mmHg, p < 0.001, asleep SBP = 119.7+/-23.3 mmHg vs. 116.2 +/- 25.0 mmHg, p < 0.001), and the systolic night/day BP ratio was also higher(0.92 +/- 0.10vs.0.90 +/- 0.10, p < 0.001). Finally, the BP coefficient of variation ((SD/mean BP) x 100) was higher when auscultatory determinations were used (16.1 +/- 4.6 vs. 14.6 +/- 4.9). The limits of agreement between auscultatory and oscillometric BP determinations were for SBP = -6.44; 7.84 and for DBP = -3.66; 10.86. Differences between 24-hour oscillometric and auscultatory ABPM were reported in HD patients: the diastolic 24-hour and asleep systolic BP values and the systolic night/day ratio obtained with the oscillometric method were significantly higher. The higher coefficient of variation reported with the auscultatory method and the wider limits of agreement suggest that the 2 methods do not fully coincide and, in our opinion, the oscillometric method is preferable, due to the higher number of 24-hour valid measurements.


Auditory evoked potentials during propofol anaesthesia in man.
The effects of propofol on auditory evoked potentials were studied in nine patients undergoing otorhinolaryngology surgery. After recording of basal evoked potentials patients received propofol 2 mg kg-1 over 2-3 min for induction of anaesthesia. Potentials were recorded every 10 min (T1, T2, T3). During T1, T2, T3, the infusion rates of propofol for maintenance of anaesthesia were respectively 7, 5 and 3 mg kg-1 h-1 consecutively. Middle latency component was affected markedly. Brainstem waves latencies I, III, V were increased significantly, while amplitude waves I, III, V remained constant.


Magnetic Isolation, RNA Extraction, and Nanostring Analysis of Human Peripheral Blood Leukocytes Subsets.
Advancements in multiplexed molecular biology techniques have allowed for blood samples and specific circulating blood leukocytes to be a useful sample source when examining systemic changes associated with changes in body weight, muscle injury, disease onset/progression, and other common conditions. One gap in the current scientific knowledge relates to the impact of changes in individual leukocyte subsets on the overall systemic response. While many studies have published data related to changes observed in a mixed population of circulating leukocytes (i.e., whole blood sample), few studies have identified which cell(s) are responsible for the overall change. Since it is established that leukocyte subsets respond differently to various experimental challenges, it may be possible to gain new insight regarding overall biological state. This has application to a variety of health, nutrition, and exercise intervention models. Despite the need to examine changes in mRNA expression levels in individual leukocyte subsets they are not always easy to isolate and perform mRNA analysis on. In this report we describe a method to magnetically isolate, stabilize RNA, and analyze 800+ mRNA in a single sample analysis. Further we compared total leukocyte and leukocyte subset (i.e., granulocytes, monocytes, and T-cells) mRNA expression to better understand how subset changes contribute to overall response. Examination of subset responses may provide targets for future intervention studies. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Automated magnetic isolation of granulocytes, monocytes, and T-cells Basic Protocol 2: RNA extraction from magnetically sorted granulocytes, monocytes, and T-cells Basic Protocol 3: Nanostring analysis of RNA extracted from magnetically sorted granulocytes, monocytes, and T-cells.


