Domain-dependent modulation of insulin-induced AS160 phosphorylation and glucose uptake by Ca2+/calmodulin-dependent protein kinase II in L6 myotubes.
In skeletal muscle, the molecular mechanisms by which insulin stimulates glucose transport remains incompletely understood. Our study investigated the cellular dynamics of intracellular Ca(2+) mobilisation and Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) activation on insulin-induced skeletal muscle glucose transport. L6 myotubes were treated without or with insulin [100 nM] for 15 min and subsequently monitored for glucose uptake using isotope-labelled 2-deoxyglucose (I-2DOG), intracellular Ca(2+) (Ca(i)(2+)) release using Fluo-4AM and protein phosphorylation using Western blotting. Acute exposure of myotubes to insulin increased both Akt substrate-160 kDa (AS160) phosphorylation and I-2DOG uptake. Insulin concurrently increased Ca(i)(2+) and activated CaMKII. Exposing myotubes to either BAPTA/AM to sequester Ca(i)(2+) or KN-93 to inhibit CaMKII activity, decreased insulin-induced glucose uptake without affecting AS160 phosphorylation. On the other hand, blocking either calmodulin or the autoregulatory domain of CaMKII blocked the effect of insulin on both AS160 phosphorylation and glucose transport. Likewise, genetic knockdown of CaMKII in myotubes using siRNA completely abolished insulin-mediated glucose uptake. These results illustrate impairments in Ca(i)(2+) mobilisation and CaMKII activation are sufficient to negatively influence insulin-dependent glucose transport by L6 myotubes. Additionally, our results show for the first time that Ca(i)(2+) and domain-dependent CaMKII signalling differentially affect insulin-induced AS160 phosphorylation, and establish that Ca(2+) and CaMKII are components of the insulin signalling pathway in L6 myotubes.


Rare detection of phenotypically immature lymphocytes in Hashimoto thyroiditis and rheumatoid arthritis.
It has been suggested that recombination activating gene (RAG)-dependent revision of the immunoglobulin genes in germinal centres may contribute to local production of autoantibodies in Hashimoto thyroiditis (HT) and rheumatoid arthritis (RA). To test this hypothesis we examined HT and RA tissues for expression of RAG and terminal deoxynucleotidyl transferase (TdT) in situ. Paraffin-embedded tissues from 19 HT patients and from 20 RA patients were subjected to immunohistochemistry using TdT-specific antibodies. Expression of the RAGs was studied by in situ hybridisation. Tonsil sections were used as a control. Expression of TdT and RAGs was detected in extrafollicular lymphocytes in control tonsil sections. By contrast, only rare TdT-expressing cells were identified in 11 of 19 HT and in 2 of 20 RA samples. Germinal centre B-cells were consistently TdT- and RAG-negative. These results suggest that local RAG-dependent receptor revision in germinal centres is unlikely to contribute to production of autoantibodies in HT and RA. The presence of TdT-positive extrafollicular cells may represent an influx of immature cells in the context of chronic immune stimulation.


Short- or long-stem prosthesis for intramedullary bypass of proximal humeral fractures with severe metaphyseal bone loss: evaluation of primary stability in a biomechanical model.
Proximal humeral fractures with substantial metaphyseal comminution are challenging to treat. In the elderly with osteoporotic bone, arthroplasty sometimes remains the only valuable option; however, the minimally required length of stem fixation is not known. The aim of this study was to investigate the primary stability of cemented short- and long-stem prostheses with different intramedullary fracture bypass lengths. Osteoporotic composite bone models of the humerus (Synbone, Malans, Switzerland) with 3 different fracture levels (group A, 6 cm distal to surgical neck; group B, 7 cm distal to surgical neck; and group C, 8 cm distal to surgical neck) were prepared with a cemented standard short (S)- or long (L)-stem prosthesis and were tested for torque to failure. As a reference, we used models with intact bone (group R-O) and a short-stem prosthesis implanted at the surgical neck (group R-P). The radiographic bypass index (BI) was calculated before testing (fracture level to stem tip [in millimeters]/outer cortical diameter at fracture level [in millimeters]). The resulting BIs of each group were as follows: 1.7 in group A-S, 3.4 in group A-L, 1.4 in group B-S, 3.2 in group B-L, 1.0 in group C-S, and 2.9 in group C-L. Compared with group R-O, the torques to failure of groups B-S and C-S were significantly lower, whereas only group C-S was significantly weaker than group R-P (P < .01). Comparing short- and long-stem bypasses of different fracture heights, we found that only group C-L showed a significantly higher resistance to torque (P < .01). A short-stem bypass with a BI of 1.7 was sufficient for primary stability tested by torque to failure in this biomechanical setting. For smaller BIs, a long-stem prosthesis should be considered. Basic science study, biomechanics.


Prevalence and characteristics of disinhibition during bronchoscopy with midazolam.
Disinhibition is sometimes experienced during bronchoscopy with sedation. However, data on disinhibition during bronchoscopy are scarce. We examined the prevalence and characteristics of disinhibition during bronchoscopy with midazolam. This retrospective study analyzed consecutive patients who underwent bronchoscopy between November 2019 and December 2020. The severity of disinhibition was defined as follows: mild, disinhibition sometimes requiring restraints by assistants; moderate, disinhibition always requiring restraints by assistants; and severe, disinhibition requiring antagonization of sedation by flumazenil to continue bronchoscopy. Among 251 eligible patients who were sedated using midazolam, 36 (14.3%; 95% confidence interval [CI], 10.5%-19.2%), 42 (16.7%; 95% CI, 12.6%-21.8%), and 7 (2.8%; 95% CI, 1.4%-5.6%) experienced mild, moderate, and severe disinhibition, respectively. Depression (odds ratio [OR] 2.77; 95% CI, 1.20-6.41), endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) (OR 10.23; 95% CI, 1.02-103.01, referred to brushing/bronchial washing/observation), and increased administration of midazolam (OR 1.20; 95% CI, 1.02-1.42, per 1-mg increase) were independently associated with moderate-to-severe disinhibition. Patients experiencing moderate disinhibition reported significantly better scores for discomfort during bronchoscopy. Besides the maximum systolic and diastolic blood pressures during bronchoscopy, the changes in hemodynamic and respiratory statuses during bronchoscopy or complications did not significantly differ between patients experiencing moderate-to-severe disinhibition and those experiencing none-to-mild disinhibition. Moderate-to-severe disinhibition occurred in 19.5% of patients during bronchoscopy with midazolam. We should focus on disinhibition when patients have depression or are planning to undergo EBUS-TBNA, and sparing the administration of midazolam might reduce the occurrence of disinhibition. UMIN000038571.


Changes in lactate and other ions in plaque and saliva after a fluoride rinse and subsequent sucrose administration.
The purpose of this study was to examine plaque and saliva composition after a fluoride rinse and subsequent sucrose application. Fifteen subjects accumulated plaque for 48 h, and then rinsed with a fluoride rinse based on 228 microg/g (ppm) Na2SiF6 and some received no rinse. After 60 min, upper and lower buccal molar plaque samples and 1-min saliva samples were collected. The subjects then rinsed with 10% g/g sucrose solution, and 7 and 15 min later, a second and a third set of samples were collected. Plaque fluid and clarified saliva were then recovered from these samples by centrifugation, and the remaining plaque acid extracted. The plaque fluid, centrifuged saliva, and plaque extract samples were then analyzed using micro techniques for pH, free calcium, phosphate, organic acids (plaque fluid and saliva only) and fluoride. Considering both the fluoride rinse and no-rinse groups, the most notable compositional changes in saliva 7 min after the sucrose rinse were pH -0.40 unit, free calcium 0.42 mM, lactate 5.2 mM, phosphate -1.3 mM, and fluoride 2.8 microM; while in plaque fluid, the corresponding changes were pH -1.59 unit, free calcium 1.5 mM, lactate 35 mM, phosphate -1.6 mM and fluoride -26 microM. After sucrose rinsing, undersaturation was found with respect to dicalcium phosphate dihydrate in saliva and plaque fluid and with respect to tooth enamel in some plaque fluid samples. Plaque fluid composition appeared to be strongly influenced by salivary clearance, diffusive loss of ions into the water phase of the rinse, and lower jaw pooling of the sucrose and fluoride components of the rinses. After the experimental rinse, the fluoride concentration in plaque fluid [86 +/- 22 mM (upper molar site), 162 +/- 150 mM (lower molar site)], saliva (26 +/- 18 mM), and whole plaque [99 +/- 97 microg/g (upper molar site), 197 +/- 412 microg/g (lower molar site)] was comparable to the values in previous studies using this rinse. These very high plaque fluid fluoride concentrations, compared with the 'no-rinse' samples, induced an approximately 0.3-unit increase in the plaque fluid pH 7 min after the sucrose rinse, a small decrease (approximately 20%) in lactate production and a modest increase in enamel saturation. Although these changes were all statistically significant, no correlation was found between the decrease in lactate concentration and plaque fluid fluoride, pH or whole plaque fluoride.


Four-Component Relativistic Density Functional Theory Calculations of EPR g- and Hyperfine-Coupling Tensors Using Hybrid Functionals: Validation on Transition-Metal Complexes with Large Tensor Anisotropies and Higher-Order Spin-Orbit Effects.
The four-component matrix Dirac-Kohn-Sham (mDKS) implementation of EPR g- and hyperfine A-tensor calculations within a restricted kinetic balance framework in the ReSpect code has been extended to hybrid functionals. The methodology is validated for an extended set of small 4d(1) and 5d(1) [MEXn](q) systems, and for a series of larger Ir(II) and Pt(III) d(7) complexes (S = 1/2) with particularly large g-tensor anisotropies. Different density functionals (PBE, BP86, B3LYP-xHF, PBE0-xHF) with variable exact-exchange admixture x (ranging from 0% to 50%) have been evaluated, and the influence of structure and basis set has been examined. Notably, hybrid functionals with an exact-exchange admixture of about 40% provide the best agreement with experiment and clearly outperform the generalized-gradient approximation (GGA) functionals, in particular for the hyperfine couplings. Comparison with computations at the one-component second-order perturbational level within the Douglas-Kroll-Hess framework (1c-DKH), and a scaling of the speed of light at the four-component mDKS level, provide insight into the importance of higher-order relativistic effects for both properties. In the more extreme cases of some iridium(II) and platinum(III) complexes, the widely used leading-order perturbational treatment of SO effects in EPR calculations fails to reproduce not only the magnitude but also the sign of certain g-shift components (with the contribution of higher-order SO effects amounting to several hundreds of ppt in 5d complexes). The four-component hybrid mDKS calculations perform very well, giving overall good agreement with the experimental data.


