Calafiore cardioplegia offers better myocardial protection in adult and senescent rat hearts than Del Nido cardioplegia.
We compared the cardioprotective capacity of Del Nido cardioplegia and warm Calafiore blood cardioplegia in an experimental setting during 90 min of ischaemia. 20 adult and 20 senescent rat hearts were isolated and mounted on a blood-perfused, pressure-controlled Langendorff apparatus. After a stabilization period, cardiac arrest (90 min) was induced by the administration of either Calafiore (Cala) or Del Nido solution (DNS). While Cala was given warm and intermittently, DNS was given as a cold single shot. During 90 min of reperfusion, cardiac function and metabolism were evaluated and biomarker levels were measured. After the end of the experiment, hearts were prepared for electronmicroscopic investigation. Hearts exposed to Cala recovered faster during reperfusion compared with hearts administered DNS (Cala vs DNS at 30 min reperfusion: left ventricular developed pressure 72, SD: 22% of baseline (BL) versus 40, SD: 32% of BL, p < .001, and positive derived left ventricular pressure over time was better in both adult and senescent Cala groups (96, SD: 31% of BL) than in the DNS groups (39, SD: 27% of BL, p < .001). Ischaemic contractures were seen in the DNS groups starting after 30 min of ischaemia, whereas no rise in diastolic pressure was observed for the Cala groups. Accordingly, lactate production was higher after DNS (1.23 mg/dl (SD 0.87) than after Cala (0.33 mg/dl (SD 0.68), p = .015) at the beginning of reperfusion. Troponin I levels at the end of reperfusion were higher after DNS treatment in adult hearts (DNS: 287.9 SD: 147.7 ng/mL vs Cala 91.2: SD: 94.7 ng/mL, p = .02) and in senescent hearts (DNS: 376.5 (SD: 162.8) ng/ml versus Cala 104.7 (SD: 150.2) ng/ml, p = .025). Electron microscopy showed that the cellular oedema index was higher in adult DNS hearts (1.2 ± 0.2) than in adult Cala hearts (0.8 ± 0.1, p = .012), whereas the VS ratio was similar (0.18 ± 0.01 vs 0.17 ± 0.03). Calafiore cardioplegia offers better myocardial protection from ischaemia/reperfusion-related damage in isolated perfused adult and senescent rat hearts than Del Nido cardioplegia.


Incidence of Functional Nasal Voice in Patients With Patulous Eustachian Tube.
Some patients with a patulous Eustachian tube (PET) complain of a nasal voice. This feature is often dismissed without further investigation. As such, there are only a few reports on this important symptom and scant studies have been conducted on a sufficiently large number of cases with PET. Therefore, this study was undertaken to investigate the characteristics of patients having a nasal voice and to examine whether this symptom can be an indication of the severity of PET. Retrospective. Tertiary referral center. A retrospective survey of medical records in Sen-En Rifu Hospital identified 85 patients (40 men and 45 women) with PET between 2013 and 2016. Diagnosis of definite PET was based on the Proposal on Diagnostic Criteria of PET announced by the Otological Society of Japan (2017). The questionnaire inquired about the presence of a nasal voice and it was distributed to each patient at the first visit to the clinic. If a patient marked "yes" for the presence of nasal voice, he/she was later asked on the telephone to exclude nasal voice ascribable to causes other than PET, such as nasal diseases. Correlation between nasal voice and patient characteristics (age, sex, affected side, and PET symptoms such as autophony of own voice, aural fullness, and autophony of breathing sounds), subjective severity of PET evaluated by patulous Eustachian tube handicap inventory-10 (PHI-10), and that with the objective severity of PET evaluated by tubo-tympano-aerodynamic-graphy (TTAG) and sonotubometry were investigated. Seventy-six patients (36 men and 40 women) with definite PET were evaluated in this study. Thirteen patients (17.1%) (five men and eight women) reported a nasal voice coinciding with the occurrence of PET symptoms such as voice autophony, aural fullness, and breathing autophony. Age, sex, affected side, PET symptoms (autophony of their own voice, aural fullness, and autophony of their breathing sounds), and objective findings (TTAG and sonotubometry) were not significantly different between the two groups. The average total score of the PHI-10 in the "PET associated Nasal Voice Group" was 35.8 ± 4.5, which was statistically higher than that of the "non PET associated Nasal Voice Group" 23.6 ± 10.7 (p = 0.002). Out of 76 patients, 44 were treated surgically (Kobayashi Plug). In the "PET associated Nasal Voice Group," 85% (11 out of 13) were subjected to surgical treatment, whereas 52% (33 out of 63) underwent surgical treatment in the "non PET associated Nasal Voice Group." The rate of surgical treatment was significantly higher in "PET associated Nasal Voice Group" (p = 0.047). Nasal voice due to PET symptoms was observed in 17.1% of PET patients. It was generally found in patients with severe subjective symptoms. Nasal voice can be an indication of subjective severity. However, this study failed to show objective evidence of wider Eustachian tube in such cases. Patients with a nasal voice tended to seek vigorous treatment including surgery.


Consistent group selection in high-dimensional linear regression.
In regression problems where covariates can be naturally grouped, the group Lasso is an attractive method for variable selection since it respects the grouping structure in the data. We study the selection and estimation properties of the group Lasso in high-dimensional settings when the number of groups exceeds the sample size. We provide sufficient conditions under which the group Lasso selects a model whose dimension is comparable with the underlying model with high probability and is estimation consistent. However, the group Lasso is, in general, not selection consistent and also tends to select groups that are not important in the model. To improve the selection results, we propose an adaptive group Lasso method which is a generalization of the adaptive Lasso and requires an initial estimator. We show that the adaptive group Lasso is consistent in group selection under certain conditions if the group Lasso is used as the initial estimator.


Liver albumin synthesis in sepsis in the rat: influence of parenteral nutrition, glutamine and growth hormone.
The effect of sepsis on liver synthesis of albumin remains controversial, with studies in man suggesting that synthesis increases, whereas in animals increased, decreased and unaltered synthesis have been reported. To reconcile these conflicting data, total and relative albumin synthesis was measured in rats 24 h after caecal ligation and puncture (CLP) by immunoprecipitation of albumin following a flooding dose of L-[4-3H]phenylalanine. Following CLP, animals were starved for 18 h and then received intravenous infusions of saline or parenteral nutrition (PN) with or without glutamine for 6 h. In animals receiving PN, parenteral injections of growth hormone (GH) or saline vehicle were also administered. Fractional rate of liver total protein synthesis was elevated and total albumin synthesis rate was reduced in all CLP groups when compared with non-operated animals. Total albumin synthesis was also lower in all animals receiving PN than those receiving saline alone, although these differences did not attain statistical significance, except for the group receiving PN+GH. Relative albumin synthesis was also reduced after CLP, and was significantly lower in animals receiving PN than in those receiving saline alone. These findings suggest that in sepsis hepatic protein synthesis is reprioritized away from the production of albumin towards the production of acute-phase proteins and that this change is not influenced by the provision of nutritional support, glutamine or the administration of GH.


The organizational and performance effects of nurse practitioner roles.
Most studies evaluating the roles of Nurse Practitioners have compared the care delivered by individual Nurse Practitioners with that provided by other professionals. These studies should be complemented by research focusing on a higher unit of analysis, namely the organization of the care process for a specific patient group. The most important reason is that Nurse Practitioners are increasingly involved in direct, multiprofessional care in complex health care organizations and networks. In these work settings, their roles may, in both positive and negative ways, lead to changes in the organization of the entire care delivery system. The aim of this paper is to stimulate awareness and evaluation of these organizational changes and their potential impact on the effectiveness of the care process. A conceptual model based on patterned systems contingency theory is proposed. With the help of this model, attention is drawn to issues at the level of the organization and the effectiveness of the care processes that merit attention when Nurse Practitioner positions are being introduced. These issues are derived from case studies in Dutch hospitals. According to the model, a Nurse Practitioner position will change the work structure of the care process involved. Therefore, the effectiveness of a Nurse Practitioner position will be dependent on the changes realized in the work structure. The resulting structure should fit the task characteristics of the care services demanded by the specific patient group. On the basis of this model and the examples presented, questions for further study are formulated. Nurse Practitioner roles can only enhance the effectiveness of care processes when embedded in a work structure that is internally consistent and adjusted to the task environment and available skill-mix. A structural contingency framework may be helpful in identifying relevant organizational issues. To determine the effects of Nurse Practitioner roles, cross-sectional as well as longitudinal studies are needed.


Is RIRS emerging as the preferred option for the management of 2 cm-4 cm renal stones: our experience.
The European Association of Urology (EAU) and the American Urological Association (AUA) guidelines recommend percutaneous nephrolithotomy (PCNL) as the first-line treatment of renal stones greater than 20 mm, however multistage retrograde intrarenal stone surgery (RIRS) is reported to have high stone-free rates (SFR), fewer complications and a rapid learning curve. This study presents our experience of RIRS in the management of 2 cm-4 cm renal stones. A retrospective study was performed of all patients who underwent RIRS for 2 cm-4 cm renal stones over a period of 22 months. The demographics of 71 patients as well as the stone and procedural demographics were recorded. Pre and postoperative radiological assessment was performed by NCCT scanning in 83% of the patients and ureteral access sheaths were used in only 12% of the patients. The severity of surgical complications was determined according to the Clavien-Dindo system. RIRS was performed on 71 patients for renal stones with a mean size of 26 mm. The mean number of procedures per patient was 2.1 and the overall SFR was 81%. Few complications were encountered and only 1 patient had III-b Clavien complication. The study further supports RIRS as a safe and efficacious treatment option for renal stones of 2 cm-4 cm in size. Although both the EAU and AUA do not currently recommend RIRS as the first-line treatment of such stones, it appears to be emerging as a commonly utilized primary modality.