Single-dose pharmacokinetics of methylphenidate extended-release multiple layer beads administered as intact capsule or sprinkles versus methylphenidate immediate-release tablets (Ritalin(®)) in healthy adult volunteers.
The purpose of this study was to evaluate the relative bioavailability and safety of a multilayer extended-release bead methylphenidate (MPH) hydrochloride 80 mg (MPH-MLR) capsule or sprinkles (37% immediate-release [IR]) versus MPH hydrochloride IR(Ritalin(®)) tablets, and to develop a pharmacokinetic (PK) model simulating MPH concentration-time data for different MPH-MLR dosage strengths. This was a single-center, randomized, open-label, three-period crossover study conducted in 26 fasted healthy adults (mean weight±standard deviation, 70.4±11.7 kg) assigned to single-dose oral MPH-MLR 80 mg capsule or sprinkles with applesauce, or Ritalin IR 25 mg (1×5 mg and 1×20 mg tablet) administered at 0, 4, and 8 hours. MPH-MLR 80 mg capsule and sprinkles were bioequivalent; ratios for maximum concentration (Cmax), area under plasma drug concentration versus time curve (AUC)0-t, and AUC0-inf were 1.04 (95% confidence interval [CI], 96.3-112.4), 0.99 (95% CI, 95.3-102.8), and 0.99 (95% CI, 95.4-103.0), respectively. MPH-MLR capsule/sprinkles produced highly comparable, biphasic profiles of plasma MPH concentrations characterized by rapid initial peak, followed by moderate decline until 5 hours postdose, and gradual increase until 7 hours postdose, culminating in an attenuated second peak. Based on 90% CIs, total systemic exposure to MPH-MLR 80 mg capsule/sprinkles was similar to that for Ritalin IR 25 mg three times daily, but marked differences in Cmax values indicated that MPH-MLR regimens were not bioequivalent to Ritalin. MPH Cmax and total systemic exposure over the first 4 hours postdose with MPH-MLR capsule/sprinkles was markedly higher than that associated with the first dose of Ritalin. All study drugs were safe and well tolerated. The PK modeling in adults suggested that differences in MPH pharmacokinetics between MPH-MLR and Ritalin are the result of dosage form design attributes and the associated absorption profiles of MPH. MPH-MLR 80 mg provides a long-acting biphasic pattern of plasma MPH concentrations with one less peak and trough than Ritalin IR.


Epididymal functions and their hormonal regulation.
The epididymis is a complex organ which maintains a specific intraluminal environment thought to be important for effecting sperm maturation in proximal regions and sperm storage in distal regions of the duct. The composition of the internal milieu is achieved both by transport between blood and lumen (and vice versa) and by synthesis and secretion into the lumen. Several low-molecular weight organic molecules achieve high concentration in the epididymal lumen, but their functions in the events of sperm maturation and storage still remain unclear. Metabolic processes occurring within epididymal tissue and the absorptive and secretory activity of the epididymal epithelium are regulated by androgens. The synthesis of some, but not all, secretory proteins is also androgen-dependent. In addition to androgens, other hormones and local testicular factors may influence epididymal function. There is now increasing evidence that epididymal-specific and androgen-dependent secretory proteins play a fundamental role in modifying the surface characteristics of sperm in preparation for the events of fertilization.


SP and KLF Transcription Factors in Digestive Physiology and Diseases.
Specificity proteins (SPs) and Krüppel-like factors (KLFs) belong to the family of transcription factors that contain conserved zinc finger domains involved in binding to target DNA sequences. Many of these proteins are expressed in different tissues and have distinct tissue-specific activities and functions. Studies have shown that SPs and KLFs regulate not only physiological processes such as growth, development, differentiation, proliferation, and embryogenesis, but pathogenesis of many diseases, including cancer and inflammatory disorders. Consistently, these proteins have been shown to regulate normal functions and pathobiology in the digestive system. We review recent findings on the tissue- and organ-specific functions of SPs and KLFs in the digestive system including the oral cavity, esophagus, stomach, small and large intestines, pancreas, and liver. We provide a list of agents under development to target these proteins.


Parental (non-)pain attending verbalizations moderate the relationship between child attention and memory bias for pain.
Children's negatively biased pain-related memories (i.e. recalling pain as being more intense or fearful than initially reported) have been recognized as a key factor in explaining child pain development. While mechanisms underlying children's pain memory development remain poorly understood, attention biases and parent language have been implicated in conceptual models. This study examined the association between child pain-related attention and memory biases and the moderating role of parental pain and non-pain attending verbalizations. Participants were 51 school children and one of their parents. Probability of initial fixation and gaze duration to pain were assessed using eye tracking methodology. Children performed a cold pressor task (CPT) and reported on experienced pain intensity and pain-related fear. A 3-minute parent-child interaction upon CPT completion allowed measurement of parental pain and non-pain attending verbalizations. Children's pain-related memories were elicited 2 weeks later. Findings indicated that the relationship between maintained attention to pain and fear memory bias was moderated by parental non-pain attending verbalizations such that higher gaze duration bias was positively associated with fear memory bias but only among children whose parents demonstrated low levels of non-pain attending verbalizations. The opposite pattern was observed for children whose parents showed high levels of non-pain attending verbalizations. No such effects were observed for child initial attention bias to pain, memory bias for pain and parental pain attending verbalizations. Findings highlight the importance of parental and child pain-related variables as well as their interaction in understanding negatively biased pain-related memories. This study on child pain memories is the first to highlight that characteristics of the social context, such as parental (non-)pain-related verbalizations, as well as factors related to the intra-individual experience of pain, such as child attention bias to pain, should be studied jointly, as they interact with each other in their effect on the emergence of negatively biased memories of painful events.