Percutaneous penile venoablation for treatment of impotence.
Percutaneous penile venoablation with sclerosing agents and coils was performed in 58 patients. Of 104 ablations attempted, 44 of 51 (86%) transpenile, and 46 of 53 (86.7%) retrograde interventions were technically successful. In 40 patients (69%) erectile function improved (intercourse was possible in 21 without additional measures). Initially improved erectile function deteriorated in 24 patients during follow-up (in an average of 6 months); in 15 of these, intercourse was still possible with injection of vasoactive drugs. In 7 patients, repeat venoablation improved erectile function again. Hot contrast medium was slightly superior to sodium morrhuate as a sclerosing agent. In successfully treated patients, the average decrease in venous leakage was 30 ml/min vs 13 ml/min in treatment failures, with considerable overlap between both groups. Only minor complications were observed. We conclude that percutaneous penile venoablation is technically feasible, and safe. Whether the procedure will have a definite role in the treatment of venogenic impotence, however, still has to be determined.


Kashin-Beck disease and Sayiwak disease in China: prevalence and a comparison of the clinical manifestations, familial aggregation, and heritability.
To compare the prevalence, the clinical manifestations, familial aggregation and heritability of Kashin-Beck disease (KBD) and Sayiwak disease (SD) in China. 10,823 people from 1361 families in 14 villages in Linyou County, Shaanxi Province, were examined for KBD, and 2264 people from 552 families in Sayiwak village, Kashi city, Xinjiang, were examined for SD. The investigation included documentation of individual information and clinical manifestations. Patients were subject to radiographic imaging of the right hand. t-Tests and chi-square tests were used to examine correlations of the diseases with age and gender in each of the two groups. Analysis of familial aggregation was conducted with the chi-square distribution analysis of goodness of fit using the SAS8.0 program. The Li-Mantel-Gart method was employed for the segregation analysis. The Falconer regression method11 was employed to estimate heritability (h²). The prevalence of KBD in Linyou County was 10.90%, and of SD in Sayiwak village was 0.57%. Of the 21 clinical signs examined, KBD cases exhibited 19 signs (90.48%) and SD cases exhibited 18 signs (85.71%), which indicate similarities between the two diseases. However, different clinical signs were evident between the KBD and SD cases, with different impairment rates among joints of limbs in KBD and similar rates in SD. A comparison of radiological features of limb arthropathy between the two diseases showed differences in several characteristics between the two diseases. In addition, measurements of stature and sitting height showed significant differences in bone development between the two diseases. For KBD cases, the values of h² in the first-degree and the second-degree relatives were 41.76% and 37.20% (P<0.05). The CI of h² was 31.17-52.38 and 19.86-54.55, with a segregation ratio of P=0.12, SE(P)=0.014, 95%CI 0.09-0.15, less than 0.25(χ²=42.36, df=1, P<0.001). For SD cases, the values of h² were 155.61%, 273.63% and 236.83%. The 95% CIs of h² were 133.20-178.12, 229.83-317.42 and 145.83-327.81, respectively, with a segregation ratio of P=0.34, SE(P)=0.059, and CIs between 0.22 and 0.45(χ²=4.9817, df=1, P>0.05). The results indicate both similarities and differences in the clinical manifestations of KBD and SD. However, environmental factors appear to play a major role in KBD, while hereditability is a major factor in SD.


Surgical treatment for pelvic bone metastatic disease from renal cell carcinoma.
Pelvic bone metastases from renal cell carcinoma (RCC) are challenging due to their destructive pattern, poor response to radiotherapy and hypervascularity. The purpose of our study was to review a series of patients undergoing surgical treatment with the aim to investigate: 1) survival; 2) local disease control; and 3) complications. A series of 16 patients was reviewed. A curettage procedure was performed on 12 patients. In eight the lesion involved the acetabulum; a cemented hip arthroplasty with cage was performed in seven; a flail hip in one. Four patients received a resection; in two cases with acetabular involvement, reconstruction was performed with a custom-made prosthesis and with an allograft and prosthesis. Disease-specific survival accounted for 70% at 3 years and 41% at 5 years. Only one local tumor progression after curettage occurred. Revision surgery (flail hip) was necessary for deep infection of the custom-made prosthesis. A prolonged survival in patients affected by bone metastatic disease from RCC can justify also major surgical procedures. Due to a low local progression rate after intralesional procedures, curettage, cement and a total hip arthroplasty with cage, when feasible, should be considered as an alternative to more demanding surgeries like resections and reconstructions. Level 4.


Inhibitory cotransmission or after-hyperpolarizing potentials can regulate firing in recurrent networks with excitatory metabotropic transmission.
Recurrent networks of neurons communicating via excitatory connections are common in the nervous system. In the absence of mechanisms to control firing (collectively termed negative feedback), these networks are likely to be bistable and unable to meaningfully encode input signals. In most recurrent circuits, negative feedback is provided by a specialized subpopulation of interneurons, but such neurons are absent from some systems, which therefore require other forms of negative feedback. One such circuit is found within the enteric nervous system of the intestine, where AH/Dogiel type II neurons are interconnected via excitatory synapses acting through metabotropic receptors to produce slow excitatory postsynaptic potentials (slow EPSPs). Negative feedback in this recurrent network may come from either inhibitory postsynaptic potentials arising from the terminals that produce slow EPSPs or from the after hyperpolarizing potentials (AHPs) characteristic of these neurons. We have examined these possibilities using mathematical analysis, based on the Wilson-Cowan model, and computer simulations. Analysis of steady states showed that, under appropriate conditions, both types of negative feedback can provide robust regulation of firing allowing the networks to encode input signals. Numerical simulations were performed using large, anatomically realistic networks with realistic models for metabotropic transmission and suppression of the AHP. In the presence of constant exogenous input, parameters controlling aspects of synaptic events were varied, confirming the analytical results for static stimuli. The simulated networks also responded to time varying inputs in a manner consistent with known physiology. In addition, simulation revealed that neurons in networks with inhibitory contransmission fired in erratic bursts, a phenomenon observed in neurons in unparalysed tissue. Thus, either inhibitory contransmission or AHPs, or both, can allow recurrent networks of AH/Dogiel type II neurons to encode ongoing inputs in a biologically useful way. These neurons appear to be intrinsic primary afferent neurons (IPANs), which implies that the IPANs in a region act in a coordinated fashion.


Effects of bradykinin on cell volume and intracellular pH in NIH 3T3 fibroblasts expressing the ras oncogene.
BCECF fluorescence has been applied to determine intracellular pH (pHi) in NIH 3T3 fibroblasts expressing the Ha-ras oncogene (+ras) and otherwise identical cells not expressing the oncogene (-ras). In +ras cells, pHi is significantly more alkaline (6.79 +/- 0.03 n = 12) than in -ras cells (6.64 +/- 0.02, n = 8). Bradykinin (100 nmol/l) leads to intracellular alkalinization in both +ras (to 6.96 +/- 0.04, n = 12) and -ras cells (to 6.85 +/- 0.02, n = 8). The effect of bradykinin is completely abolished in the presence of dimethylamiloride (100 mumol/l), which does not modify pHi in the absence of bradykinin. Similar to bradykinin, cell shrinkage by addition of 15 mmol/l NaCl to the extracellular fluid leads to intracellular alkalinization (by 0.08 +/- 0.01, n = 15). Cell volume is significantly greater in +ras cells (2.7 +/- 0.4 pl, n = 15) than in -ras cells (2.2 +/- 0.4 pl, n = 15). Bradykinin leads to cell shrinkage in both +ras cells (by 7 +/- 1%, n = 17) and -ras cells (by 5 +/- 1%, n = 15). The effect of bradykinin on cell volume can be reversed by the reduction of extracellular NaCl concentration by 15 mmol/l NaCl in +ras cells and by 7 mmol/l NaCl in -ras cells. This maneuver completely abolishes (in -ras cells) or blunts (in +ras cells) the alkalinizing effect of bradykinin. In conclusion, +ras cells are more alkaline than -ras cells. Bradykinin leads to further intracellular alkalinization by activation of the Na+/H(+)-exchanger, at least in part secondary to hormone-induced cell shrinkage.


Recurrent ventricular septal defect following closure CorMatrix: A case report.
Ventricular septal ruptures are an uncommon complication following acute myocardial infarction. Operative repair, utilizing a patch for closure of the defect, is the primary treatment modality to achieve hemodynamic stability. The use of an extracellular matrix derived from small intestinal submucosa as a scaffold for tissue repair is becoming increasingly common. Here, we present the case of a 58-year-old female found to have a ventricular septal rupture and posterior left ventricular aneurysm following late presentation after a myocardial infarction that required operative repair with a CorMatrix patch. Upon readmission for dyspnea and poor exercise tolerance several months later, the patch was subsequently found to have near-completely reabsorbed. There is a paucity of long-term outcomes data following the use of CorMatrix for septal defects, with rare reports of such reabsorption. Further study is required to identify the incidence and implications of such findings.


[Gastric calcifying fibrous pseudotumor, a rare mesenchymal tumor of the stomach].
We report a case of calcifying fibrous pseudotumor fortuitously discovered on abdominal CT-scan in a 50 year-old woman presenting with metrorragia. CT-scan showed a 2 cm hypodense nodular tumor of the gastric wall with granular calcifications. Upper endoscopy showed a nodular tumor along the lesser curvature of the gastric body, covered by normal gastric mucosa and biopsies were negative. A diagnosis of gastric stromal tumor was suspected and a partial gastrectomy was performed. On gross examination surgical specimen showed a firm, whitish nodular tumor measuring 2 cm in diameter. On microscopic examination the tumor was composed of whorls of dense hyalinized collagen bundles with a few fibroblasts. There were psammomatous calcifications and nodular aggregates of mononuclear inflammatory cells. Immunohistochemically, spindle cells stained for vimentin. They did not stain for smooth muscle actin, desmin, CD34, CD117, S100 protein, Ki-67 and ALK-1. Surgical resection of the tumor was complete. Patient has no evidence of disease with a follow-up of 12 months. Only seven cases of gastric calcifiying fibrous pseudotumors have been reported in the literature. These benign tumors are usually incidentally discovered. They have characteristic imaging and microscopic features and appear as a distinct clinicopathologic entity different from stromal tumors.


Lumped kinetics and acute toxicity of intermediates in the ozonation of phenol in saline media.
This work investigates the feasibility of ozonation for destroying phenol and removing organic matter in saline media. The reaction lumped kinetics was followed using the GLKM (General Lumped Kinetic Model). The main intermediate compounds were: catechol, hydroquinone, 4,4'-dihydroxybiphenyl, and 4-bromophenol. It could be noted no significant differences in phenol degradation, mineralization rates, and toxicity removal up to 2 g L(-1) of salt. So, ozonation appears to be a technology that can be used in low salinity media, which is characteristic of waters destined to reuse and recycling programs inside industries.