Different mechanisms of beta-adrenoceptor down-regulation by chronic imipramine and electroconvulsive treatment: possible role for protein kinase C.
The aim of this study was to find out how protein kinase C (PKC) is involved in down-regulation of the beta-adrenoceptor in cortical slices of rats subjected to antidepressant treatments. The responses of the cyclic AMP generating system to forskolin, isoproterenol, and noradrenaline were tested in the absence and presence of a PKC activator, 12-O-tetradecanoylphorbol 13-acetate (TPA). The antidepressive treatments applied were chronic administration of imipramine and electroconvulsive shock. The potentiating effect of the phorbol ester on cyclic AMP response to isoproterenol was retained in imipramine-treated animals and even accentuated in rats subjected to electroconvulsive treatment; the TPA effect on noradrenaline-induced cyclic AMP response was blunted in rats receiving imipramine, but augmented in those receiving electroconvulsive treatment. In imipramine-treated rats the beta-down-regulation was still evident in the presence of TPA; after electroconvulsive treatment the phorbol ester-induced potentiation was so high that no significant beta-down-regulation could be observed. No procedure affected the response to forskolin. The beta-down-regulation that develops during chronic imipramine treatment differs from that caused by chronic electroconvulsive treatment; in both cases it is not related to the direct effect on adenylate cyclase.


Laparoscopic wedge resection for gastrointestinal stromal tumors of the stomach: initial experience.
Surgery for gastrointestinal stromal tumors (GIST) of the stomach is now frequently performed using a laparoscopic approach. We investigated the feasibility and effectiveness of laparoscopy in the management of GIST of the stomach. We reviewed the records of 12 consecutive patients who underwent laparoscopic surgery for GIST between April 2000 and April 2004, and compared their short-term outcomes with those of patients who underwent open surgery. All laparoscopic wedge resections were done using stapling devices and 3-4 trocars, often with the aid of intraoperative gastroscopy. We examined all patients preoperatively using various diagnostic modalities, including endoscopic ultrasonography-guided fine-needle aspiration (EUS-FNA). A laparoscopic approach was not indicated if the tumor was located near the cardia or pylorus or if it was >=5 cm in diameter. A specific diagnosis of GIST was obtained preoperatively by EUS-FNA in 10 of the 12 patients. The median diameter of the lesion was 2.7 cm (range, 1.5-4.8 cm). Although intraoperative complications were encountered in two patients, conversion to open surgery was not required, and we were able to perform complete tumor excision with negative surgical margins in all patients. The median operative time was 100 min (range, 65-180 min), similar to that for open surgery. First flatus was passed earlier, and the interval to resuming oral intake was shorter than after open surgery. No major postoperative complications such as leakage developed, and the median postoperative hospital stay was 7 days (range, 5-12 days). All diagnoses made by EUS-FNA were confirmed by immunohisto-pathological evaluation of resected specimens. Laparoscopic wedge resection is a feasible treatment option for GISTs of the stomach if the lesion is <5 cm in diameter.


Sensitivity to changes in dynamic affordances for walking on land and at sea.
We investigated the perception of affordances for walking along a narrow path. We asked whether participants could perceive changes in affordances brought about by manipulation of properties of the body, or of the environment, without direct practice of the to-be-perceived affordance, and without external feedback about the accuracy of perception. In Experiment 1, participants made a series of 8 judgments of how far they could walk along a narrow path either, 1) without added weight, 2) while wearing a weighted vest, or 3) while wearing weights on their ankles. Before walking, mean judgments were lower when wearing weights than in the no-weight condition. In addition, in both weight conditions judgments changed across the series of 8 judgments, in the direction of greater accuracy. Control of the body in walking also can be influenced by motion of the ground surface, as commonly happens in vehicles. In Experiment 2, on a ship at sea, we evaluated the effects of walking with or without weight added to the body at the ankles. We again asked participants (experienced maritime crewmembers) to judge how far they could walk along a narrow path, with versus without ankle weights. As in Experiment 1, judgments made before walking mirrored the observed differences in walking performance. In addition, we again found evidence that judgment improved (without walking practice, or feedback) over the series of judgments. We conclude that participants were sensitive to (and spontaneously learned about) how affordances for walking were influenced by changes in the dynamics of body and the environment.


Empirical antifungal therapy in febrile neutropenic patients: current status.
Empirical antifungal therapy has become established as standard practice in hematology and oncology units over the past decade and its use is increasing. A number of agents have been evaluated and intravenous amphotericin B has emerged as the drug of choice. Evidence of its benefit is limited and only clearly demonstrated in patients not receiving prior antifungal prophylaxis. However, although there have been improvements in the diagnosis of invasive fungal infections, it has been well shown that many patients who die during periods of neutropenia succumb to undiagnosed fungal infection, and also, if treatment is to be effective, it should be started as soon as possible after the onset of infection. Better targeting of antifungal prophylaxis (or preemptive therapy) and empirical therapy may now be possible and standard empirical therapy needs to be reevaluated in the light of changes in the underlying immune status of neutropenic patients and the development of new antifungal agents for prophylaxis and treatment.


Comparative responses of bronchial rings to mediators of airway hyperreactivity in healthy horses and those affected with summer pasture-associated obstructive pulmonary disease.
To compare responses of bronchial rings obtained from healthy horses and horses affected with summer pasture-associated obstructive pulmonary disease (SPAOPD) to selected mediators of airway hyperreactivity in vitro. Bronchial rings from 6 healthy horses and 6 horses affected with SPAOPD. Bronchial rings obtained from each group of horses were mounted in organ baths and attached to force transducers interfaced with a polygraph. After applying 2g of tension, each ring was allowed to equilibrate for 45 minutes in Tyrode's solution at 37 C. Cumulative concentration-response relationships to graded concentrations of selected mediators (10(-8) to 10(-4) M) were determined and analyzed for significance at each concentration. Acetylcholine, histamine, 5-hydroxytryptamine, and leukotriene D4 induced concentration-dependent contractile responses in bronchial rings. Prostaglandin F2alpha induced weak and inconsistent contractile responses. The other 2 agents, norepinephrine and substance P, did not induce concentration-dependent responses. Considering the overall group-drug effect, acetylcholine, histamine, 5-hydroxytryptamine, and leukotriene D4 were effective in inducing consistent concentration-dependent contractile responses in both groups. Only 5-hydroxytryptamine and histamine induced significant responses in contractility between groups. The response of bronchial rings from horses with SPAOPD to 5-hydroxytryptamine was significantly greater than those from control horses, whereas the response to histamine was significantly lower. Significant responses were evident at concentrations ranging from 10(-6) to 10(-4) M for both drugs. Because the airways of horses with SPAOPD had increased responsiveness to 5-hydroxytryptamine in vitro, treatment modalities using 5-hydroxytryptamine antagonists should be investigated to address this phenomenon.


Clinicopathologic growth factors in vestibular schwannomas: a morphological and immunohistochemical study of 69 tumours.
Tumour growth of vestibular schwannomas is still difficult to predict. The aim of our study was to determine whether any defined histopathological feature was correlated with the clinical course. We did a retrospective study with 69 paraffin-embedded tumours to establish whether the number of vessels, blood cells extravasation or degree of inflammation, all semi-quantitatively assessed, could be indicative of potential of growth. An immunohistochemical study was also performed with an endothelial marker CD34, the leukocyte common antigen CD45 and the estrogen and progesterone hormone receptors. All these parameters were correlated with patient's age, duration of symptoms (d), with a clinical growth index (CLI = tumour size/d). No clinical parameters proved to be predictive of tumour growth. Tumour size was significantly (p = 0.01) related to the number of vessels and we found a significant relationship between the clinical growth index (CLI) and total number of vessels, especially when duration of symptoms lasted less than 1 year (p < 0.001). However, we found no relationship between duration of symptoms or CLI and CD34 index. The degree of inflammation was significantly correlated (p = 0.007) with duration of symptoms when it lasted more than 1 year. The CD45 index and the semi-quantitative evaluation of the inflammation were well correlated (p = 0.001). No estrogen receptors antigenic site was detected and only seven tumours expressed progesterone receptor in a few cells without any significant clinical value. These results suggest that vessel density is determinant for sporadic acoustic neuroma growth especially for a short clinical course.


HTLV-2 Encoded Antisense Protein APH-2 Suppresses HIV-1 Replication.
Antisense protein of Human T-cell Leukemia Virus Type 2 (HTLV-2), also called APH-2, negatively regulates the HTLV-2 and helps the virus to maintain latency via scheming the transcription. Despite the remarkable occurrence of HTLV-2/HIV-1 co-infection, the role of APH-2 influencing HIV-1 replication kinetics is poorly understood and needs investigation. In this study, we investigated the plausible role of APH-2 regulating HIV-1 replication. Herein, we report that the overexpression of APH-2 not only hampered the release of HIV-1 pNL4.3 from 293T cells in a dose-dependent manner but also affected the cellular gag expression. A similar and consistent effect of APH-2 overexpression was also observed in case of HIV-1 gag expression vector HXB2 pGag-EGFP. APH-2 overexpression also inhibited the ability of HIV-1 Tat to transactivate the HIV-1 LTR-driven expression of luciferase. Furthermore, the introduction of mutations in the IXXLL motif at the N-terminal domain of APH-2 reverted the inhibitory effect on HIV-1 Tat-mediated transcription, suggesting the possible role of this motif towards the downregulation of Tat-mediated transactivation. Overall, these findings indicate that the HTLV-2 APH-2 may affect the HIV-1 replication at multiple levels by (a) inhibiting the Tat-mediated transactivation and (b) hampering the virus release by affecting the cellular gag expression.