Spleen cell transfer induces T cell-dependent granulomas in tuberculous nude mice.
After IV infection with an attenuated strain of Mycobacterium bovis BCG, athymic nude mice produced a few hepatic granulomas; most of the other foci of bacterial parasitization lacked obvious cellular reactions. Nude mice BCG-infected four weeks earlier received spleen cells from normal euthymic littermates. After transient splenomegaly which peaked at about two weeks, numerous hepatic granulomas appeared by four weeks and the number of viable BCG decreased. The development of hepatic granulomas required viable spleen cells and depended on the spleen cell dose and corresponded with positive footpad reactions. The effective population seemed to be T cells, since they were sensitive to treatment with anti-brain associated theta antigen serum (antiBAT) plus complement, and were found in nylon-wool passed fractions. After transfer with sensitized spleen cells, adoptive immunity was observed, provided that the recipient BCG-infected nude mice had been splenectomized before transfer. AntiBAT plus complement treatment of the sensitized spleen cells decreased the granuloma-producing as well as antimycobacterial capacity. When BCG-infected nude mice were irradiated with 500 r of gamma ray, granuloma production was decreased after immune spleen cell transfer. Co-transfer of immune spleen cells with normal nude mouse bone marrow cells to BCG-infected irradiated nude recipients restored the granuloma-forming ability.


Superconductivity in Perovskite Ba1-xLnx(Bi0.20Pb0.80)O3-δ (Ln = La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu).
Solid solutions Ba1-xLnx(Bi0.20Pb0.80)O3-δ (Ln = La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu; 0.00 ≤ x ≲ 0.15) have been prepared under 850 °C. They all crystallize in space group P1 at room temperature. XPS data indicate that the valences are 5+ and 3+ for bismuth, 4+ and 2+ for lead, and 3+ or 4+ for lanthanide. Some of them are superconductors. The superconductive transition temperature Tczero decreases or remains constant with an increase of Ln in the sample when Ln = La, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu. However, Tczero first decreases, then increases, and finally decreases when Ln = Ce, Pr, which is due to the corresponding sample changes from hole-doped to electron-doped superconductors with an increase of Ce or Pr in the sample.


Digital subtraction angiography of the left ventricle.
Twenty-eight patients have been studied by left ventricular angiography and digital subtraction angiography, using either a peripheral venous injection (16) or a central venous injection (12) of contrast medium. Digital subtraction angiography compared favourably to angiography in the qualitative assessment of left ventricular size and function. Measurement of ejection fraction demonstrated a good correlation (r = 0.83) between digital subtraction angiography and left ventricular angiography. In the assessment of left ventricular segmental wall motion the use of a central venous injection, particularly in those patients with severely impaired ventricular function, proved superior to the use of a peripheral injection.


Comparison of casein micelles in raw and reconstituted skim milk.
During the manufacture of skim milk powder, many important alterations to the casein micelles occur. This study investigates the nature and cause of these alterations and their reversibility upon reconstitution of the powders in water. Samples of skim milk and powder were taken at different stages of commercial production of low-, medium-, and high-heat powders. The nature and composition of the casein micelles were analyzed using a variety of analytical techniques including photon correlation spectroscopy, transmission electron microscopy, turbidity, and protein electrophoresis. It was found that during heat treatment, whey proteins are denatured and become attached to the casein micelles, resulting in larger micelles and more turbid milk. The extent of whey protein attachment to the micelles is directly related to the severity of the heat treatment. It also appeared that whey proteins denatured during heat treatment may continue to attach to casein micelles during water removal (evaporation and spray-drying). The process of water removal causes casein and Ca in the serum to become increasingly associated with the micelles. This results in much larger, denser micelles, increasing the turbidity while decreasing the viscosity of the milk. During reconstitution, the native equilibrium between colloidal Ca and serum Ca is slowly reestablished. The reequilibration of the caseins and detachment of the whey proteins occur even more slowly. The rate of reequilibration does not appear to be influenced by shear or temperature in the range of 4 to 40 degrees C.