Thermodynamics of the various forms of the dodecamer d(ATTACCGGTAAT) and of its constituent hexamers from proton nmr chemical shifts and UV melting curves: three-state and four-state thermodynamic models.
Chemical shifts of base and H1' protons of the single-stranded hexamers d(ATTACC) and d(GGTAAT), of the 1:1 mixtures of these complementary hexamers, and of the self-complementary dodecamer d(ATTACCGGTAAT) were measured at various temperatures in aqueous solution. Four different sample concentrations were used in the case of the dodecamer and of the mixture of the complementary hexamers; the individual hexamers were measured at two different DNA concentrations. Absorbance temperature profiles at five different NaCl concentrations were measured for the dodecamer in order to quantify the effect of the ionic strength on the duplex formation. Under suitable conditions of nucleotide concentration, temperature, and ionic strength, the dodecamer adopts either a B-DNA duplex or a hairpin-loop structure. Chemical shift vs temperature profiles, constructed for all samples, were used to obtain thermodynamic parameters either for the various stacking interactions in the single strands or for the duplex or the hairpin-loop formation. In the analysis of the duplex formation of the hexamers, a two-state approach appeared too simple, because systematic deviations were revealed. Therefore, a new three-state model (DUPSTAK) was developed. In order to investigate the magnitude of error arising from the use of the two-state approach in cases where the DUPSTAK model appears more appropriate, a series of test calculations was made. The magnitude of error in the enthalpy and in the entropy of duplex melting is found to depend linearly upon the actual melting temperature and not upon the individual delta Hd degrees and delta Sd degrees values. Thermodynamic analysis of the chemical shift vs temperature profiles in D2O solution (no added salt) yields an average Tmd value of 341 K (1M DNA) and delta Hd degrees of - 121 kJ.mol-1 for the dimer/random-coil transition of the hexamer duplex d(ATTACC).d(GGTAAT). For the duplex in equilibrium random-coil transition of the 12-mer d(ATTACCGGTAAT) an average Tmd value of 336 K (1M DNA) and delta Hd degrees of -372 kJ.mol-1 are found. The hairpin/random-coil transition of d(ATTACCGGTAAT) is characterized by a rather large delta Hh degrees value, -130 kJ.mol-1, and an average Tmh value of 304 K.


An unusual presentation of Brown-Sequard syndrome.
We report the case of a man with Brown-Sequard syndrome following a fall in which he sustained fractures of the thoracic and lumbosacral spine. Despite characteristic neurological findings, the diagnosis was delayed due to the absence of history of penetrating spinal trauma and incorrect attribution of unilateral-like weakness and numbness to lumbosacral trauma. A directed history and examination revealed that the patient was stabbed in the back with a penknife while leaving a bus and stepped down onto a paralyzed leg, which collapsed beneath him. The patient was given an antibiotic, underwent a negative peritoneal lavage, and had myelography and nerve conduction velocity confirmatory for Brown-Sequard syndrome. With rehabilitation he became ambulatory with a cane one month after the stabbing.


Mechanical properties of the rotator cuff: response to cyclic loading at varying abduction angles.
The rotator cuff is loaded under static as well as dynamic conditions. Whilst the static properties of the rotator cuff muscle-tendon junctions have been reported, the dynamic mechanical behaviour has not. This study reports the dynamic mechanical properties with varying abduction angles in a human cadaver rotator cuff. No significant effect was found with varying the angle of testing or in the presence of a tear in the tendon. The supraspinatus was found to be the stiffest of the rotator cuff tendons followed by the subscapularis and infraspinatus.


Pipkin type III femoral head fracture: which treatment strategy can be recommended?
Pipkin type III femoral head fractures are relatively rare injuries. Few studies have explored and described the treatment and outcomes of Pipkin type III femoral head fractures. The purpose of this study was to evaluate the efficacy of open reduction and internal fixation (ORIF) in treating Pipkin type III femoral head fractures. We retrospectively reviewed 12 patients with Pipkin type III femoral head fractures who underwent ORIF from July 2010 and January 2018. The complications and reoperations were recorded. The visual analog scale (VAS) pain score, Harris hip score (HHS), Thompson-Epstein criteria, and SF-12 score [including the physical component summary (PCS) and the mental component summary (MCS)] were used for functional assessment. Among the 12 patients, ten were males and two were females, with a mean age of 34.2 ± 11.9 years. The median follow-up time was 6 years (range 4-8 years). Five patients (42%) developed osteonecrosis of the femoral head, and one patient (8%) developed nonunion. These six patients (50%) underwent total hip arthroplasty (THA). One patient (8%) developed heterotopic ossification and underwent ectopic bone excision; this patient also presented with post-traumatic arthritis. The mean final VAS pain score and HHS were 4.1 ± 3.1 points and 62.8 ± 24.4 points, respectively. According to the Thompson-Epstein criteria, there was one patient (8%) with excellent, four patients (33%) with good, one patient (8%) with fair, and six patients (50%) with poor outcomes. The PCS score and MCS score were 41.7 ± 34.7 points and 63.2 ± 14.5 points, respectively. Limited by the high incidence of osteonecrosis of the femoral head, it is difficult to achieve satisfactory functional outcomes when treating Pipkin type III femoral head fractures using ORIF, and a primary THA may be considered. However, for younger patients, considering the survivorship of prosthesis, ORIF may be recommended with the proviso that the patient is fully informed of the high complication rate associated with this procedure. IV.


Antioxidant status of rats receiving lycopene in different doses.
Oral treatment with lycopene (per os) in doses of 10 or 50 mg/kg for 2 weeks led to accumulation of lycopene in the liver, liver microsomes, and blood plasma, increased total plasma antioxidant activity, inhibited LPO in the liver, and decreased solubilization of lysosomal enzymes. Lycopene had no effect on ex vivo resistance of liver microsomes to LPO and activities of antioxidant enzymes in the liver.


A comparison of decision-making processes for conventional and complementary medicine in cancer patients.
To understand the decision-making processes involved in conventional medicine and complementary medicine cancer treatment, and the role that information plays, a cross-sectional survey of cancer patients was carried out at 2 Australian cancer centres. Data was obtained from 75 cancer patients. When asked about complementary medicine use, 36 (48%) patients reported using some form of complementary medicine in conjunction with their conventional treatment. In terms of decision-making, our findings suggest that although cancer patients appear to be at different stages of decision-making for complementary medicine compared with conventional medicine, there was no difference between the decision-making process for cancer patients when it comes to utilising conventional medicine or complementary medicine: patients reported comparable high decisional self-efficacy and low decisional conflict for both areas of medicine.


Performance of brief ICF-sleep disorders and obesity core set in obstructive sleep apnea patients.
Clinical questionnaires are mainly applied as screening tools for identification of the Obstructive sleep apnea (OSA) patients. Little attention has been paid to assess the body functions and health status of the patients. International Classification of Functioning, Disability and Health (ICF) was designed for better understanding and describing functioning and disability of patients. This study adopted the Brief ICF-Sleep Disorders and Obesity Core Set to evaluate the impairment of functioning and health status of OSA patients. Five hundred ninety-two participants were enrolled in this cross-sectional study. Data were collected using Brief ICF-Sleep Disorders and Obesity Core Set Polysomnography was performed and basic characteristics of the patients were recorded. The scores for the component Body Functions and Code b130, b134, b140, b440, b530, s330, d160, d240, d450 of the two core sets were significantly different among the patients divided by apnea-hypopnea index (AHI) or oxygen saturation (SaO2) nadir, but the frequency of code s330, d160, d240, d450 was low. The Body Functions component of the both sets were closely related to neck circumference (NC), body mass index (BMI), apnea-hypopnea index (AHI) of the OSA patients. Body Functions of the Brief ICF-Sleep Disorders performed better with a threshold of 4 with sensitivity, specificity and area under the receiver operating characteristic curve (AUC) as 0.62, 0.74, 0.68(AHI ≥ 5), 0.69, 0.63, 0.66 (AHI ≥ 15), 0.75, 0.56, 0.66 (AHI ≥ 30), 0.56, 0.70, 0.63 (SaO2 nadir≤90%), 0.67, 0.66, 0.66 (SaO2 nadir<85%), 0.71, 0.59, 0.65 (SaO2 nadir<80%), separately. The Body Functions component of both two sets could be an evaluation tool of impairment of body functions for OSA patients. The Brief ICF-Sleep Disorders Body Functions component performed better with a threshold of 4 and might provide a new insight for physicians to assess OSA patients.


Glucose deprivation results in the induction of glucose-regulated proteins and domes in MDCK monolayers in hormonally defined serum-free medium.
Madin-Darby canine kidney (MDCK) cells spontaneously form dome-like structures in vitro, a phenomenon which has been proposed to be indicative of cellular differentiation. This study indicates the existence of a correlation between the induction of domes and of glucose-regulated proteins during glucose starvation. When MDCK monolayers were glucose deprived, domes appeared very rapidly. After only 3 h of glucose deprivation domes appeared in 69% of the microscope fields. The level of expression of glucose-regulated proteins (grps) as well as domes was examined over a 6-h time interval of glucose deprivation. Both grp 76 and 97 were induced over this time interval, with grp 76 being the more readily detectable. The level of induction of grp 76 as a function of time was quantitated by means of densitometry measurements. The induction of domes was examined in parallel with the induction of grp 76. The results indicated that the induction of grp 76 and domes occurs with a similar time course. The effect of glucose deprivation on the initial rate of ouabain-sensitive Rb+ uptake was also examined. Within the first 4 h of glucose deprivation, the initial rate of ouabain-sensitive Rb+ uptake did not differ significantly in glucose-deprived and control MDCK monolayers. These observations indicate that unlike the case with other methods of dome induction (e.g., treatment with either prostaglandin E1 (PGE1) or hexamethylene bis-acetamide (HMBA] glucose deprivation does not affect the Na+K+ ATPase activity of MDCK monolayers. These observations suggest that PGE1, HMBA, and glucose deprivation affect dome formation in MDCK monolayers by means of distinct mechanisms.


Duplication is more common among laterally transferred genes than among indigenous genes.
Recent developments in the understanding of paralogous evolution have prompted a focus not only on obviously advantageous genes, but also on genes that can be considered to have a weak or sporadic impact on the survival of the organism. Here we examine the duplicative behavior of a category of genes that can be considered to be mostly transient in the genome, namely laterally transferred genes. Using both a compositional method and a gene-tree approach, we identify a number of proposed laterally transferred genes and study their nucleotide composition and frequency of duplication. It is found that duplications are significantly overrepresented among potential laterally transferred genes compared to the indigenous ones. Furthermore, the GC3 distribution of potential laterally transferred genes was found to be largely uniform in some genomes, suggesting an import from a broad range of donors. The results are discussed not in a context of strongly optimized established genes, but rather of genes with weak or ancillary functions. The importance of duplication may therefore depend on the variability and availability of weak genes for which novel functions may be discovered. Therefore, lateral transfer may accelerate the evolutionary process of duplication by bringing foreign genes that have mainly weak or no function into the genome.


Strain rate and mineral content in fracture models of bone.
I reanalysed data from my previous work to determine the extent to which a model for the loading-rate dependence of the fracture of cortical bone, put forward by Carter and Caler, fits this independently derived data set. In particular, the extent to which the generality of the model is vitiated by its ignoring the effect of mineralisation on strength was tested. The model was rather strongly corroborated. In addition, the reanalysed data show that yield strain is strongly strain-rate dependent, but that Young's modulus is rather unvarying over physiological strain rates. The implications of this for hypotheses concerning fracture of bone are discussed.