Patients' preoperative expectations predict the outcome of rotator cuff repair.
We are not aware of any previous studies of the relationship between patients' expectations regarding rotator cuff repair and the actual outcome. We hypothesized that preoperative expectations are predictive of the outcome of rotator cuff repair. One hundred and twenty-five patients who underwent unilateral primary repair of a chronic rotator cuff tear were included in the study. All operations were performed by a single surgeon. Each patient prospectively completed the Simple Shoulder Test (SST); the Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire; three visual analogue scales for shoulder pain, shoulder function, and quality of life; and the Short Form-36 (SF-36) preoperatively and at one year (mean and standard deviation, 54.1 +/- 7.6 weeks) postoperatively. Preoperative expectations were quantified with use of six questions from the Musculoskeletal Outcomes Data Evaluation and Management System (MODEMS) questionnaire. Greater preoperative expectations correlated with better postoperative performance on the SST, DASH, each visual analogue scale, and the SF-36 (p values ranging from <0.0001 to 0.03) as well as with greater improvement from the baseline scores on the DASH and SF-36 (p values ranging from <0.0001 to 0.018). A rigorous multivariate analysis controlling for age, gender, smoking, Workers' Compensation status, symptom duration, number of previous operations, number of comorbidities, tear size, and repair technique confirmed that greater expectations were a significant independent predictor of both better performance at one year and greater improvement on the SST, the DASH, each visual analogue scale, and the SF-36 (p values ranging from <0.001 to 0.042). Patients' preoperative expectations regarding rotator cuff repair are associated with their actual self-assessed outcome. Variations in patient expectations may help to explain divergent results in published series as well as among various patient populations.


Weak Organic Acids Decrease Borrelia burgdorferi Cytoplasmic pH, Eliciting an Acid Stress Response and Impacting RpoN- and RpoS-Dependent Gene Expression.
The spirochete Borrelia burgdorferi survives in its tick vector, Ixodes scapularis, or within various hosts. To transition between and survive in these distinct niches, B. burgdorferi changes its gene expression in response to environmental cues, both biochemical and physiological. Exposure of B. burgdorferi to weak monocarboxylic organic acids, including those detected in the blood meal of fed ticks, decreased the cytoplasmic pH of B. burgdorferi in vitro. A decrease in the cytoplasmic pH induced the expression of genes encoding enzymes that have been shown to restore pH homeostasis in other bacteria. These include putative coupled proton/cation exchangers, a putative Na+/H+ antiporter, a neutralizing buffer transporter, an amino acid deaminase and a proton exporting vacuolar-type VoV1 ATPase. Data presented in this report suggested that the acid stress response triggered the expression of RpoN- and RpoS-dependent genes including important virulence factors such as outer surface protein C (OspC), BBA66, and some BosR (Borrelia oxidative stress regulator)-dependent genes. Because the expression of virulence factors, like OspC, are so tightly connected by RpoS to general cellular stress responses and cell physiology, it is difficult to separate transmission-promoting conditions in what is clearly a multifactorial and complex regulatory web.


Distinct expression profiles of Notch-1 protein in human solid tumors: Implications for development of targeted therapeutic monoclonal antibodies.
Biological therapies, such as monoclonal antibodies (mAbs) that target tumor-associated antigens have been considered an effective therapeutic approach in oncology. In considering Notch-1 receptor as a potential target, we performed immunohistochemistry on tissue microarrays to determine 1) whether the receptor is overexpressed in tumor cells as compared to their corresponding normal tissues and 2) the clinical significance of its expression levels in human breast, colorectal, lung and prostate cancers. We found that the expression of Notch-1 protein was overexpressed in primary colorectal adenocarcinoma and nonsmall cell lung carcinoma (NSCLC), but not in primary ductal breast carcinoma or prostate adenocarcinoma. Further analysis revealed that higher levels of Notch-1 protein expression were significantly associated with poorer differentiation of breast and prostate tumors. Strikingly, for NSCLC, the expression levels of Notch-1 protein were found to be inversely correlated with tumor differentiation and progression. For colorectal tumors, however, no correlation of Notch-1 protein expression was found with any tumor clinicopathological parameters, in spite of its overexpression in tumor cells. Our data demonstrated the complexity of Notch-1 protein expression in human solid tumors and further supported the notion that the roles of Notch-1 expression in tumorigenesis are highly context-dependent. The findings could provide the basis for development of distinct therapeutic strategies of Notch-1 mAbs for its applications in the treatment of suitable types of human cancers.


Tigecycline Heteroresistance and Resistance Mechanism in Clinical Isolates of Acinetobacter baumannii.
Tigecycline is regarded as a last-resort treatment for multidrug-resistant Acinetobacter baumannii. However, tigecycline resistance in A. baumannii has increased worldwide. In this study, we investigated tigecycline heteroresistance in A. baumannii isolates from South Korea. Antibiotic susceptibility testing was performed on 323 nonduplicated A. baumannii isolates. Among 260 and 37 tigecycline-susceptible and -intermediate-resistant A. baumannii isolates, 146 (56.2%) and 22 (59.5%) isolates were identified as heteroresistant to tigecycline through a disk diffusion assay and population analysis profiling. For selected isolates, an in vitro time-kill assay was performed, and survival rates were measured after preincubation with diverse concentrations of tigecycline. Heteroresistant isolates showed regrowth after 12 h of 2× MIC of tigecycline treatment, and resistant subpopulations were selected by preexposure to tigecycline. Furthermore, genetic alterations in adeABC, adeRS, and rpsJ were assessed, and the relative mRNA expression levels of adeB and adeS were compared. The tigecycline resistance in subpopulations might be due to the insertion of ISAba1 in adeS, leading to the overexpression of the AdeABC efflux pump. However, the tigecycline resistance of subpopulations was not stable during serial passages in antibiotic-free medium. The reversion of tigecycline susceptibility by antibiotic-free passages might occur by additional insertions of ISAba10 in adeR and nucleotide alterations in adeS in some mutants. Tigecycline heteroresistance is prevalent in A. baumannii isolates, which results in treatment failure. Tigecycline resistance is mainly due to the overexpression of the AdeABC efflux pump, which is associated with genetic mutations, but this resistance could be reversed into susceptibility by additional mutations in antibiotic-free environments. IMPORTANCE The evidence that antibiotic heteroresistance is responsible for treatment failure in clinical settings is increasing. Thus, detection and characterization of heteroresistance would be important for appropriate therapeutic guidance to treat bacterial infections. However, data on tigecycline heteroresistance in Gram-negative bacteria is currently limited, although tigecycline is regarded as a last-line antibiotic against infections caused by antibiotic-resistant pathogens. In this study, we investigated the tigecycline heteroresistance in Acinetobacter baumannii, which has been listed by the WHO as a priority for research and development of new antibiotics. We found very high prevalence of tigecycline-heteroresistant A. baumannii clinical isolates, which may result in treatment failure due to the selection of resistant subpopulations. We also identified the main resistance mechanism in tigecycline-resistant subpopulations, that is, upregulation of AdeABC efflux pumps due to ISAba1 insertion in adeS.


An EPR, ENDOR and EIE study of gamma-irradiated poly (lactide-co-glycolide) polymers.
Gamma radiation of poly (lactide-co-glycolide) raw polymers and processed microspheres under vacuum and at 77 K results in the formation of a series of free radicals. The resulting powder electron paramagnetic resonance (EPR) spectrum contains a distribution of several different radicals, depending on the annealing temperature, and is therefore difficult to interpret. By utilising the selectivity of the electron nuclear DOuble resonance (ENDOR) and associated ENDOR induced EPR (EIE) techniques, a more direct approach for the deconvolution of the EPR spectrum can be achieved. Using this approach, the radiolytically induced CH3 *CHC(O)R- chain scission radical was identified at 120 K by simulation of the EIE spectrum. At elevated temperatures (250 K), this radical decays considerably and the more stable radicals -O*CHC(O)-, CH3 *C(OR)C(O)- and CH3 *C(OH)C(O)- predominate. This work demonstrates the utility of the EIE approach to supplement and aid the interpretation of powder EPR spectra of radicals in a polymer matrix.


A novel rhabdovirus detected in Anisakis larvae distributed in the coastal areas of Japan: Viral genome analysis and possible coevolutionary relationship between virus and host nematodes.
In the last decade, it has become evident that various RNA viruses infect helminths including Order Ascaridida. However, there is still no information available for viruses infecting Anisakis. We herewith demonstrate the presence of a novel rhabdovirus from Anisakis larvae detected by next-generation sequencing analysis and following RT-PCR. We determined the nearly all nucleotide sequence (12,376 nucleotides) of the viral genome composed of seven open reading frames, and we designated the virus as Suzukana rhabdo-like virus (SkRV). BLASTx search indicated that SkRV is a novel virus belonging to the subfamily Betanemrhavirus, rhabdovirus infecting parasitic nematodes of the Order Ascaridida. SkRV sequence was detectable only in the total RNA but not in the genomic DNA of Anisakis, ruling out the possibility of SkRV being an endogenous viral element incorporated into the host genomic DNA. When we individually tested Anisakis larvae obtained from Scomber japonicus migrating in the coastal waters of Japan, not all but around 40% were SkRV-positive. In the phylogenetic trees of Betanemrhavirus and of the host Ascaridida nematodes, we observed that evolutional distances of viruses were, to some extent, parallel with that of host nematodes, suggesting that viral evolution could have been correlated with evolution of the host. Although biological significance of SkRV on Anisakis larvae is still remained unknown, it is interesting if SkRV were somehow related to the pathogenesis of anisakiasis, because it is important matter of public health in Japan and European countries consuming raw marine fishes.