Scalings of domain wall energies in two dimensional Ising spin glasses.
We study domain wall energies of two dimensional spin glasses. The scaling of these energies depends on the model's distribution of quenched random couplings, falling into three different classes. The first class is associated with the exponent theta approximately -0.28; the other two classes have theta=0, as can be justified theoretically. In contrast to previous claims, we find that theta=0 does not indicate d=d(c)(l) but rather d< or =d(c)(l), where d(c)(l) is the lower critical dimension.


Phylogenetic mixtures on a single tree can mimic a tree of another topology.
Phylogenetic mixtures model the inhomogeneous molecular evolution commonly observed in data. The performance of phylogenetic reconstruction methods where the underlying data are generated by a mixture model has stimulated considerable recent debate. Much of the controversy stems from simulations of mixture model data on a given tree topology for which reconstruction algorithms output a tree of a different topology; these findings were held up to show the shortcomings of particular tree reconstruction methods. In so doing, the underlying assumption was that mixture model data on one topology can be distinguished from data evolved on an unmixed tree of another topology given enough data and the "correct" method. Here we show that this assumption can be false. For biologists, our results imply that, for example, the combined data from two genes whose phylogenetic trees differ only in terms of branch lengths can perfectly fit a tree of a different topology.


Synergistic tumoricidal effect between celecoxib and adenoviral-mediated delivery of mda-7 in human breast cancer cells.
Celecoxib, a selective cyclooxygenase 2 (COX-2) inhibitor, blocks growth and promotes apoptosis in breast cancer cells. The PI3K/Akt pathway is important in cell survival, and COX-2 and Akt might promote growth via a positive feedback loop. We have shown that adenoviral delivery of mda-7 (Ad-mda7) in breast cancer down-regulates Akt. We hypothesized that combining Ad-mda7 and celecoxib could mediate tumor suppression in COX-2 overexpressing breast cancer cells. Two COX-2 overexpressing human breast cancer cell lines (Her-18 and MDA-MB-436) were treated with celecoxib (20 micromol/L and 50 micromol/L) and Ad-mda7 (multiplicity of infection, 1000 and 2000 viral particles/cell). Adenovirus encoding the luciferase gene was used as a control. We assessed proliferation, cell cycle, apoptosis, prostaglandin E2 production, and changes in protein expression. Statistical analysis was performed by using the Student t test. Regardless of HER-2/neu status, cell growth was markedly inhibited by celecoxib, Ad-mda7, and the combination compared with controls. Celecoxib + Ad-mda7 showed a greater than additive increase in cell death compared with either monotherapy (P < .05) and resulted in cell cycle block and apoptosis (P < .05). Both cell lines showed decreased prostaglandin E2 production after combination treatment compared with controls (P < .05), with decreased expression of COX-2, Akt, and phosphorylated Akt (P < .05). Enhanced antitumor activity is achieved in breast cancer by combining celecoxib and Ad-mda7 regardless of HER-2/neu status. This occurs through inhibition of COX-2 expression and down-regulation of Akt. Combining Ad-mda7 with COX-2 inhibition provides a novel method of treatment in breast cancer.


Hedycaryol synthase in complex with nerolidol reveals terpene cyclase mechanism.
The biosynthesis of terpenes is catalysed by class I and II terpene cyclases. Here we present structural data from a class I hedycaryol synthase in complex with nerolidol, serving as a surrogate for the reaction intermediate nerolidyl diphosphate. This prefolded ligand allows mapping of the active site and hence the identification of a key carbonyl oxygen of Val179, a highly conserved helix break (G1/2) and its corresponding helix dipole. Stabilising the carbocation at the substrate's C1 position, these elements act in concert to catalyse the 1,10 ring closure, thereby exclusively generating the anti-Markovnikov product. The delineation of a general mechanistic scaffold was confirmed by site-specific mutations. This work serves as a basis for understanding carbocation chemistry in enzymatic reactions and should contribute to future application of these enzymes in organic synthesis.