Determination of post-culture processing with carbohydrates by MALDI-MS and TMS derivatization GC-MS.
Biological materials generally require stabilization to retain activity or viability in a dry form. A number of industrial products, such as vaccines, probiotics and biopesticides have been produced as dry preparations. The same methods and materials used for stabilizing commercial microbial products may be applicable to preserving biothreat pathogens in a dry form. This is a likely step that may be encountered when looking at samples from terrorism attempts since only spores, such as those from Bacillus anthracis, are inherently stable when dried. The stabilizers for microbial preparations generally include one or more small carbohydrates. Different formulations have been reported for different industrial products and are often determined empirically. However sugar alcohols (mannitol and sorbitol) and disaccharides (lactose, sucrose and trehalose) are the common constituents of these formulations. We have developed an analytical method for sample preparation and detection of these simple carbohydrates using two complementary analytical tools, MALDI-MS and GC-MS. The native carbohydrates and other constituents of the formulation are detected by MALDI-MS as a screening tool. A longer and more detailed analysis is then used to specifically identify the carbohydrates by derivatization and GC-MS detection. Both techniques were tested against ten different types of stabilization recipes with Yersinia pestis cell mass cultured on different media types used as the biological component. A number of additional components were included in these formulations including proteins and peptides from serum or milk, polymers (e.g. poly vinyl pyrrolidone - PVP) and detergents (e.g. Tween). The combined method was characterized to determine several figures of merit. The accuracy of the method was 98% for MALDI-MS and 100% for GC-MS. The repeatability for detection of carbohydrates by MALDI-MS was determined to be 96%. The repeatability of compound identification by GC-MS was determined by monitoring variation in retention time, which is vital for identification of isomeric carbohydrates. The figures of merit illustrate an effective and accurate method for mono and disaccharide detection independent of formulation. This meets our primary goal for method development as small carbohydrates are among the most common stabilizers employed.


Onset of action of formoterol versus salmeterol via dry powder inhalers in moderate chronic obstructive pulmonary disease: a randomized, placebo-controlled, double-blind, crossover study.
Bronchodilator therapy is central to the symptomatic management of chronic obstructive pulmonary disease (COPD), and treatment with short-acting bronchodilators is recommended in patients with mild COPD. This study aimed to evaluate the onset of effect of single-dose formoterol 9 μg versus single-dose salmeterol 50 μg in patients with moderate COPD. In this multicentre, double-blind, double-dummy, placebo-controlled, three-way single-dose crossover study, patients ≥40 years of age with moderate COPD were randomized to single-dose formoterol 9 μg via Turbuhaler® plus placebo via Diskus®, single-dose salmeterol 50 μg via Diskus® plus placebo via Turbuhaler® or placebo via Turbuhaler® and Diskus® (washout period 2-7 days). Terbutaline 0.5 mg/actuation via Turbuhaler® was used as reliever medication throughout. The primary endpoint was forced expiratory volume in 1 second (FEV₁) at 5 minutes post-dose. Secondary endpoints included proportion of patients achieving ≥12% increase in FEV₁ at 5 minutes post-dose. 109 patients were randomized, and 108 completed the study. The increase in FEV₁ 5 minutes post-dose versus pre-dose was 7.2% for formoterol, 4.1% for salmeterol and 0.7% for placebo, and significantly greater for formoterol versus salmeterol (ratio of treatment effects: 1.030; 95% CI 1.008, 1.052; p = 0.009), for formoterol versus placebo (1.064, 95% CI 1.041, 1.087; p < 0.001) and for salmeterol versus placebo (1.033, 95% CI 1.011, 1.056; p = 0.003). The proportions of patients with ≥12% increase in FEV₁ 5 minutes post-dose were 23.1%, 9.2% and 6.4% for formoterol, salmeterol and placebo, respectively; this was statistically significantly larger after formoterol than salmeterol (p = 0.008) or placebo (p < 0.001). All treatments were well tolerated. In COPD patients, formoterol 9 μg has an onset of bronchodilatory effect that is more rapid than salmeterol 50 μg based on FEV₁ at 5 minutes post-dose. Clinicaltrials.gov identifier: NCT01048333; AstraZeneca study code: D5127C00001.


Blockade by haloperidol of the increase in tryptophan hydroxylase activity induced by incubation of slices of brain stem with dibutyryl cyclic AMP.
Pretreatment of rat brain stem slices with dibutyryl cyclic AMP, caffeine, theophylline and 3-isobutyl-1-methylxanthine increased the activity of tryptophan hydroxylase in supernatant preparations of enzyme made from the slices. This effect does not appear to be mediated by a cyclic AMP sensitive mechanism since it was not reproduced by exposure of the slices to 8-bromo cyclic AMP, to papaverine, a nonxanthine phosphodiesterase inhibitor, or to other treatments known to raise tissue cyclic AMP levels. The ability of haloperidol to block this increase in enzyme activity is consistent with a role for calmodulin and calcium as mediators of the enzyme activation, particularly when this observation is considered in conjunction with the evidence that supernatant preparations of this enzyme was activated under phosphorylating conditions by a calcium-calmodulin dependent process [5]. Nevertheless, in view of the high concentrations of haloperidol employed in the present experiments, the possibility that this drug may produce its effects in the brain stem slices through some other action, unrelated to its ability to bind to calmodulin, should be kept in mind.


Diterpene resin acid biosynthesis in loblolly pine (Pinus taeda): functional characterization of abietadiene/levopimaradiene synthase (PtTPS-LAS) cDNA and subcellular targeting of PtTPS-LAS and abietadienol/abietadienal oxidase (PtAO, CYP720B1).
Diterpene resin acids are prominent defense compounds against insect pests and pathogens in conifers. Biochemical and molecular analyses in grand fir (Abies grandis), Norway spruce (Picea abies), and loblolly pine (Pinus taeda) have identified two classes of genes and enzymes that generate much of the structural diversity of terpenoid defense compounds: The terpenoid synthases (TPS) and cytochrome P450 monooxgenases (P450). Using a single substrate, geranylgeranyl diphosphate, families of single-product and multi-product diterpene synthases generate an array of cyclic diterpene olefins. These diterpenes are converted to diterpene resin acids by activity of one or more P450 enzymes. A few conifer diterpene synthases have previously been cloned and characterized in grand fir and in Norway spruce. We have also previously shown that the loblolly pine P450 abietadienol/abietadienal oxidase (PtAO) catalyzes multiple oxidations of several diterpene alcohols and aldehydes. Conifer diterpene synthases are thought to function in plastids while P450s can also be localized to plastids or to the endoplasmic reticulum (ER). Here, we show that a loblolly pine cDNA (PtTPS-LAS) encodes a typical multi-product conifer diterpene synthase that forms levopimaradiene, abietadiene, palustradiene, and neoabietadiene similar to the grand fir abietadiene synthase and Norway spruce levopimaradiene/abietadiene synthase. Subcellular targeting of PtTPS-LAS and PtAO to plastids and ER, respectively, was shown with green fluorescent fusion protein expression in tobacco cells. These data suggest that enzymes for conifer diterpene resin acid biosynthesis are localized to at least two different subcellular compartments, plastids and ER, requiring efficient transport of intermediates and secretion of diterpene resin acids into the extracelluar space.


Initial experiences of tissue harmonic imaging in the diagnosis of fetal cardiac tumors.
The usefulness of tissue harmonic imaging in prenatal diagnosis is illustrated in two fetuses with a cardiac tumor. Tissue harmonic imaging provided more informative images than conventional B-mode imaging, enabling the detection of the site of attachment of the cardiac tumor and estimation of intracardiac blood flow. The advantages of tissue harmonic imaging over conventional B-mode imaging in prenatal diagnosis are discussed.


Factors influencing ester synthesis catalysed in aqueous media by the lipase from Candida deformans (Zach) Langeron and Guerra.
The purified lipase from Candida deformans was shown to catalyse ester production in aqueous media by esterification of free fatty acids but not by alcoholysis. As the enzyme also catalysed ester hydrolysis, the influence of various physico-chemical factors on ester hydrolysis and synthesis was studied and compared. Substrate specificities were also studied. Both activities had the same pH and temperature optima, and did not require a metal cofactor. Tyrosine appeared to be one of the amino acids of the enzyme required by both catalytic activities, whereas serine-reactive reagents inhibited synthesis only. The highest synthesis and hydrolysis activities were obtained with mono-, di- and tri-unsaturated fatty acids containing a [cis] delta-9 unsaturation and 16 to 18 carbon atoms. Only esters of primary alcohols were hydrolysed and synthesised. For alcohols ranging from methanol to butanol, synthesis activity increased with the length of the alcohol whereas hydrolysis activity of the corresponding esters decreased. The presence of alcohol inhibited hydrolysis. An optimum concentration was found for each primary alcohol at which ester synthesis was maximum and hydrolysis was low.


Involvement of μ-opioid receptors in antinociceptive action of botulinum toxin type A.
Botulinum toxin A (BTX-A) is approved for treatment of chronic migraine and has been investigated in various other painful conditions. Recent evidence demonstrated retrograde axonal transport and suggested the involvement of CNS in antinociceptive effect of BTX-A. However, the mechanism of BTX-A central antinociceptive action is unknown. In this study we investigated the potential role of opioid receptors in BTX-A's antinociceptive activity. In formalin-induced inflammatory pain we assessed the effect of opioid antagonists on antinociceptive activity of BTX-A. Naltrexone was injected subcutaneously (0.02-2 mg/kg) or intrathecally (0.07 μg/10 μl-350 μg/10 μl), while selective μ-antagonist naloxonazine was administered intraperitoneally (5 mg/kg) prior to nociceptive testing. The influence of naltrexone (2 mg/kg s.c.) on BTX-A antinociceptive activity was examined additionally in an experimental neuropathy induced by partial sciatic nerve transection. To investigate the effects of naltrexone and BTX-A on neuronal activation in spinal cord, c-Fos expression was immunohistochemically examined in a model of formalin-induced pain. Antinociceptive effects of BTX-A in formalin and sciatic nerve transection-induced pain were prevented by non-selective opioid antagonist naltrexone. Similarly, BTX-A-induced pain reduction was abolished by low dose of intrathecal naltrexone and by selective μ-antagonist naloxonazine. BTX-A-induced decrease in dorsal horn c-Fos expression was prevented by naltrexone. Prevention of BTX-A effects on pain and c-Fos expression by opioid antagonists suggest that the central antinociceptive action of BTX-A might be associated with the activity of endogenous opioid system (involving μ-opioid receptor). These results provide first insights into the mechanism of BTX-A's central antinociceptive activity.