Vector-host interactions in avian nests: do mosquitoes prefer nestlings over adults?
The hypothesis that nestlings are a significant driver of arbovirus transmission and amplification is based upon findings that suggest nestlings are highly susceptible to being fed upon by vector mosquitoes and to viral infection and replication. Several previous studies have suggested that nestlings are preferentially fed upon relative to adults in the nest, and other studies have reported a preference for adults over nestlings. We directly tested the feeding preference of nestling and adult birds in a natural setting, introducing mosquitoes into nesting boxes containing eastern bluebirds (Sialia sialis), collecting blood-fed mosquitoes, and matching the source of mosquito blood meals to individual birds using microsatellite markers. Neither nestlings nor adults were fed upon to an extent significantly greater than would be predicted based upon their relative abundance in the nests, although feeding upon mothers decreased as the age of the nestlings increased.


The Modified WHO Analgesic Ladder: Is It Appropriate for Chronic Non-Cancer Pain?
From 1986, the World Health Organization (WHO) analgesic ladder has been used as the simple and valuable pain-relieving guidance in the pharmaceutical pain management, however, with the development of medical history, notions about pain physiology and pain management have already updated. Is the analgesic ladder still appropriate for chronic non-cancer pain (CNCP) patients? This study aims to analyse the current usage of the analgesic ladder in patients with CNCP by evaluating previously published pertinent studies. Literature published in English from January 1980 to April 2019 and cited on PubMed database was included. Analysis on the analgesic ladder, current status of CNCP management, and a new revised ladder model were developed based on relevant literature. The WHO analgesic ladder for cancer pain is not appropriate for current CNCP management. It is revised into a four-step ladder: the integrative therapies being adopted at each step for reducing or even stopping the use of opioid analgesics; interventional therapies being considered as step 3 before upgrading to strong opioids if non-opioids and weak opioids failed in CNCP management. A simple and valuable guideline in past years, the WHO analgesic ladder is inappropriate for the current use of CNCP control. A revised four-step analgesic ladder aligned with integrative medicine principles and minimally invasive interventions is recommended for control of CNCP.


Nonhypertensive White Matter Hyperintensities in Stroke: Risk Factors, Neuroimaging Characteristics, and Prognosis.
This study explored the risk factors, neuroimaging features, and prognostic implications of nonhypertensive white matter hyperintensity (WMH) in patients with acute ischemic stroke and transient ischemic attack. We included 2283 patients with hypertension and 1003 without from a pool of 10 602. Associations of moderate-to-severe WMH with known risk factors, functional outcome, and a composite of recurrent stroke, myocardial infarction, and all-cause mortality were evaluated. A subset of 351 patients without hypertension and age- and sex-matched pairs with hypertension and moderate-to-severe WMH was created for a detailed topographic examination of WMH, lacunes, and microbleeds. Approximately 35% of patients without hypertension and 65% of patients with hypertensive stroke exhibited moderate-to-severe WMH. WMH was associated with age, female sex, and previous stroke, irrespective of hypertension. In patients without hypertension, WMH was associated with initial systolic blood pressure and was more common in the anterior temporal region. In patients with hypertension, WMH was associated with small vessel occlusion as a stroke mechanism and was more frequent in the periventricular region near the posterior horn of the lateral ventricle. The higher prevalence of occipital microbleeds in patients without hypertension and deep subcortical lacunes in patients with hypertension were also observed. Associations of moderate-to-severe WMH with 3-month functional outcome and 1-year cumulative incidence of the composite outcome were significant (both P<0.01), although the latter lost significance after adjustments. The associations between WMH and outcomes were consistent across hypertensive status. One-third of patients without hypertension with stroke have moderate-to-severe WMH. The pathogenesis of WMH may differ between patients without and with hypertension, but its impact on outcome appears similar.


Cushing's syndrome.
Cushing's syndrome remains one of the great diagnostic challenges to internists and endocrinologists. Urologists have the primary role in the treatment of many of the forms of Cushing's syndrome, especially adrenal tumors and micronodular hyperplasia. As we continue to gain experience with Cushing's disease treated with pituitary surgery and with etopic ACTH syndrome and the other forms of Cushing's syndrome, urologists are likely to become even more involved in the therapy of all types of Cushing's syndrome.


Incidence and significance of Delphian node metastasis in papillary thyroid cancer.
The objective of this study was to determine the incidence and significance of Delphian node (DN) metastasis in papillary thyroid cancer. Despite the historic association, the prognostic implications of thyroid cancer metastasis to the DN remain unclear. Retrospective analysis of 101 patients who underwent thyroid surgery from 2007 to 2009 for papillary thyroid cancer, and had their DN harvested ab initio. Of these, 25 had metastatic disease to the DN. DN status was correlated with clinical and pathologic factors including age, gender, tumor size, histologic variant, extra-thyroidal extension (ETE), and central and lateral nodal metastasis. DN positivity was seen consequent to higher rates of ETE of the primary tumor (52% vs. 28%; P = 0.025) and is associated with further nodal metastases to the central (72% vs. 22%; P < 0.0001) and lateral (28% vs. 4%; P < 0.0001) neck compartments. DN positivity was also associated with heavier nodal burden, in terms of number of metastatic nodes (median 5 vs. 2 nodes; P = 0.005) and nodal size (median 1 cm vs. 0.5 cm; P = 0.03). Metastatic involvement of the DN is often associated with ETE and increased incidence of metastatic disease to the central and lateral neck compartments. If the DN is positive on intraoperative frozen section, careful evaluation of the central and lateral nodal compartments is essential.


The expression of glucose transporters and mitochondrial division and fusion proteins in rats exposed to hypoxic preconditioning to attenuate propofol neurotoxicity.
Purpose: Evidence has shown that propofol may cause widespread apoptotic neurodegeneration. Hypoxic preconditioning has been demonstrated to provide neuroprotection and brain recovery from both acute and chronic neurodegeneration in several cellular and animal models. However, the mechanism has not been well elucidated. Therefore, the present study was designed to investigate the expression of glucose transporters (GLUT1 and GLUT3) and mitochondrial division and fusion (Drp1 and Mfn2) proteins in rats exposed to hypoxic preconditioning to attenuate propofol neurotoxicity.Methods: Propofol (100 mg/kg) was given to 7-day-old Sprague-Dawley rats; in some rats, hypoxic preconditioning was administered before intraperitoneal propofol injection by subjecting rats to five cycles of 10 min of hypoxia (8% O2) and 10 min of normoxia (21% O2). Then, the rats were allowed to breathe room air for 2 h. Neuronal mitochondrial morphology was observed by transmission electron microscopy. ATP content was detected using an ATP assay kit. The expression levels of GLUT1, GLUT3, pDrp1, Drp1 and Mfn2 were detected by Western blot, and the expression levels of GLUT1 and GLUT3 were further examined by immunohistochemistry.Results: Propofol damaged mitochondria, and decreased ATP content and GLUT3 and pDrp1 protein expression. However, our results suggested that hypoxic preconditioning could attenuate propofol neurotoxicity by reducing mitochondrial damage and increasing ATP content and pDrp1, GLUT1 and GLUT3 protein expression.Conclusion: Hypoxic preconditioning reduced propofol-induced damage in the hippocampus of neonatal rats by attenuating the increase in mitochondrial division and decrease in GLUT3 expression.


Increased food intake leads to obesity and insulin resistance in the tg2576 Alzheimer's disease mouse model.
Recent studies suggest that hyperinsulinemia and insulin resistance are linked to Alzheimer's disease (AD). In this study, we used Tg2576 transgenic (Tg) mice, a widely used transgenic mouse model for AD, to explore the relationship between increased amyloid beta-peptide (Abeta) and insulin resistance. When fed a high-fat diet (HFD), Tg mice developed obesity and insulin resistance at 16 wk of age. Furthermore, HFD-fed Tg mice displayed abnormal feeding behavior and increased caloric intake with time. Although caloric intake of HFD-fed Tg mice was similar to that of normal diet-fed Tg or wild-type mice during 4 to 8 wk of age, it increased sharply at 12 wk, and went up further at 16 wk, which paralleled changes in the level of Abeta40 and Abeta42 in the brain of these mice. Limiting food intake in HFD-fed Tg mice by pair-feeding a caloric intake identical with that of normal diet-fed mice completely prevented the obesity and insulin intolerance of HFD-fed Tg mice. The hypothalamus of HFD-fed Tg mice had a significant decrease in the expression of the anorexigenic neuropeptide, brain-derived neurotrophic factor, at both the mRNA and protein levels. These findings suggest that the increased Abeta in the brain of HFD-fed Tg2576 mice is associated with reduced brain-derived neurotrophic factor expression, which led to abnormal feeding behavior and increased food intake, resulting in obesity and insulin resistance in these animals.