HIF-1: an age-dependent regulator of lens cell proliferation.
The lens grows throughout life, and lens size is a major risk factor for nuclear and cortical cataracts. A previous study showed that the hypoxic environment around the lens suppressed lens growth in older rats. The present study was conducted to investigate the mechanism responsible for the age-dependent decline in lens cell proliferation. Transgenic mice expressing Cre recombinase in the lens were bred to mice containing floxed Hif1a alleles. Transgenic mice expressing oxygen insensitive forms of HIF-1alpha in lens epithelial cells were exposed to room air or 60% oxygen. Proliferation was measured by BrdU labeling and cell death by using the TUNEL assay. Morphology was assessed in histologic sections. HIF-1alpha and p27(KIP1) levels were determined by Western blot. The expression of HIF-regulated genes was assessed on microarrays. Lenses lacking Hif1a degenerated, precluding study in older animals. Breathing 60% oxygen reduced HIF-1alpha levels and HIF-1-regulated transcripts in lens epithelial cells from young and older lenses. Overexpression of oxygen-insensitive HIF-1alpha had no effect on lens size, but suppressed increased proliferation in response to oxygen. Systemic injection of the iron chelator, 1,10-phenanthroline prevented the degradation of HIF-1alpha and reduced oxygen-induced proliferation. Increasing oxygen decreased levels of p27(KIP1) in the epithelial cells of older mice, which was prevented by expressing oxygen-insensitive forms of HIF-1alpha. HIF-1alpha is present and HIF-1 is transcriptionally active throughout life, but suppresses growth only in older lenses. Maintaining elevated levels of p27(KIP1) in older lenses requires HIF-1. p27(KIP1) and other growth regulators may selectively suppress the proliferation of older lens epithelial cells.


Transcutaneous auricular vagus nerve stimulation attenuates inflammatory bowel disease in children: a proof-of-concept clinical trial.
Vagus nerve stimulation is an investigational anti-inflammatory therapy targeting the nervous system to modulate immune activity. This study evaluated the efficacy and safety of transcutaneous auricular VNS (ta-VNS) in patients with pediatric-onset Crohn's disease (CD) or ulcerative colitis (UC). Participants were 10-21 years of age with mild/moderate CD or UC and fecal calprotectin (FC) > 200 ug/g within 4 weeks of study entry. Subjects were randomized to receive either ta-VNS targeting the cymba conchae of the external left ear, or sham stimulation, of 5 min duration once daily for a 2-week period, followed by a cross over to the alternative stimulation for an additional 2 weeks. At week 4, all subjects received ta-VNS of 5 min duration twice daily until week 16. Primary study endpoints were clinical remission, and a ≥ 50% reduction in FC level from baseline to week 16. Heart rate variability measurements and patient-reported outcome questionnaires were completed during interval and week 16 assessments. Twenty-two subjects were enrolled and analyzed (10 CD, 12 UC). Six of 10 with CD had a wPCDAI > 12.5 and 6/12 with UC had a PUCAI > 10 at baseline, correlating to mild to moderate symptom activity. Among the 12 subjects with active symptomatic disease indices at baseline, clinical remission was achieved in 3/6 (50%) with CD and 2/6 (33%) with UC at week 16. Despite all subjects having FC levels ≥ 200 within 4 weeks of enrollment, five subjects (4 UC, 1 CD) had FC levels < 200 at the baseline visit and were excluded from the FC analysis. Of the remaining 17, median baseline FC was 907 µg/g (IQR 411-2,120). At week 16, 11/17 (64.7%) of those with baseline FC ≥ 200 had a ≥ 50% reduction in FC (95% CI 38.3-85.8). In the UC subjects, there was an 81% median reduction in FC vs baseline (833 µg/g; p = 0.03) while in the CD subjects, median reduction in FC at 16 weeks was 51% (357 µg/g; p = 0.09). There were no safety concerns. Noninvasive ta-VNS attenuated signs and symptoms in a pediatric cohort with mild to moderate inflammatory bowel disease. NCT03863704-Date of registration 3/4/2019.