Association between CLOCK 3111 T/C polymorphism with ghrelin, GLP-1, food timing, sleep and chronotype in overweight and obese Iranian adults.
Circadian Locomotor Output Cycles Kaput (CLOCK), an essential element of the positive regulatory arm in the human biological clock, is involved in metabolic regulation. The aim was to investigate the behavioral (sleep duration, food timing, dietary intake, appetite and chronobiologic characteristics) and hormonal (plasma ghrelin and Glucagon-like peptide-1 concentrations) factors that could explain the previously reported association between the CLOCK 3111 T/C SNP and obesity. This cross-sectional study included 403 subjects, overweight and/or obesity, aged 20- 50 years from Iran. The CLOCK rs1801260 data were measured by the PCR-RFLP method. Dietary intake, food timing, sleep duration, appetite and Chrono-type were assessed using validated questionnaires. Ghrelin and GLP-1 were measured by ELIZA in plasma samples. Participants were also divided into three groups based on BMI. Logistic regression models and general linear regression models were used to assess the association between CLOCK genotype and study parameters. Univariate linear regression models were used to assess the interaction between CLOCK and VAS, Food timing, chronotype and sleep on food intakes. After controlling for confounding factors, there was a significant difference between genotypes for physical activity (P = 0.001), waist circumference (P˂0.05), BMI (˂0.01), weight (P = 0.001), GLP-1 (P = 0.02), ghrelin (P = 0.04), appetite (P˂0.001), chronotype (P˂0.001), sleep (P˂0.001), food timing (P˂0.001), energy (P˂0.05), carbohydrate (P˂0.05) and fat intake (P˂0.001). Our findings also show that people with the minor allele C who ate lunch after 3 PM and breakfast after 9 AM are more prone to obesity (P˂0.05). furthermore, there was significant interactions between C allele carrier group and high appetite on fat intake (Pinteraction = 0.041), eat lunch after 3 PM on energy intake (Pinteraction = 0.039) and morning type on fat intake (Pinteraction = 0.021). Sleep reduction, changes in ghrelin and GLP-1 levels, changes in eating behaviors and evening preference that characterized CLOCK 3111C can all contribute to obesity. Furthermore, the data demonstrate a clear relationship between the timing of food intake and obesity. Our results support the hypothesis that the influence of the CLOCK gene may extend to a wide range of variables related to human behaviors.


Denervation causes fiber atrophy and myosin heavy chain co-expression in senescent skeletal muscle.
Although denervation has long been implicated in aging muscle, the degree to which it is causes the fiber atrophy seen in aging muscle is unknown. To address this question, we quantified motoneuron soma counts in the lumbar spinal cord using choline acetyl transferase immunhistochemistry and quantified the size of denervated versus innervated muscle fibers in the gastrocnemius muscle using the in situ expression of the denervation-specific sodium channel, Nav₁.₅, in young adult (YA) and senescent (SEN) rats. To gain insights into the mechanisms driving myofiber atrophy, we also examined the myofiber expression of the two primary ubiquitin ligases necessary for muscle atrophy (MAFbx, MuRF1). MN soma number in lumbar spinal cord declined 27% between YA (638±34 MNs×mm⁻¹) and SEN (469±13 MNs×mm⁻¹). Nav₁.₅ positive fibers (1548±70 μm²) were 35% smaller than Nav₁.₅ negative fibers (2367±78 μm²; P<0.05) in SEN muscle, whereas Nav₁.₅ negative fibers in SEN were only 7% smaller than fibers in YA (2553±33 μm²; P<0.05) where no Nav₁.₅ labeling was seen, suggesting denervation is the primary cause of aging myofiber atrophy. Nav₁.₅ positive fibers had higher levels of MAFbx and MuRF1 (P<0.05), consistent with involvement of the proteasome proteolytic pathway in the atrophy of denervated muscle fibers in aging muscle. In summary, our study provides the first quantitative assessment of the contribution of denervation to myofiber atrophy in aging muscle, suggesting it explains the majority of the atrophy we observed. This striking result suggests a renewed focus should be placed on denervation in seeking understanding of the causes of and treatments for aging muscle atrophy.


Kazal-type chymotrypsin inhibitor from duck pancreas.
A chymotrypsin inhibitor of the Kazal-type has been isolated from duck pancreas, by affinity chromatography on immobilized chymotrypsin, gel filtration on Bio-Gel P-10 and reverse phase (RP)-HPLC. It inhibits bovine chymotrypsin Aalpha with an association constant (K(a)) of 2.06x10(7) M(-1). The complete amino acid sequence was determined after digestion of pyridylethylated inhibitor with Staphylococcus aureus V8 protease and chemical cleavage with CNBr. Duck pancreatic chymotrypsin inhibitor (DPCI) was found to be a single polypeptide chain composed of 65 amino acid residues, corresponding to a molecular mass of 7191 Da.


Herbal medicine Sho-saiko-to (TJ-9) increases expression matrix metalloproteinases (MMPs) with reduced expression of tissue inhibitor of metalloproteinases (TIMPs) in rat stellate cell.
We have reported that Sho-saiko-to (TJ-9) prevents liver fibrosis in vivo. To gain further insights into the effect of TJ-9, the matrix metalloproteinases (MMPs)/tissue inhibitors of metalloproteinases (TIMPs) balance was examined. Hepatic stellate cells (HSCs) were isolated from male Wistar rats and cultured with TJ-9 (0-1000 microg/ml) on uncoated plastic dishes for 4 days. To elucidate the effects on the MMPs/TIMPs balance by TJ-9, quantitative analysis of type IV collagen-degrading activity, gelatin zymography and reverse zymography were carried out. Northern blot analysis was performed to determine the expression of MMP-2, 13 and TIMP-1 mRNAs. TJ-9 treatment resulted in dose-dependent upregulation of MMP-2, 13 mRNA and downregulation of TIMP-1 mRNA up to 500 microg/ml. Gelatin zymography, reverse zymography and quantitative analysis of type IV collagen-degrading activity confirmed that TJ-9 increased MMP-2 activity and prevented TIMP-1, 2 activities in a dose-dependent manner. SB203580 diminished the reduction of mRNA as well as the activity of TIMP-1 by TJ-9 and induction of mRNA as well as the activity of MMP-2. These results show that TJ-9 increased MMP-2, 13 activity with reduced TIMP-1, 2 activities on HSCs possibly via P38 pathway.


Subsequent fracture risk of women with pregnancy and lactation-associated osteoporosis after a median of 6 years of follow-up.
Almost a quarter of patients with PAO will sustain a subsequent fracture; patients need to be informed about potential risks before deciding for further pregnancies. Pregnancy and lactation-associated osteoporosis (PAO) is a severe type of premenopausal osteoporosis which predominantly occurs in the last trimester of pregnancy or immediately postpartum. Long-term follow-up data including subsequent fracture risk have yet to be reported. This single-center prospective cohort study investigated the subsequent fracture risk of all 107 patients with PAO who were referred to our institution. Overall, 107 presented with at least one fracture. Each patient sustained on average four fractures most commonly at the thoracolumbar spine. During a median of 6 years of follow-up, 26 (24.3%) of patients who had a fracture at baseline reported a subsequent fracture. Overall, 30 PAO patients (28%) reported a further pregnancy. In subsequent pregnancies, 6 (20%) of patients reported a subsequent fracture. Patients with up to 1 vs. > 1 fracture at time of diagnosis showed a 3 (10%) and 25 (27%) subsequent fracture rate, respectively (p = 0.047). There was a significant correlation between the number of fractures at time of diagnosis and subsequent fracture risk (N = 26,p= 0.56, p = 0.003). Almost a quarter of patients with PAO will sustain a subsequent fracture, and this fracture risk correlates with the number of fractures at time of diagnosis. Patients with PAO need to be informed about their potential subsequent fracture risk before deciding for further pregnancies.


Effect of a monounsaturated diet vs. a polyunsaturated fatty acid-enriched diet on blood pressure in normotensive women and men.
The effect on blood pressure of monounsaturated and (n-6)polyunsaturated fatty acids was studied under strict dietary control in normotensive subjects. For 17 days 31 women and 27 men received a control diet providing 19.3% of energy as saturated fat. Then subjects were randomized over two test diets: one diet provided 15.1% of energy from monounsaturated and 7.9% from polyunsaturated fatty acids ('mono diet'), and the other diet provided 10.8% from monounsaturated and 12.7% from polyunsaturated fatty acids ('poly diet'). Saturated fat intake was now 12.8% on both diets. Mean blood pressure at the end of the control period was 116/69 mmHg for the mono group and 117/73 mmHg for the poly group. After 5 weeks on the test diet, blood pressure was 115/67 mmHg for the mono group and 117/72 mmHg for the poly group (difference in changes between the two diet groups was not significant). These findings suggest that at a high fat intake, linoleic acid, when providing more than 7.9% of energy intake, does not influence blood pressure relative to oleic acid in normotensive women and men.


Bigenotypic and biphenotypic appearance of immunoglobulin light chains in 1 of 3 non-Hodgkin's lymphoma xenografts.
Chromosomal analysis of three human non-Hodgkin's lymphoma (NHL) xenografts revealed common extra 14q+ segment and other individual clonal markers without involvement of immunoglobulin light-chain regions. Two NHL xenografts (HT 58 and HT 130) showed restricted lambda light chain expression and deleted kappa and rearranged lambda genes. However, in HT 117 both kappa and lambda chains were present on the cell surface, and both light chain genes showed rearranged alleles. This unique phenomenon could have resulted from either disturbed regulation of isotype exclusion or from the malignant transformation of light chain 'uncommitted' B cells.


Probiotic VSL#3 prevents antibiotic-associated diarrhoea in a double-blind, randomized, placebo-controlled clinical trial.
Antibiotic-associated diarrhoea (AAD) is a frequent complication of systemic antibiotic therapy and Clostridium difficile-associated diarrhoea (CDAD) is its most serious form due to associated morbidity and mortality. This trial aimed to investigate whether the probiotic VSL#3 prevents AAD and CDAD in average-risk hospital patients. Adult hospital inpatients exposed to systemic antibiotics were recruited to this multicentre, randomized, double-blind, placebo-controlled trial. One sachet of VSL#3 or placebo was given twice daily for the length of the antibiotics course and for seven days thereafter. Primary outcomes were AAD and CDAD. Patients randomized to active (N = 117) and placebo (N = 112) groups were well-matched for baseline demographic patient data. No cases of CDAD were detected. The rate of AAD was significantly lower in the active group on per protocol analysis (0% active vs 11.4% placebo; P = 0.006). On intention-to-treat analysis the difference in AAD incidence (4.3% active vs 8.9% placebo; P = 0.19) was not significant. VSL#3 is associated with a significant reduction in the incidence of AAD in average-risk hospital inpatients exposed to systemic antibiotics. As the incidence of CDAD has fallen sharply, no cases of CDAD were found. Probiotic administration as prophylaxis for CDAD may not be indicated in average-risk hospital patients.