Calcitonin gene-related peptide elevates cyclic AMP levels in chick skeletal muscle: possible neurotrophic role for a coexisting neuronal messenger.
Recent immunocytochemical studies have shown that calcitonin gene-related peptide (CGRP) coexists with the neurotransmitter acetylcholine in spinal motoneurons of the chick. Moreover, CGRP causes an increase in the number of acetylcholine receptors on the surface of cultured chick myotubes. CGRP might thus serve as one of the signals by which motoneurons regulate endplate development. In a search for the second messengers involved, we now demonstrate that CGRP stimulates accumulation of cyclic AMP (cAMP) in cultured chick myotubes. This effect is, at least in part, mediated by an increase in cAMP synthesis, as the peptide also activates adenylate cyclase in chick muscle membranes. Nanomolar concentrations of CGRP elicit significant increases in both cellular cAMP levels and acetylcholine receptor numbers. The present findings suggest that cAMP is one of the second messengers which mediate the increase in acetylcholine receptor number elicited by CGRP. Furthermore, CGRP might be implicated in other trophic actions mediated by cAMP in skeletal muscle cells.


Linking the Tinnitus Questionnaire and the subjective Clinical Global Impression: which differences are clinically important?
Development of new tinnitus treatments requires prospective placebo-controlled randomized trials to prove their efficacy. The Tinnitus Questionnaire (TQ) is a validated and commonly used instrument for assessment of tinnitus severity and has been used in many clinical studies. Defining the Minimal Clinically Important Difference (MCID) for TQ changes is an important step to a better interpretation of the clinical relevance of changes observed in clinical trials. In this study we aimed to estimate the minimum change of the TQ score that could be considered clinically relevant. 757 patients with chronic tinnitus were pooled from the TRI database and the RESET study. An anchor-based approach using the Clinical Global Impression (CGI) scale and distributional approaches were used to estimate MCID. Receiver Operating Characteristic (ROC) curves were calculated to define optimal TQ change cutoffs discriminating between minimally changed and unchanged subjects. The relationship between TQ change scores and CGI ratings of change was good (r = 0.52, p < 0.05). Mean change scores associated with minimally better and minimally worse CGI categories were -6.65 and +2.72 respectively. According to the ROC method MCID for improvement was -5 points and for deterioration +1 points. Distribution and anchor-based methods yielded comparable results in identifying MCIDs. ΔTQ scores of -5 and +1 points were identified as the minimal clinically relevant change for improvement and worsening respectively. The asymmetry of the MCIDs for improvement and worsening may be related to expectation effects.


Intramammary lymph node metastasis in a patient with ovarian carcinoma and a brief review of the literature.
Metastasis from serous carcinoma of the ovary usually occurs in the subdiaphragmatic region. Metastasis to the breast and/or axillary-intramammary lymph node is very rare. It usually occurs in advanced disease, and it is important to distinguish a primary breast cancer from an ovarian cancer metastasis since the management of these two entities is totally different. Here we present a patient with metastasis to the breast and intramammary lymph nodes from ovarian serous carcinoma 25 months after diagnosis.


Sustainable Valorization of Sambucus nigra L. Berries: From Crop Biodiversity to Nutritional Value of Juice and Pomace.
Improvement of dietary and ecological biodiversity, namely by exploring autochthonous varieties, is a key point to the construction of a more sustainable food system and planetary health. However, the environmental sustainability continues to face huge challenges, reflecting the importance of achieving a better understanding about the functional role of biodiversity in ecosystems. Thus, the main objective of this research is to contribute to the sustainable valorization of Sambucus nigra L. berries through a comprehensive approach to evaluate the effects of elderberry's cultivar, harvest year, and plantation field on the physicochemical berry composition. Moreover, the nutritional value of elderberry juice and respective dried pomace was determined. This complementary information is of huge utility for the rational and, as much as possible, integral use of elderberries. The harvest year, followed by field and the interaction of harvest × field, accounted for the highest impact on the berry's physicochemical parameters, indicating the importance of the combined impact of the macro- and mesoclimate conditions on plant metabolism. Elderberry juice and dried pomace are a good source of carbohydrates (ca. 12 and 82%, respectively) and have low amounts of fat (≤2.5%), making them low-energy foods. Dried pomace may also represent a potential alternative source of vegetal protein (ca. 6%).


Energy costs of blood digestion in a host-specific haematophagous parasite.
Fleas consume and digest blood from their hosts. We hypothesized that the energy costs of digestion of blood by fleas is dependent on the host species. To test this hypothesis, we studied CO2 emission, a measure of energy expenditure, during digestion of a blood meal taken by Parapulex chephrenis from a preferred (Acomys cahirinus) and a non-preferred (Gerbillus dasyurus) host. We predicted that the energy cost of digestion would be lower for A. cahirinus blood than that for G. dasyurus. Male and female fleas consumed similar amounts of blood per unit body mass, independent of host species. Our prediction was supported in that fleas expended significantly more energy digesting blood of G. dasyurus than blood of A. cahirinus. We also found CO2 emission rates of fed fleas were higher than those of unfed fleas and differed significantly among stages of blood digestion when a flea fed on G. dasyurus but not when it fed on A. cahirinus. When fed on G. dasyurus, fleas spent less energy during earlier than later stages of digestion.


Ezrin: a regulator of actin microfilaments in cell junctions of the rat testis.
Ezrin, radixin, moesin and merlin (ERM) proteins are highly homologous actin-binding proteins that share extensive sequence similarity with each other. These proteins tether integral membrane proteins and their cytoplasmic peripheral proteins (e.g., adaptors, nonreceptor protein kinases and phosphatases) to the microfilaments of actin-based cytoskeleton. Thus, these proteins are crucial to confer integrity of the apical membrane domain and its associated junctional complex, namely the tight junction and the adherens junction. Since ectoplasmic specialization (ES) is an F-actin-rich testis-specific anchoring junction-a highly dynamic ultrastructure in the seminiferous epithelium due to continuous transport of germ cells, in particular spermatids, across the epithelium during the epithelial cycle-it is conceivable that ERM proteins are playing an active role in these events. Although these proteins were first reported almost 25 years and have since been extensively studied in multiple epithelia/endothelia, few reports are found in the literature to examine their role in the actin filament bundles at the ES. Studies have shown that ezrin is also a constituent protein of the actin-based tunneling nanotubes (TNT) also known as intercellular bridges, which are transient cytoplasmic tubular ultrastructures that transport signals, molecules and even organelles between adjacent and distant cells in an epithelium to coordinate cell events that occur across an epithelium. Herein, we critically evaluate recent data on ERM in light of recent findings in the field in particular ezrin regarding its role in actin dynamics at the ES in the testis, illustrating additional studies are warranted to examine its physiological significance in spermatogenesis.


Platelet-endothelial associations may promote cytomegalovirus replication in the salivary gland in mice.
Platelet decline is a feature of many acute viral infections, including cytomegalovirus (CMV) infection in humans and mice. Platelet sequestration in association with other cells, including endothelium and circulating leukocytes, can contribute to this decline and influence the immune response to and pathogenesis of viral infection. We sought to determine if platelet-endothelial associations (PEAs) contribute to platelet decline during acute murine CMV (mCMV) infection, and if these associations affect viral load and production. Male BALB/c mice were infected with mCMV (Smith strain), euthanized at timepoints throughout acute infection and compared to uninfected controls. An increase in PEA formation was confirmed in the salivary gland at all post-inoculation timepoints using immunohistochemistry for CD41+ platelets co-localizing with CD34+ vessels. Platelet depletion did not change amount of viral DNA or timecourse of infection, as measured by qPCR. However, platelet depletion reduced viral titer of mCMV in the salivary glands while undepleted controls demonstrated robust replication in the tissue by plaque assay. Thus, platelet associations with endothelium may enhance the ability of mCMV to replicate within the salivary gland. Further work is needed to determine the mechanisms behind this effect and if pharmacologic inhibition of PEAs may reduce CMV production in acutely infected patients.


Effects of chronic ethanol treatment on the beta-adrenergic receptor-coupled adenylate cyclase system of mouse cerebral cortex.
Chronic ingestion of ethanol, which produced tolerance and physical dependence, resulted in altered function of the cerebral cortical beta-adrenergic receptor-coupled adenylate cyclase system in mice. Although there was no change in basal adenylate cyclase activity, or in the activity of the digitonin-solubilized catalytic unit, stimulation of adenylate cyclase activity by the nonhydrolyzable guanine nucleotide analog guanylylimidodiphosphate [Gpp(NH)p] was reduced in brains of ethanol-fed animals. Ethanol added in vitro increased adenylate cyclase activity, and this enhancement, in the presence of Gpp(NH)p, was also reduced in cortical membranes of ethanol-fed mice. Furthermore, the maximal response to isoproterenol was decreased, and the EC50 for isoproterenol stimulation of adenylate cyclase activity was increased in ethanol-fed animals. The results are consistent with a qualitative or quantitative defect in the function of the stimulatory guanine nucleotide-binding protein (Ns), as well as in the beta-adrenergic receptor, after chronic ethanol exposure. In part, these changes appear to be similar to those that occur during heterologous desensitization of various receptor systems, and may be associated with dependence on or tolerance to ethanol.