[Analysis of changes in serological factors in the dose-increasing phase of allergic rhinitis rush immunotherapy].
Objective:To observe the changes of serological factors in the dose-increasing phase of allergic rhinitis and to explore the mechanism of short-term rush immunotherapy.Method:The clinical data of 110 patients with allergic rhinitis diagnosed by rush immunotherapy in our hospital were analyzed. According to the results of allergen test(sIgE), they were divided into simple dust mite allergy group and multiple allergen allergy groups. The serum of the patients were taken before the start of rush immunotherapy and the first day after the end of the dose-increasing phase.Serum total IgE and IgG before and after rush immunotherapy were detected by Beekman specific protein detector. The serum levels of IFN-γ,IL-12,IL-10,IgG1 and IgG4 were measured by ELISA before and after rush immunotherapy. The changes of them were compared and analyzed.Result:①Serum total IgE and IgG were increased after rush immunotherapy, and the change of serum IgG concentration in the simple dust mite allergy group was statistically significant(P<0.05).②The levels of serum IFN-γ, IL-12 and IL-10 increased after rush immunotherapy. The changes of IL-12 and IFN-γ in multiple allergen allergy groups were not significant before and after treatment, while the changes in other groups were statistically significant(P<0.05).③Serum total IgG1 and IgG4 were increased after rush immunotherapy. The changes of serum IgG4 in the simple dust mite allergy group were statistically significant(P<0.05). There was no statistically significant difference in serum IgG1.Conclusion:The dose-increasing phase of rush immunotherapy can stimulate the production of IgG antibodies and the changes of related serological factors in a short period of time, especially in patients with simple dust mite allergy group. It indicates that dose-increasing phase of rush immunotherapy may play a role by regulating immunoglobulin and adjusting the dynamic balance of Th1/Th2 factors.


Clopidogrel hypersensitivity syndrome with rash, fever, and neutropenia.
We report a case of a severe hypersensitivity syndrome reaction to clopidogrel that resolved with dechallenge and recurred on rechallenge. The reaction included neutropenia, rash, fever, tachycardia, nausea, and vomiting. This presentation is very similar to the frequently reported hypersensitivity syndrome reactions to the structurally similar drug ticlopidine and is very similar to a case involving clopidogrel reported previously in this Journal. We could find no other reports of similar cases related to clopidogrel.


Enantioseparation of chiral alcohols by complex formation and subsequent supercritical fluid extraction.
The very first application of supercritical fluid extraction (SFE) on enantioseparation of alcohols is discussed. Resolution of three chiral alcohols (trans-2-chloro-cyclohexanol, trans-2-bromo-cyclohexanol, and trans-2-iodo-cyclohexanol) were performed by partial complexation with (-)-O,O'-dibenzoyl-(2R,3R)-tartaric acid monohydrate (DBTA). DBTA formed diastereomeric complexes with all S,S-enantiomers stable enough to extract the unreacted alcohols with supercritical carbon dioxide. Resolution efficiency increased with the size of halogen substituents, and by the proper selection of molar ratio, pure (-)-R,R-trans-2-iodo-cyclohexanol (ee > 99%, yield: 39%) or (+)-S,S-trans-2-iodo-cyclohexanol (ee = 98%, yield: 8%) were prepared in one process step. Achieved resolution efficiency values were much higher in all resolution procedures than in any other known enantioseparation of these racemic compounds. The developed method offers an environmentally friendly, efficient alternative of currently applied resolution processes, also on a preparative scale.


MRCK ensures cortex-chromatin "social distancing" to enable egg spindle rotation.
During the second meiotic cell division, egg cells discard one set of chromatids to the polar body to produce a large haploid gamete. Meiotic spindle rotation is a critical step to ensure proper polar body extrusion. In this issue, Bourdais et al. (2023. J. Cell Biol.https://doi.org/10.1083/jcb.202211029) have identified MRCKβ as an essential kinase for efficient spindle rotation. MRCK activates cortical myosin II rings overlying the spindle to prevent the notoriously sticky interaction between the cell cortex and chromatin to facilitate spindle rotation. Furthermore, Bourdais et al. found that the same MRCK-myosin II pathway also operates in zygotes to promote parental genome unification.


Reactivity of a Tetra(o-tolyl)diborane(4) Dianion as a Diarylboryl Anion Equivalent.
Lithium and magnesium salts of tetra(o-tolyl)diborane(4) dianion, having B=B double bond character, were synthesized. It was clarified that the lithium salt of the dianion has a high-lying HOMO and a narrow HOMO-LUMO gap, which were perturbed by dissociation of Li+ cation, as judged by UV/Vis spectroscopy and DFT calculations. The lithium salt of the dianion reacted as two equivalents of a diarylboryl anion with CH2 Cl2 or S8 to give boryl-substituted products.