Amorphous solid dispersions: An update for preparation, characterization, mechanism on bioavailability, stability, regulatory considerations and marketed products.
Amorphous solid dispersions (ASDs) are being employed frequently to improve bioavailability of poorly soluble molecules by enhancing the rate and extant of dissolution in drug product development process. These systems comprise of an amorphous active pharmaceutical ingredient stabilized by a polymer matrix to provide enhanced stability. This review discussed the methodologies of preparation and characterization of ASDs with an emphasis on understanding and predicting stability. Rational selection of polymers, preparation techniques with its advantages and disadvantages and characterization of polymeric amorphous solid dispersions have discussed. Stability aspects have been described as per ICH guidelines which intend to depend on selection of polymers and preparation methods of ASD. The mechanism involved on improvement of bioavailability also considered. Regulatory importance of ASD and current evolving details of QBD approach were reviewed. Amorphous products and particularly ASDs are currently most emerging area in the pharmaceutical field. This strategic approach presents huge impact and advantageous features concerning the overall improvement of drug product performance in clinical settings which ultimately lead to drug product approval by leading regulatory agencies into the market.


Computed tomography analysis of fascial space involvement demonstrates correlations with laboratory tests, length of hospital stays and admission to the intensive care unit in odontogenic infections.
Odontogenic infections are frequent and can spread, leading to complications such as sepsis and the need for admission to an Intensive Care Unit (ICU). The purpose of this study was to perform a computed tomography analysis of the fascial space involvement and correlate with personal data, laboratory tests, length of hospital stays and admission to the ICU in patients with odontogenic infections who required hospitalization. Patients with odontogenic infections admitted between June 2017 and May 2018 were prospectively evaluated. The fascial spaces involved were studied using computed tomography with contrast. The possible correlations of tomographic findings with comorbidities, nutritional status, the causative tooth, laboratory tests, length of hospital stays and admission to the ICU were analyzed. We identified 66 cases of odontogenic infections which were admitted in the period analyzed. The involvement of primary spaces (86.7%) predominated, followed by secondary ones (8.7%), and cervical (4.6%). The most frequently involved fascial spaces were submandibular (27.1%), buccal (20.8%), sublingual (18.3%), submental (16.7%), and others (17.1%). There were significant differences between laboratory tests (p < 0.001), mean hospital stays (p < 0.001), and admissions to the ICU (p < 0.001) depending on the number of fascial spaces involved. There was no influence of comorbidities, nutritional status, or causative tooth on fascial space involvement. There was a relationship between greater involvement of fascial spaces assessed by computed tomography and higher values of laboratory tests, more extended hospitalization stays and admission to the ICU. Level 2b.


Exsanguinating hemorrhage from peripheral varicosities.
A 48-year-old woman presented to the emergency department in cardiovascular collapse after the onset of spontaneous bleeding from a noninflamed, nontraumatized varicose vein of the lower extremity. Despite successful immediate resuscitation she later succumbed to the sequelae of hemorrhagic shock. There was no evidence of coagulopathy or coagulation defect. No cardiovascular disease was found on autopsy. Although not common, spontaneous rupture of peripheral varicosities occurs often enough to be of concern. The pathologic lesions have been classified as acute and chronic, with acute lesions occurring in otherwise normal skin. Exsanguination from this source is not mentioned in standard texts, and only 27 cases have been reported in the literature.


Chemically Programmed Vaccines: Iron Catalysis in Nanoparticles Enhances Combination Immunotherapy and Immunotherapy-Promoted Tumor Ferroptosis.
Immunotherapy has yielded impressive results, but only for a minority of patients with cancer. Therefore, new approaches that potentiate immunotherapy are a pressing medical need. Ferroptosis is a newly described type of programmed cell death driven by iron-dependent phospholipid peroxidation via Fenton chemistry. Here, we developed iron oxide-loaded nanovaccines (IONVs), which, chemically programmed to integrate iron catalysis, drug delivery, and tracking exploiting the characteristics of the tumor microenvironment (TME), improves immunotherapy and activation of ferroptosis. The IONVs trigger danger signals and use molecular disassembly and reversible covalent bonds for targeted antigen delivery and improved immunostimulatory capacity and catalytic iron for targeting tumor cell ferroptosis. IONV- and antibody-mediated TME modulation interfaced with imaging was important toward achieving complete eradication of aggressive and established tumors, eliciting long-lived protective antitumor immunity with no toxicities. This work establishes the feasibility of using nanoparticle iron catalytic activity as a versatile and effective feature for enhancing immunotherapy.


Imaging of Fontan-associated liver disease.
The Fontan operation has dramatically altered the natural history of functionally single ventricle congenital heart disease. Patients who have undergone the Fontan operation are living longer and, thus, noncardiac morbidity resulting from the Fontan operation is increasingly being recognized. Fontan-associated liver disease (FALD), one of the chief morbidities following the Fontan operation, is believed to be a multifactorial process that manifests as hepatic congestion and fibrosis, portal hypertension, and development of focal liver lesions, including malignant tumors. This article reviews the imaging findings of FALD in the pediatric and young adult population, reviews the literature related to the imaging of FALD and discusses possible screening algorithms for this population. The need for further research to better understand the causes of FALD, to establish if early liver stiffness measurements (or their change over time) predict long-term outcomes and complications, and to define optimal liver screening procedures is highlighted.


A rapid method for analysis of the products of in vitro translation of messenger RNA.
A rapid and sensitive method for assaying the production of a specific protein during translation of messenger RNA is described. The method involves running the samples mixed with carrier protein in antibody-containing agarose and determining the radioactivity in the rocket-shaped immunoprecipitates which form. The radioactivity detected is shown to be qualitatively and quantitatively dependent on the amount of immunoreactive protein produced in translation and can thus be used as a measure of specific protein production. The ability of the method to rapidly analyze many samples in parallel is of particular use in screening large numbers of recombinant clones for the ability to bind a specific messenger RNA and affect its translation in vitro.


Pathway-level multi-omics analysis of the molecular mechanisms underlying the toxicity of long-term tacrolimus exposure.
Tacrolimus (TAC)-based treatment is associated with nephrotoxicity and hepatotoxicity; however, the underlying molecular mechanisms responsible for this toxicity have not been fully explored. This study elucidated the molecular processes underlying the toxic effects of TAC using an integrative omics approach. Rats were sacrificed after 4 weeks of daily oral TAC administration at a dose of 5 mg/kg. The liver and kidney underwent genome-wide gene expression profiling and untargeted metabolomics assays. Molecular alterations were identified using individual data profiling modalities and further characterized by pathway-level transcriptomics-metabolomics integration analysis. Metabolic disturbances were mainly related to an imbalance in oxidant-antioxidant status, as well as in lipid and amino acid metabolism in the liver and kidney. Gene expression profiles also indicated profound molecular alterations, including in genes associated with a dysregulated immune response, proinflammatory signals, and programmed cell death in the liver and kidney. Joint-pathway analysis indicated that the toxicity of TAC was associated with DNA synthesis disruption, oxidative stress, and cell membrane permeabilization, as well as lipid and glucose metabolism. In conclusion, our pathway-level integration of transcriptome and metabolome and conventional analyses of individual omics profiles, provided a more comprehensive picture of the molecular changes resulting from TAC toxicity. This study also serves as a valuable resource for subsequent investigations aiming to understand the mechanism underlying the molecular toxicology of TAC.


Child and caregiver psychosocial functioning in pediatric immunodeficiency disorders.
Children with chronic illness have been found to be at an increased risk of behavioral and emotional difficulties. To date, children with pediatric immunodeficiency disorders (PIDDs) and their families have not been the focus of extensive published psychosocial research. To determine if children with PIDDs and their caregivers have altered psychosocial function and whether the severity of the PIDD was associated with such difficulties. Twenty children with PIDDs and 20 children with asthma were recruited for this study. Children and their caregivers completed various psychosocial questionnaire forms. Responses were compared with normative data for the appropriate measure and with other variables. Higher frequencies of children with PIDDs were found to have a number of elevated psychosocial concerns when contrasted with normative data, particularly from parent report. These concerns included depression, anxiety, somatization, social withdrawal, and social skills. The severity of the PIDDs was significantly associated with a number of behavioral adjustment issues, including receiving psychiatric diagnoses and special education services. Although children with PIDDs had significantly more psychiatric diagnoses than did asthmatic children, these groups did not differ significantly on questionnaire scores regarding child or caregiver psychosocial adjustment. Children with PIDDs have significant behavioral problems. Children receiving intravenous immunoglobulin or immunomodulatory treatments were reported to have more problems than children not receiving them. This study highlights the need for further research in psychosocial functioning of children with PIDDs in an effort to develop interventions to promote their overall adjustment.


Effective Radiation Dose Reduction in Computed Tomography With Iterative Reconstruction in Patients With Urinary Stone.
This study aimed to prospectively compare the image quality and visibility of urinary stone on computed tomographic (CT) images at multiple radiation exposure levels from the same patient reconstructed with sinogram-affirmed iterative reconstruction (SAFIRE). This study included 760 patients with urinary stone who underwent CT with simultaneous acquisition at 6 exposures per patient (100% filtered back projection, 75%, 50%, 37.5%, 25%, and 12.5% SAFIRE). Two radiologists independently assessed overall image quality, noise, and stone visibility by using a 5-point scale. Quantitative measurements, including the CT number, image noise, signal-to-noise ratio, contrast-to-noise ratio (CNR), and corresponding figure of merit (FOM), were compared for 100% versus 5 other radiation doses. Qualitative overall image quality, noise, and stone visibility according to the location were not inferior at 37.5% exposure compared with 100% exposure, except for the visualization of smaller stones <3 mm. The signal-to-noise ratio and CNR of CT images were increased at 50% exposure compared with 100% exposure. Computed tomographic images at 37.5% exposure reconstructed with SAFIRE had significantly more noise and a lower CNR compared with CT images reconstructed with filtered back projection, based on FOMnoise and FOMCNR. The size-specific dose estimation was 4.1 ± 0.8 mGy at 37.5% exposure. Computed tomography performed at 37.5% exposure with SAFIRE may be diagnostically acceptable for the detection of clinically relevant stone.


Influence of osmolarity and the presence of an osmoprotectant on lactococcus lactis growth and bacteriocin production
Growth inhibition of Lactococcus lactis provoked by increasing osmolarity is reversed when glycine betaine (GB) or its analogs are added to a defined medium. Lacticin 481 production increased sharply with growth medium osmolarity in the absence of osmoprotectant but remained unaffected when GB was supplied in media of increasing osmolarity.