Modic Changes and Disc Degeneration of Nonfused Segments 27 to 45 Years After Harrington Instrumentation for Adolescent Idiopathic Scoliosis: Comparison to Healthy Controls.
A case-control study. The aim of this study was to investigate the long-term prevalence of the Modic changes and disc degeneration in the nonfused segments of the surgically treated adolescent idiopathic scoliosis (AIS) patients and compare with age- and sex-matched healthy controls. No studies have looked at the association between Modic changes and low back pain in patients who underwent AIS surgery many years ago. Study subjects consisted of 194 patients with AIS who underwent spinal fusion with Harrington instrumentation between 1968 and 1987. Twenty-six patients (AIS group) underwent lumbar magnetic resonance imaging (MRI), whole spine X-ray, and patient-reported outcomes evaluations [Scoliosis Research Society-22 Patient Questionnaire (SRS-22), Oswestry Disability Index (ODI)]. The mean duration of follow-up observation was 36.1 years. The lowest fusion vertebra was used as follows: T12: 2 patients, L1: 4, L2: 11, L3: 6, L4: 2, L5: 1. Twenty-nine healthy age- and sex-matched individuals were selected as a control (CTR) group. On the basis of MRI findings, a significantly higher percentage of subjects showed Modic changes in the AIS group (AIS group: 57.7%, CTR group: 13.8%). There were no significant differences in the percentage of subjects with disc degeneration rated Pfirrmann grade 4 or higher (AIS group: 61.5%, CTR group: 65.5%). SRS-22 scores for function and self-image were significantly lower in the AIS group. ODI was significantly worse in the AIS group. Modic changes in AIS group were found in the concave side of the curve in 61.9%. The magnitude of lumbar coronal curve was found to be a significant risk factor of Modic changes. Modic changes were observed in 57.7% of AIS patients 27 years or more after spinal fusion with Harrington instrumentation. The magnitude of lumbar coronal curve was found to be a significant risk factor of Modic changes. Modic change occurred at a higher frequency in AIS patients than CTR individuals, with lower function and worse ODI scores. 4.


Fabrication of three-dimensional nano, micro and micro/nano scaffolds of porous poly(lactic acid) by electrospinning and comparison of cell infiltration by Z-stacking/three-dimensional projection technique.
The use of electrospun extracellular matrix (ECM)-mimicking nanofibrous scaffolds for tissue engineering is limited by poor cellular infiltration. The authors hypothesised that cell penetration could be enhanced in scaffolds by using a hierarchical structure where nano fibres are combined with micron-scale fibres while preserving the overall scaffold architecture. To assess this, we fabricated electrospun porous poly(lactic acid) (PLA) scaffolds having nanoscale, microscale and combined micro/nano architecture and evaluated the structural characteristics and biological response in detail. Although the bioactivity was intermediate to that for nanofibre and microfibre scaffold, a unique result of this study was that the micro/nano combined fibrous scaffold showed improved cell infiltration and distribution than the nanofibrous scaffold. Although the cells were found to be lining the scaffold periphery in the case of nanofibrous scaffold, micro/nano scaffolds had cells dispersed throughout the scaffold. Further, as expected, the addition of nanoparticles of hydroxyapatite (nHAp) improved the bioactivity, although it did not play a significant role in cell penetration. Thus, this strategy of creating a three-dimensional (3D) micro/nano architecture that would increase the porosity of the fibrous scaffold and thereby improving the cell penetration, can be utilised for the generation of functional tissue engineered constructs in vitro.


Progressive myoclonic epilepsy: myoclonic epilepsy and ataxia due to KCNC1 mutation (MEAK): a case report and review of the literature.
Progressive myoclonic epilepsy (PME) is characterized by prominent myoclonus and generalized or focal seizures. A recently described novel KCNC1 mutation is associated with a specific phenotype of progressive myoclonic epilepsy, which has been defined as myoclonic epilepsy and ataxia due to potassium channel mutation (MEAK). Our case illustrates a typical presentation of this disease and the potential for misdiagnosis as idiopathic generalized epilepsy during the early phase of the disease. Unique findings that may suggest an alternative diagnosis are a progressive myoclonus, prominent ataxia/dysmetria on examination, and abnormally high amplitude in the sensory evoked potential recording. We also report a brief review of the existing literature on MEAK. Early and accurate diagnosis with genetic testing may significantly help in counseling patients and families.


The genetic variation in the R1a clade among the Ashkenazi Levites' Y chromosome.
Approximately 300,000 men around the globe self-identify as Ashkenazi Levites, of whom two thirds were previously shown to descend from a single male. The paucity of whole Y-chromosome sequences precluded conclusive identification of this ancestor's age, geographic origin and migration patterns. Here, we report the variation of 486 Y-chromosomes within the Ashkenazi and non-Ashkenazi Levite R1a clade, other Ashkenazi Jewish paternal lineages, as well as non-Levite Jewish and non-Jewish R1a samples. Cumulatively, the emerging profile is of a Middle Eastern ancestor, self-affiliating as Levite, and carrying the highly resolved R1a-Y2619 lineage, which was likely a minor haplogroup among the Hebrews. A star-like phylogeny, coalescing similarly to other Ashkenazi paternal lineages, ~1,743 ybp, suggests it to be one of the Ashkenazi paternal founders; to have expanded as part of the overall Ashkenazi demographic expansion, without special relation to the Levite affiliation; and to have subsequently spread to non-Ashkenazi Levites.


Auditory function of Friedreich's ataxia. Electrophysiologic study of a family.
Hearing function and auditory brainstem potentials were evaluated in nine members of three generations of a family affected with Friedreich's ataxia. The characteristic findings were high-frequency sensorineural hearing impairment of varying severity and diminished or absent N1, N2, and N3 responses. These electrophysiologic aberrations correlate topographically with previously reported pathological observations.


Effect of Chemical Degradation of Sodium Alginate on Capsaicin Encapsulation.
Capsaicin is known as an oily extract of paprika that is characterized by pungent taste and bioactivity. It also may cause irritation to the mouth and stomach which is why is so important to immobilize capsaicin on a carrier to prevent it. The usage of alginate oligomers, which has an antioxidant potential compared to alginate, is of benefit because it may be used in the immobilization of bioactive substances that are fragile to oxidation. The purpose of this study was to use sodium alginate oligomers as a coating material in the encapsulation process of paprika oleoresin. Alginate oligomers were produced by chemical degradation with hydrogen peroxide. The characteristics of the samples were obtained by measuring the viscosity, the contact angle of the surface, and the surface tension of solutions. The obtained solution of alginate oligomers served as the carrier material for the immobilization of capsaicin. Capsules were prepared by ionic gelation using a calcium chloride solution as a crosslinking agent. In this way, capsules without and with the core (capsaicin) were prepared and their ability to scavenge free radicals (DPPH) and iron-reducing properties (FRAP) were determined. The stability of the capsules was examined by thermal decomposition and under conditions of the gastric and small intestine, and capsaicin content was detected using high-performance liquid chromatography. It was found that alginate oligomers could be used in the encapsulation of bioactive compounds and the efficiency was above 80%. Capsule production from alginate oligomers affected their thermal stability. The use of alginate derivatives as a carrier increased the antioxidant properties of the finished product, as well as its ability to reduce iron ions. The use of alginate oligomers as a coating material prevented the active substance from being released too early in the conditions of the small intestine, prolonged the stability of the capsules, and supported their durability in gastric conditions.


Deciphering sulfoglycolipids of Mycobacterium tuberculosis.
For 4 decades, in vivo and in vitro studies have suggested that sulfoglycolipids (SGLs) play a role in the virulence or pathogenesis of the tubercle bacilli. However, the SGL structure and biosynthesis pathway remain only partially elucidated. Using the modern tools of structural analysis, including MALDI-time-of-flight MS, MS/MS, and two-dimensional NMR, we reevaluated the structure of the different SGL acyl (di-, tri-, and tetra-acylated) forms of the reference strain Mycobacterium tuberculosis H37Rv, as well as those produced by the mmpL8 knockout strains previously described to intracellularly accumulate di-acylated SGL. We report here the identification of new acyl forms: di-acylated SGL esterified by simple fatty acids only, as well as mono-acylated SGL bearing a hydroxyphthioceranoic acid, which were characterized in the wild-type strain. In a clinical strain, a complete family of mono-acylated SGLs was characterized in high abundance for the first time. For the mmpL8 mutant, SGLs were found to be esterified i) by an oxophthioceranoic acid, never observed so far, and ii) at nonconventional positions in the case of the unexpected tri-acylated forms. Our results further confirm the requirement of MmpL8 for the complete assembly of the tetra-acylated forms of SGL and also provide, by the discovery of new intermediates, insights in terms of the possible SGL biosynthetic pathways.


Invasive alien plants benefit more from clonal integration in heterogeneous environments than natives.
What confers invasive alien plants a competitive advantage over native plants remains open to debate. Many of the world's worst invasive alien plants are clonal and able to share resources within clones (clonal integration), particularly in heterogeneous environments. Here, we tested the hypothesis that clonal integration benefits invasive clonal plants more than natives and thus confers invasives a competitive advantage. We selected five congeneric and naturally co-occurring pairs of invasive alien and native clonal plants in China, and grew pairs of connected and disconnected ramets under heterogeneous light, soil nutrient and water conditions that are commonly encountered by alien plants during their invasion into new areas. Clonal integration increased biomass of all plants in all three heterogeneous resource environments. However, invasive plants benefited more from clonal integration than natives. Consequently, invasive plants produced more biomass than natives. Our results indicate that clonal integration may confer invasive alien clonal plants a competitive advantage over natives. Therefore, differences in the ability of clonal integration could potentially explain, at least partly, the invasion success of alien clonal plants in areas where resources are heterogeneously distributed.