Development of a mutant of Trichoderma citrinoviride for enhanced production of cellulases.
Considering importance of a microbial strain capable of increased cellulases production and insensitive to catabolite repression for industrial use, we have developed a mutant strain of Trichoderma citrinoviride by multiple exposures to EMS and ethidium bromide. The mutant produced 0.63, 3.12, 8.22 and 1.94 IU ml(-1) FPase, endoglucanase, beta-glucosidase and cellobiase, respectively. These levels were, respectively, 2.14, 2.10, 4.09 and 1.73 fold higher than those in parent strain. Glucose (upto 20 mM) did not repress enzyme production by the mutant under submerged fermentation conditions. In vitro activity assay with partially purified cellulase showed lack of inhibition by glucose. Interestingly, the partially purified endoglucanase and beta-glucosidase were activated by 2.0 fold and 2.6 fold, respectively, by 20 mM and 30 mM ethanol in the assay mixture. Genetic distinction of the mutant was revealed by the presence of two unique amplicans in comparative DNA fingerprinting performed using 20 random primers.


Police-reported intimate partner violence during pregnancy: who is at risk?
The objective of this retrospective cohort study was to determine risk factors for police-reported intimate partner violence (IPV) during pregnancy among Seattle residents with a registered live birth or fetal death in Washington State. Logistic regression was used to calculate adjusted odds ratios (aOR) and 95% confidence intervals for the association between demographic, behavioral, and obstetric history risk factors and any, physical, and non-physical police-reported IPV. Significant risk factors for any police-reported IPV during pregnancy included unmarried status (aOR 2.36), public health program use (aOR 1.33), smoking or alcohol use during pregnancy (aORs 1.45 and 1.80, respectively), previous live birth (aOR 1.39), and previous spontaneous or induced abortion (aORs 1.39 and 1.34, respectively). Risk factors for physical IPV varied only slightly from those for any IPV, and fewer factors were associated with nonphysical IPV. Demographic, behavioral, and obstetric history risk factors are potential markers of IPV risk during pregnancy.


Alterations within pp59v-rel-containing protein complexes following the stimulation of REV-T-transformed lymphoid cells with zinc.
pp59v-rel exists in association with specific cellular proteins within lymphoid cells transformed by reticuloendotheliosis virus (REV-T). These include the cellular rel homolog (p75c-rel) as well as a 40-kDa avian homolog to I kappa B. The brief exposure of REV-T-transformed lymphoid cells to micromolar concentrations of ZnSO4 induces profound alterations within these protein complexes. Most of the constituents of the rel protein complexes (to include pp59v-rel, p75c-rel, and p115) translocate from the cytosol to the nucleus. This system has been used to characterize the molecular events that accompany the activation of rel protein complexes. The level of phosphorylation increases on three proteins within these complexes: pp59v-rel, p75-c-rel, and pp40. The degree of phosphorylation on pp59v-rel is such that its relative mass increases 3 to 6 kDa when resolved by SDS-polyacrylamide gel electrophoresis. pp59v-rel is phosphorylated on serine and threonine residues predominantly within a single domain of 17.5 kDa. Similarly, p75c-rel exhibits a corresponding increase in its relative mass with increased phosphorylation. The increased phosphorylation of pp40 is accompanied by its dissociation from the cytosolic rel protein complexes. These observations draw parallels with the induction of the NF-kappa B trans-activating factor.


A cryptic tubulin-binding domain links MEKK1 to curved tubulin protomers.
The MEKK1 protein is a pivotal kinase activator of responses to cellular stress. Activation of MEKK1 can trigger various responses, including mitogen-activated protein (MAP) kinases, NF-κB signaling, or cell migration. Notably, MEKK1 activity is triggered by microtubule-targeting chemotherapies, among other stressors. Here we show that MEKK1 contains a previously unidentified tumor overexpressed gene (TOG) domain. The MEKK1 TOG domain binds to tubulin heterodimers-a canonical function of TOG domains-but is unusual in that it appears alone rather than as part of a multi-TOG array, and has structural features distinct from previously characterized TOG domains. MEKK1 TOG demonstrates a clear preference for binding curved tubulin heterodimers, which exist in soluble tubulin and at sites of microtubule polymerization and depolymerization. Mutations disrupting tubulin binding decrease microtubule density at the leading edge of polarized cells, suggesting that tubulin binding may play a role in MEKK1 activity at the cellular periphery. We also show that MEKK1 mutations at the tubulin-binding interface of the TOG domain recur in patient-derived tumor sequences, suggesting selective enrichment of tumor cells with disrupted MEKK1-microtubule association. Together, these findings provide a direct link between the MEKK1 protein and tubulin, which is likely to be relevant to cancer cell migration and response to microtubule-modulating therapies.


Pain through the perspective of art and creativity: insights from the Unmasking Pain project.
People struggle to tell their story of living with pain and when they do it is articulated in a way that may not be understood, heard or taken seriously. Unmasking Pain is an artist-led project that explored creative approaches to tell stories of life with pain. The project was led by a dance theatre company that specialises in storytelling and emotional experiences for players and audiences. The project involved artists and people living with ongoing pain co-creating activities and environments to curiously explore "oneself", through imagination and creative expression. This article discusses insights and perspectives emerging from the project. The project revealed the power of art to make-sense of oneself with or without pain, and how art facilitates expression of complex inner experience and personal stories. People described Unmasking Pain as "explorative joy despite pain", and "a new set of rules" that contrasts with those experienced during clinical encounters. We discuss how art has the potential to improve clinical encounters and promote health and well-being, and whether artist-led activities are an intervention, therapy, or something else. Pain rehabilitation specialists from the project described Unmasking Pain as "freeing-up thinking", allowing conceptual thought beyond the biopsychosocial model of pain. We conclude that art has the potential to shift people living with pain from "I can't do, I am not willing to do it" to "Perhaps I can, I'll give it a go, I enjoyed".


Primary synovial sarcoma of the kidney: corroboration with in situ polymerase chain reaction.
Synovial sarcoma is a tumor of the soft tissues with a unique chromosomal translocation t(X;18)(p 11.2;q11.2) that can be detected by polymerase chain reaction in tissue homogenates. The case of a 32-year-old woman with a primary synovial sarcoma of the kidney is described, the diagnosis was corroborated by the recently developed method of in situ polymerase chain reaction (IS-PCR). Synovial sarcoma of the kidney may be confused with other spindle cell tumors, for that reason IS-PCR may be useful to confirm the diagnosis in paraffin-embedded material.


The role of CARD15 mutations and smoking in the course of Crohn's disease in a Mediterranean area.
To evaluate the role of CARD15 mutations and smoking in the main events of Crohn's disease (CD). A total of 182 patients with CD were included in a prospective study in order to evaluate the role of CARD15 mutations and smoking in the main outcomes of disease course: first operation and surgical recurrence. The following variables were evaluated in a univariable and multivariable analysis: age, sex, site of disease, pattern, smoking habit, extraintestinal manifestations, duration of disease, and CARD15 mutation. The Kaplan-Meier method for survival curves and Cox model for multivariable analysis were, respectively, used. A total of 110 patients were operated on and 32 were reoperated on. The 7-yr cumulative free rate of surgery was 42% (95% CI 34-51%). At multivariate analysis only stricturing and penetrating pattern were predictors of surgery (HR 1.7, 95% CI 1-2.8; HR 3.2, CI 1.8-5.5, respectively). The 7-yr cumulative free rate of reoperation was 75% (95% CI 0.52-0.88). At multivariable analysis in the model with any CARD15 mutation, only smoking habit at diagnosis (HR 3.6, 95% CI 1.4-9.1) was predictive of surgical recurrence. When single mutations were considered in the model smoking (HR 4.2, 95% CI 1.8-10.1) and L1007fs mutation (HR 2.9, 95% CI 1.1-7.3) were predictive of reoperation. In CD, smoking predicts recurrence after surgery. The role of CARD15 mutations in the clinical course of CD remains undefined.


Intrahepatic extramedullary hematopoietic tumor mimicking metastatic carcinoma from a colonic primary.
Extramedullary hematopoiesis (EMH) is associated with a number of diseases in which the normal function of the bone marrow is disturbed. While organs with hemopoietic capacity like the liver and spleen are most commonly involved, EMH has also occasionally been found in other organs like the adrenal gland, lymph nodes, breast, thymus, small bowel and central nervous system. However, presentation of a myeloproliferative disorder, such as EMH in these organs is a rare event. We report clinical and fine-needle aspiration (FNA) findings in a patient who presented with intrahepatic EMH which mimicked metastatic carcinoma from a colonic primary. Ultrasound-guided FNA of the intrahepatic mass revealed megakaryocytes and myelocytes thus establishing the diagnosis of EMH. EMH is an unusual condition that can mimic other solid masses of the liver. Because radiologic findings are not specific, EMH should be considered in the differential diagnosis, especially in patients with a myeloproliferative disorder. FNA and subsequent cytopathological interpretation of the aspirates enables avoidance of unnecessary potentially hazardous surgery.


The effects of knee immobilization on marrow adipocyte hyperplasia and hypertrophy at the proximal rat tibia epiphysis.
Marrow adipose deposition is observed during aging and in association with extended periods of immobility. The objective of this study was to determine the contribution of adipocyte hypertrophy and hyperplasia to bone marrow fat deposition induced by immobilization of the rat knee joint for 2, 4, 16 or 32 weeks. Histomorphometric analyses compared immobilized to sham-operated proximal tibia from age and gender matched rats to assess the contribution of aging and duration of immobilization on the number and size of marrow adipocytes. Results indicated that marrow adipose tissue increased with the duration of immobilization and was significant larger at 16 weeks compared to the sham-operated group (0.09956±0.13276mm2 vs 0.01990±0.01100mm2, p=0.047). The marrow adipose tissue was characterized by hyperplasia of adipocytes with a smaller average size after 2 and 4 weeks of immobilization (at 2 weeks hyperplasia: 68.86±33.62 vs 43.57±24.47 adipocytes/mm2, p=0.048; at 4 weeks hypotrophy: 0.00036±0.00019 vs 0.00046±0.00023mm2, p=0.027), and by adipocyte hypertrophy after 16 weeks of immobilization (0.00083±0.00049 vs 0.00046±0.00028mm2, p=0.027) compared to sham-operated. Both immobilized and sham-operated groups showed marrow adipose conversion with age; immobilized (p=0.008; sham: p=0.003). Overall, fat deposition in the bone marrow of the proximal rat tibia epiphysis and induced by knee joint immobilization was characterized by hyperplasia of small adipocytes in the early phase and by adipocyte hypertrophy in the later phase. Mediators of marrow fat deposition after immobilization and preventive countermeasures need to be investigated.


Induction of Fetal Hemoglobin by Introducing Natural Hereditary Persistence of Fetal Hemoglobin Mutations in the γ-Globin Gene Promoters for Genome Editing Therapies for β-Thalassemia.
Reactivation of γ-globin expression is a promising therapeutic approach for β-hemoglobinopathies. Here, we propose a novel Cas9/AAV6-mediated genome editing strategy for the treatment of β-thalassemia: Natural HPFH mutations -113A > G, -114C > T, -117G>A, -175T > C, -195C > G, and -198T > C were introduced by homologous recombination following disruption of BCL11A binding sites in HBG1/HBG2 promoters. Precise on-target editing and significantly increased γ-globin expression during erythroid differentiation were observed in both HUDEP-2 cells and primary HSPCs from β-thalassemia major patients. Moreover, edited HSPCs maintained the capacity for long-term hematopoietic reconstitution in B-NDG hTHPO mice. This study provides evidence of the effectiveness of introducing naturally occurring HPFH mutations as a genetic therapy for β-thalassemia.