Photoluminescent and electrochemiluminescent dual-signaling probe for bio-thiols based on a ruthenium(II) complex.
Photoluminescence (PL) and electrochemiluminescence (ECL) detection techniques are highly sensitive and widely used methods for clinical diagnostics and analytical biotechnology. In this work, a unique ruthenium(II) complex, [Ru(bpy)(2)(DNBSO-bpy)](PF(6))(2) (bpy: 2,2'-bipyridine; DNBSO-bpy: 2,4-dinitrobenzenesulfonate of 4-(4-hydroxyphenyl)-2,2'-bipyridine), has been designed and synthesized as a highly sensitive and selective PL and ECL dual-signaling probe for the recognition and detection of bio-thiols in aqueous media. As a thiol-responsive probe, the complex can specifically and rapidly react with bio-thiols in aqueous solutions to yield a bipyridine-Ru(II) complex derivative, [Ru(bpy)(2)(HP-bpy)](2+) (HP-bpy: 4-(4-hydroxyphenyl)-2,2'-bipyridine), accompanied by the remarkable PL and ECL enhancements. The complex was used as a probe for the PL and ECL detections of cysteine (Cys) and glutathione (GSH) in aqueous solutions. The dose-dependent PL and ECL enhancements showed good linear relationships against the Cys/GSH concentrations with the detection limits at nano-molar concentration level. Moreover, the complex-loaded HeLa cells were prepared for PL imaging of the endogenous intracellular thiols. The results demonstrated the practical utility of the complex as a cell-membrane permeable probe for PL imaging detection of bio-thiols in living cells.


Molecular and functional characterization of the p62 complex, an assembly of nuclear pore complex glycoproteins.
Macromolecular trafficking across the nuclear envelope involves interactions between cytosolic transport factors and nuclear pore complex proteins. The p62 complex, an assembly of 62, 58, 54, and 45-kD O-linked glycoproteins-localized near the central gated channel of the nuclear pore complex, has been directly implicated in nuclear protein import. The cDNA cloning of rat p62 was reported previously. We have now carried out cDNA cloning of rat p58, p54, and p45. We found that p58 contains regions with FG (Phe, Gly) and PA (Pro, Ala) repeats at both its NH2 and COOH termini separated by a predicted alpha-helical coiled-coil region, while p54 has an NH2-terminal FG and PA repeat region and a COOH-terminal predicted coiled-coil region. p45 and p58 appear to be generated by alternative splicing, with p45 containing the NH2-terminal FG repeat region and the coiled-coil region of p58. Using immunogold electron microscopy, we found that p58/p45 and p54 are localized on both sides of the nuclear pore complex, like p62. Previous studies have shown that immobilized recombinant p62 can bind the cytosolic nuclear import factor NTF2 and thereby deplete transport activity from cytosol. We have now found that immobilized recombinant p58 and p54 also can deplete nuclear transport activity from cytosol, and that p62, p58, and p54 bind directly to the cytosolic nuclear import factors p97 and NTF2. At least in the case of p58, this involves FG repeat regions. Moreover, p58 can bind to a complex containing transport ligand, the nuclear localization sequence receptor (Srp1 alpha) and p97. These data support a model in which the p62 complex binds to a multicomponent particle consisting of transport ligand and cytosolic factors to achieve accumulation of ligand near the central gated channel of the nuclear pore complex.


Long-Term Mean Vertical Motion over the Tropical Pacific: Wind-Profiling Doppler Radar Measurements.
Measurement from Christmas Island (2 degrees N, 157 degrees W) of long-term mean vertical motions in the tropical atmosphere using very-high-frequency wind-profiling Doppler radar show that there is a transition from downward motion in the free troposphere to upward motion in the upper troposphere and lower stratosphere. The observations in the free troposphere are consistent with a balance between adiabatic and diabatic heating and cooling rates in a clear atmosphere. Comparison of the results at Christmas Island during El Niño and non-El Niño conditions with earlier results obtained for stratiform rain conditions over Pohnpei, Federated States of Micronesia, show that cirrus clouds in the vicinity of the tropopause likely play an important role in determining the sense and magnitude of vertical motions in this region. These results have implications for the exchange of mass between the troposphere and stratosphere over the tropics.


Protein disaggregation machineries in the human cytosol.
Proteins carry out the vast majority of functions in cells, but can only do so when properly folded. Following stress or mutation, proteins can lose their proper fold, resulting in misfolding, inactivity, and aggregation-posing a threat to cellular health. In order to counteract protein aggregation, cells have evolved a remarkable subset of molecular chaperones, called protein disaggregases, which collaboratively possess the ability to forcibly untangle protein aggregates. Here, we review the different chaperone disaggregation machineries present in the human cytosol and their mechanisms of action. Understanding, how these disaggregases function, is both universally and clinically important, as protein aggregation has been linked to multiple, debilitating neurodegenerative diseases.


The Epstein-Barr Virus Regulome in Lymphoblastoid Cells.
Epstein-Barr virus (EBV) transforms B cells to continuously proliferating lymphoblastoid cell lines (LCLs), which represent an experimental model for EBV-associated cancers. EBV nuclear antigens (EBNAs) and LMP1 are EBV transcriptional regulators that are essential for LCL establishment, proliferation, and survival. Starting with the 3D genome organization map of LCL, we constructed a comprehensive EBV regulome encompassing 1,992 viral/cellular genes and enhancers. Approximately 30% of genes essential for LCL growth were linked to EBV enhancers. Deleting EBNA2 sites significantly reduced their target gene expression. Additional EBV super-enhancer (ESE) targets included MCL1, IRF4, and EBF. MYC ESE looping to the transcriptional stat site of MYC was dependent on EBNAs. Deleting MYC ESEs greatly reduced MYC expression and LCL growth. EBNA3A/3C altered CDKN2A/B spatial organization to suppress senescence. EZH2 inhibition decreased the looping at the CDKN2A/B loci and reduced LCL growth. This study provides a comprehensive view of the spatial organization of chromatin during EBV-driven cellular transformation.


In vitro study of patient's and physician's radiation exposure in the performance of epiduroscopy.
Epiduroscopy is a minimally invasive diagnostic and therapeutic technique, useful in the management of patients with back and leg pain. However, the dose of radiation exposure by fluoroscopy during epiduroscopy is not known. The endpoint of our study was to evaluate the amount of radiation exposure for patients and health care workers during epiduroscopy. First, we measured the radiation dose during a 10-minute fluoroscopy exposure in humanoid models, which substituted for the patient and the physician. Second, we measured the duration of fluoroscopy during our clinical epiduroscopy in 14 patients and observed for radiation injury in these patients. In the humanoid models, the patient model skin exposure dose over a 10-minute period was measured as 238 mGy. The physician's exposure dose for 10 minutes was measured as 0.67 mGy outside the lead apron and 0.0084 mGy inside the lead apron. For the clinical epiduroscopic procedures, the average duration of fluoroscopy was 9 minutes and 26 seconds. No skin injuries in the patients were observed at a 1-month postprocedure assessment. The radiological dosages in the patient humanoid model were less than the threshold doses that could lead to organ injuries for 1 epiduroscopic procedure. However, care should be taken for cumulative exposures in repeated procedures.


Albumin-mediated incorporation of water-insoluble therapeutics in layer-by-layer assembled thin films and microcapsules.
The present study demonstrates a method to deliver hydrophobic drugs by incorporation into thin films and microcapsules fabricated via a layer-by-layer assembly approach. The hydrophobic molecule binding properties of albumin have been exploited for solubilization of a water-insoluble molecule, pyrene (model drug), by preparation of non-covalent conjugates with bovine serum albumin (BSA). Conjugation with BSA renders a highly negative zeta potential to the previously uncharged pyrene which favors the assembly formation by electrostatic interaction with a positively charged polyelectrolyte, chitosan (at acidic pH). The growth of the assembly was followed by monitoring pyrene absorbance with successive layer deposition. The thin film assembly was demonstrated to be capable of releasing its hydrophobic cargo under physiological conditions. We demonstrated the applicability of this approach by encapsulating a water-insoluble drug, curcumin. These assemblies were further loaded with the anti-cancer drug Doxorubicin. Biocompatible calcium carbonate microparticles were used for capsule preparation. The porous nature of the microparticles allows for the pre-encapsulation of therapeutic macromolecules like protein. The fabrication of protein encapsulated stable microcapsules with hydrophobic molecules incorporated into the shell of the microcapsules has been demonstrated. The microcapsules were further capable of loading hydrophilic molecules like Rhodamine B. Thus, using the approach described, a multi-agent carrier for hydrophobic and hydrophilic drugs as well as therapeutic macromolecules can be envisioned.


Mitochondrial function in oncogene-transfected rat fibroblasts isolated from multicellular spheroids.
Two mitochondrion-specific fluorochromes, 10-N-nonyl acridine orange (NAO) and rhodamine 123 (Rh123), were used to determine the mechanism responsible for alterations in energy metabolism of transformed rat embryo fibroblast cells isolated from different locations within multicellular spheroids. Accumulation of Rh123 depends on intact mitochondrial membrane potential, whereas NAO is taken up by mitochondria independently of their function and thus represents mitochondrial distribution only. A reproducible selective dissociation procedure was used to isolate cells from different locations within the spheroids. After isolation, cells were simultaneously stained with one mitochondrial stain and the DNA dye Hoechst 33342, and several parameters, including cell volume, were monitored via multilaser-multiparameter flow cytometry. Our data clearly show a decrease in the uptake of Rh123 in cells from the periphery to the inner regions of the tumor spheroids, reflecting a persistent alteration in mitochondrial function. However, NAO staining experiments showed no reduction in the total mitochondrial mass per unit cell volume. Because cells were exposed to stain under uniform conditions after isolation from the spheroid, these data indicate the downregulation of mitochondrial function is associated with cell quiescence rather than a transient effect of reduced nutrient availability. This result, which is in accordance with data from two other cell lines (EMT6 and 9L), might reflect a general phenomenon in multicellular spheroids, supporting the hypothesis that quiescent cells in the innermost viable spheroid layer stably reduce their mitochondrial function, presumably to compensate for lower nutrient supply and/or decreased energy demand.