The effect of anterior ankle-foot orthosis and posterior ankle-foot orthosis on functional ambulation in stroke patients.
The goal of rehabilitation after stroke is to restore safe and sufficient function to hemiplegic patients, and prescription of an ankle-foot orthosis (AFO) to improve speed and functional ambulation is a part of this program. This crossover randomized interventional study aimed to evaluate the effect of an anterior ankle-foot orthosis (AAFO) and posterior leaf-spring ankle-foot orthosis (PLS AFO) on speed and functional ambulation in hemiplegic stroke patients. Clinical assessments were performed on 11 hemiplegic stroke patients by the AAFO, PLS AFO, and wearing shoes. Functional ambulation was measured by the 6-min walking test, Timed Up and Go Test, Time Up and Down Stair Test, and Functional Ambulation Category. Walking speed was measured by the 10-m test. Both PLS AFO and AAFO significantly improved the performance of TUDS and TUG tests in hemiplegic patients. However, by using PLS AFO, walking distance was significantly greater than walking with shoes. There was no significant effect on the walking speed improvement using PLS AFO or AAFO compared to wearing shoes. The positive effects of the AAFO and PLS AFO on functional ambulation were significant. By using PLS AFO, hemiplegic patients could walk a longer distance than wearing shoes.


Measuring the environment for friendliness toward physical activity: a comparison of the reliability of 3 questionnaires.
We tested the reliability of 3 instruments that assessed social and physical environments. We conducted a test-retest study among US adults (n = 289). We used telephone survey methods to measure suitableness of the perceived (vs objective) environment for recreational physical activity and nonmotorized transportation. Most questions in our surveys that attempted to measure specific characteristics of the built environment showed moderate to high reliability. Questions about the social environment showed lower reliability than those that assessed the physical environment. Certain blocks of questions appeared to be selectively more reliable for urban or rural respondents. Despite differences in content and in response formats, all 3 surveys showed evidence of reliability, and most items are now ready for use in research and in public health surveillance.


Perioperative Medicine: The Value Proposition for Anesthesia?: A UK Perspective on Delivering Value from Anesthesiology.
Perioperative medicine describes the practice of patient centered, multidisciplinary, and integrated medical care of patients from the moment of contemplation of surgery until full recovery. The value proposition for perioperative medicine rests on defining benefits that outweigh the costs of change. This article discusses the concept of value in the context of healthcare and highlights a number of reasons for relative market failure. Five key opportunities for adding value in the perioperative journey are suggested: collaborative decision-making, lifestyle modification before surgery, standardization of in-hospital perioperative care, achieving full recovery after surgery, and the use of data for quality improvement.


A simple cryogenic holder for tensile testing of soft biological tissues.
To overcome the difficulty of gripping soft biological materials for tensile test, a simple inexpensive cryogenic holder was developed which allows rapid (3 min) preparation of samples. It is made of 6 parts, built in a bakelite cloth, which is an excellent thermal isolant, and is used with rectangular (8x10(-2)x10(-2)x10(-2)m) samples. The holder with the sample inside is completely immersed in liquid nitrogen for 50 s. This duration allows the freezing of the sample ends on a 10(-2)m length and gives a very flat freezing surface throughout the sample cross section. The 6x10(-2)m central part of the sample remained at ambient temperature. Two parts of the holder help maintain the sample until its ends are vertically gripped in the tensile machine thus avoiding any sample deformation during this step. No pressure was applied on the frozen part of the sample by grips of the tensile machine and this avoids breaks in this region. The sample is fixed by adhesion forces (>1 kN) between its frozen parts and 2 pieces of the holder. The procedure has been successfully tested with bovine and salmon muscle samples and results show tensile breaks randomly distributed in the unfrozen region of the samples. Particular attention has been paid to obtain a very flat freezing surface so that the axial strain is equal throughout the sample and therefore any strain-related mechanical parameters can be accurately determined. The dimensions of the holder can be easily modified to fit other sample geometries and can be used with other biological materials.


Bacteriophage-borne enzymes in carbohydrate chemistry. Part I. On the glycanase activity associated with particles of Klebsiella bacteriophage No. 11.
The preparation and use of particles of Klebsiella bacteriophage No. 11 are described. A glycanase activity associated with the viruses catalyses the depolymerization of (alkali-treated) Klebsiella serotype 11 capsular polysaccharide, ultimately to a mixture of oligosaccharides consisting of one or two repeating units. Mainly glucosidic bonds are hydrolysed. The substrate specificity of the viral enzyme has been characterized by using derivatives of serotype-11 polysaccharide, as well as 81 heterologous, bacterial, capsular glycans. It is concluded that the glycanase will (at least) also depolymerize all polysaccharides containing the unsubstituted chain-trisaccharide repeating-unit of its natural substrate.


Exploring the 'Patient Experience' of Individuals with Limited English Proficiency: A Scoping Review.
Individuals with limited English proficiency (LEP) face barriers to safe and high-quality health care. 'Patient-experience' is increasingly viewed as an important component of health care quality. However, the impact of language proficiency on 'patient-experience' is not well-described. This scoping review mapped the literature on the patient experience of individuals with LEP. We reviewed sixty qualitative and mixed-methods studies from EMBASE and MEDLINE published between 2007 and 2017. We identified four major themes: (1) Communication, language barriers, and health literacy, (2) Relationships with health care professionals, (3) Discrimination and intersection with other dimensions of identity, and (4) Cultural safety. We also identified factors that may improve LEP patient experience, including: mitigating language barriers through interpretation or language-concordant providers, offering translated patient resources, and educating health care professionals about cultural safety.


Effects of repeated administration of (-)-nicotine on AF64A-induced learning and memory impairment in rats.
It has been reported that pretreatment with (-)-nicotine prevents glutamate- and amyloid beta protein (Abeta)-induced cytotoxicity in vitro. However, few studies on the neuroprotective effects of (-)-nicotine in vivo have been reported. We examined whether repeated administration of (-)-nicotine exhibits neuroprotective effects in AF64A-treated rats. (-)-Nicotine (0.1 and 0.2 mg/kg, s.c.) was administered once a day for 28 days. On day 14, AF64A (2.5 nmol/side) was injected bilaterally into the hippocampus. Intrahippocampal injection of AF64A showed severe impairment of learning and memory in rats in the water maze and passive avoidance tests. Repeated administration of (-)-nicotine (0.1 and 0.2 mg/kg, s.c.) did not reverse the impairment of memory induced by AF64A in the water maze test. Interestingly, the (-)-nicotine (0.1 and 0.2 mg/kg, s.c.)-treated group showed weak impairment of learning and memory after AF64A treatment compared to the (AF64A + saline)-treated group in the passive avoidance test. These results suggested that (-)-nicotine may have neuroprotective effects against the neurotoxicity induced by AF64A.


[The role of miR-223 in the acute rejection after kidney transplantation].
To explore the role of miR-223 in the acute rejection after kidney transplantation. 33 patients who received kidney transplantation in our hospital within a year were collected and 12 of them appeared acute rejection within 1 month after surgery. In all the patients, the peripheral blood miR-223 level was collected and detected by blind arrangements. Furthermore, PBMCs from healthy volunteers were also collected and stimulated by PHA and then miR-223 level was measured. In the peripheral blood cell, the miR-223 was increased 2 times after acute rejection, as well as 3. 76 times after PHA treatment.Furthermore, using miR-223 predicts AR had a specificity of 90% and sensitivity of 92%. The miR-223 may have significant role in the acute rejection of kidney transplantation.


X-chromosomal monosomy (77,X0) in a Doberman Pinscher with gonadal dysgenesis.
A 6-month-old stunted female Doberman Pinscher was found to have a 77,X0 chromosomal complement. The ovaries were small, consisting primarily of interstitial-type cells and solid epithelial cords. The dam, sire, and a male littermate had normal karyotypes.


Selection and characterization of hepatitis C virus replicons dually resistant to the polymerase and protease inhibitors HCV-796 and boceprevir (SCH 503034).
HCV-796 is a nonnucleoside inhibitor of the hepatitis C virus (HCV) nonstructural protein 5B (NS5B) polymerase, and boceprevir is an inhibitor of the NS3 serine protease. The emergence of replicon variants resistant to the combination of HCV-796 and boceprevir was evaluated. Combining the inhibitors greatly reduced the frequency with which resistant colonies arose; however, some resistant replicon cells could be isolated by the use of low inhibitor concentrations. These replicons were approximately 1,000-fold less susceptible to HCV-796 and 9-fold less susceptible to boceprevir. They also exhibited resistance to anthranilate nonnucleoside inhibitors of NS5B but were fully sensitive to inhibitors of different mechanisms: a pyranoindole, Hsp90 inhibitors, an NS5B nucleoside inhibitor, and pegylated interferon (Peg-IFN). The replicon was cleared from the combination-resistant cells by extended treatment with Peg-IFN. Mutations known to confer resistance to HCV-796 (NS5B C316Y) and boceprevir (NS3 V170A) were present in the combination-resistant replicons. These changes could be selected together and coexist in the same genome. The replicon bearing both changes exhibited reduced sensitivity to inhibition by HCV-796 and boceprevir but had a reduced replicative capacity.


Selective laser trabeculoplasty vs micropulse laser trabeculoplasty in open-angle glaucoma.
The aim of this study was to prospectively compare the efficacy, safety, and tolerability of selective laser trabeculoplasty (SLT) vs micropulse laser trabeculoplasty (MLT) in reducing intraocular pressure (IOP) in open-angle glaucoma patients. In all, 38 patients were randomized to 360° MLT and 31 patients were randomized to 360° SLT. IOP was measured at intervals of 1 hour and 1, 1-6, 6-12, 6-24, 24-36, and 36-52 weeks. Patients completed a survey 1 week after the procedure. Patients with end-stage, neovascular, uveitic, or angle-closure glaucoma were excluded. Treatment response was defined as an IOP reduction of ≥20.0% or ≥3 mmHg from baseline. IOP was lowered to ≥3 mmHg from baseline among 37.0% of the micropulse patients and 36.0% of patients in the selective laser group at 24-52 weeks. Similarly, 29.6% of the micropulse patients and 36.0% of the selective laser patients experienced a 20.0% IOP decrease from baseline during the 24-52-week interval (P=0.77). Both groups revealed similar reductions in IOP as absolute values and percentage decreases from baseline at all intervals up to 52 weeks post treatment. There were more treatment failures in the micropulse group up to 52 weeks post laser treatment; however, this was not statistically significant. The micropulse group reported less pain both during and after the procedure (P=0.005). Micropulse trabeculoplasty has demonstrated similar efficacy to SLT over a 52-week follow-up period with less discomfort experienced both during and after the procedure.