Impacts of CYP2C19 Polymorphism and Clopidogrel Dosing on in-Stent Restenosis: A Retrospective Cohort Study in Chinese Patients.
This retrospective cohort study is to analyze the impacts of CYP2C19 polymorphism and clopidogrel dosing on in-stent restenosis (ISR) after coronary stenting. Totally, 111 patients were included, who underwent percutaneous coronary intervention (PCI) with drug-eluting stent. Patients received clopidogrel treatment after the intervention on the background treatment with aspirin, based on the genotypes: 75 mg clopidogrel once each day for subjects without CYP2C19 loss-of-function (LOF) alleles (n=51; EM), 75 mg clopidogrel once each day (n=27; IM75) or twice each day (n=33; IM150) for subjects with one CYP2C19 LOF allele. ISR at 3-18 months after coronary stenting was assessed. ISR rate was significantly higher in the IM75 group (40.7%) than the EM group (11.8%). ISR rate in the IM150 group was lower than the IM75 group (6.1% vs 40.7%), and comparable to that in the EM group. Multivariate logistic regression showed that both CYP2C19 genotype and clopidogrel dosing were associated with the risk of ISR after adjusting the relevant confounding factors. ISR risk was higher in the IM patients than the EM patients. Patients with clopidogrel dose of 75 mg once each day had significantly higher risk of ISR than those with the dose of 75 mg twice each day. Increased dose of clopidogrel may reduce the risk of ISR after PCI in CYP2C19 LOF allele(s) carriers. The presence of CYP2C19 LOF allele(s) increases the risk of ISR after stenting, which could be counteracted by the increased dose of clopidogrel.


Clinical and radiological results after Internal Brace suture versus the all-inside reconstruction technique in anterior cruciate ligament tears 12 to 18 months after index surgery.
Anterior cruciate ligament (ACL) injury can lead to reduced function, meniscal lesions, and early joint degeneration. Preservation of a torn ACL using the Internal Brace technique might re-establish normal knee kinematics, avoid donor-site morbidity due to tendon harvesting, and potentially maintain proprioception of the knee. Fifty subjects were recruited for this study between December 2015 and October 2016. Two groups of individuals who sustained a unilateral ACL rupture were included: those who underwent surgery with preservation of the injured ACL (Internal Brace technique; IB) and those who underwent ACL reconstruction using a hamstring tendon graft (all-inside technique; AI). Subjective self-administered scores were used: the German version of the IKDC Subjective Knee Form (International Knee Documentation Committee), the German version of the WOMAC (Western Ontario and McMaster Universities Arthritis Index), SF-36 (short form), the German version of the KOOS (Knee Osteoarthritis Outcome Score), and the German version of themodified Lysholm Score by Lysholm and Gillquist. Anterior tibial translation was assessed using the KT-1000 Arthrometer (KT-1000 Knee Ligament Arthrometer, MEDmetric Corp., San Diego, CA, USA). Magnetic resonance evaluation was performed in all cases. Twenty-three subjects (46 %) were men, and the mean age was 34.7 years. The objective IKDC scores were "normal" in 15 and 14 patients, "nearly normal" in 11 and 7 patients, and "abnormal" in 1 and 2 patients, in the IB and AI groups, respectively. KT-1000 assessment showed a sideto-side difference of more than 3 mm on maximum manual testing in 11 (44 %) and 6 subjects (28.6 %) in the IB and AI groups, respectively. In the postoperative MRI, 20 (74 %) and 22 subjects (96 %) in the IB and AI groups had an intact ACL. Anterior tibial translation was significantly higher in the IB group compared with the AI group in the manual maximum test. Preservation of the native ACL with the Internal Brace primary repair technique can achieve comparable results to ACL reconstruction using Hamstring autografts over a short term. Clinically relevant limitations such as a higher incidence of pathologic laxity, with patients more prone to pivot-shift phenomenon were observed during the study period.


Extracellular signal-regulated kinase 2 (ERK2) phosphorylation sites and docking domain on the nuclear pore complex protein Tpr cooperatively regulate ERK2-Tpr interaction.
Identifying direct substrates of mitogen-activated protein kinases (MAPKs) and understanding how those substrates are selected is central to understanding how these ubiquitously activated enzymes generate diverse biological responses. In previous work, we identified several new candidate substrates for the MAPK ERK2 (extracellular signal-regulated kinase 2), including the nuclear pore complex protein Tpr (translocated promoter region). In this report, we identify sites on Tpr for ERK2 phosphorylation and binding and demonstrate their functional interaction. ERK2 phosphorylation and dimerization are necessary for ERK2-Tpr binding, and this occurs through a DEF (docking site for ERK2, FXF) domain on Tpr. Surprisingly, the DEF domain and the phosphorylation sites displayed positive cooperativity to promote ERK2 binding to Tpr, in contrast to substrates where phosphorylation reduces binding. Ectopic expression or depletion of Tpr resulted in decreased movement of activated ERK2 from the cytoplasm to the nucleus, implying a role for Tpr in ERK2 translocation. Collectively, the data provide direct evidence that a component of the nuclear pore complex is a bona fide substrate of ERK2 in vivo and that activated ERK2 stably associates with this substrate after phosphorylation, where it could play a continuing role in nuclear pore function. We propose that Tpr is both a substrate and a scaffold for activated ERKs.


T cell protein tyrosine phosphatase attenuates T cell signaling to maintain tolerance in mice.
Many autoimmune diseases exhibit familial aggregation, indicating that they have genetic determinants. Single nucleotide polymorphisms in PTPN2, which encodes T cell protein tyrosine phosphatase (TCPTP), have been linked with the development of several autoimmune diseases, including type 1 diabetes and Crohn's disease. In this study, we have identified TCPTP as a key negative regulator of TCR signaling, which might explain the association of PTPN2 SNPs with autoimmune disease. We found that TCPTP dephosphorylates and inactivates Src family kinases to regulate T cell responses. Using T cell-specific TCPTP-deficient mice, we established that TCPTP attenuates T cell activation and proliferation in vitro and blunts antigen-induced responses in vivo. TCPTP deficiency lowered the in vivo threshold for TCR-dependent CD8(+) T cell proliferation. Consistent with this, T cell-specific TCPTP-deficient mice developed widespread inflammation and autoimmunity that was transferable to wild-type recipient mice by CD8(+) T cells alone. This autoimmunity was associated with increased serum levels of proinflammatory cytokines and anti-nuclear antibodies, T cell infiltrates in non-lymphoid tissues, and liver disease. These data indicate that TCPTP is a critical negative regulator of TCR signaling that sets the threshold for TCR-induced naive T cell responses to prevent autoimmune and inflammatory disorders arising.


Colloid infusions reduce glomerular filtration in resuscitated burn victims.
Colloids are used clinically to minimize edema yet may have detrimental consequences on glomerular filtration. The purpose of this study is to assess the renal and hormonal effects of colloid supplementation in the fluid resuscitation of burn victims. Analytic cohort study. Immediately following their 24 hour post-burn fluid resuscitation with Ringer's lactate, six burn patients (% total body surface area burn 30-57%) were given primed, continuous infusions of inulin and p-aminohippuric acid for 6 hours. Albumin (25% solution, 3 mL/kg/h) was given for the final 4 hours of study. Albumin infusion increased plasma volume by 37%; however, glomerular filtration rate decreased by 32% (p < 0.05). There was no significant change in urine output, sodium excretion, or effective renal plasma flow. Plasma volume expansion with albumin normalized elevated basal levels of aldosterone and plasma renin activity. These findings illustrate that despite substantially increasing plasma volume, colloid infusions reduce glomerular filtration and may limit any associated diuresis. Furthermore, this study demonstrates that hormonal regulation of blood volume remains intact after moderate burn injury.


Rational approach to limiting emergence of antimicrobial drug resistance.
Microbial resistance to the available antimicrobial agents continues to be a major problem with regard to nosocomial and community acquired pathogens. The development of resistance to commonly used antimicrobials is of particular concern when it occurs in pathogenic organisms that cause invasive disease. This has implications on morbidity and mortality of infectious diseases, and will also result in escalated costs of care due to the use of alternative antimicrobials which are often more costly. The increasing frequency of drug resistance has been attributed to combinations of microbial characteristics, selective pressure of antimicrobial use and societal factors that enhance the transmission of drug resistant organisms. The emergence of antibiotic resistant bacteria has generally correlated with the rise and fall of specific antibiotic use in clinical practice. Although the discovery of a new drug temporarily confers therapeutic superiority over bacterial pathogens, the subsequent rapid evolution of resistance limits the duration of the effectiveness of specific agents against pathogens. Surveillance and the development of drug policies that encourage judicious use of antimicrobials will help to minimise the spread of resistant infections. This paper reviews how this dual strategy may be used to control antimicrobial resistance.


[Particulars in the medical treatment of geriatric patients].
During geriatric treatment, the use of medicaments is often important, sometimes harmful, and seldom sufficient by itself. In order for the treatment to be successful, its indication must be well-founded. The dosage has to be adapted, and restraint and therapeutic discipline should always be encouraged. Finally, the anticipated advantages must be greater than the various possible side effects.


How tobacco (Nicotiana tabacum) BY-2 cells cope with Eu(III) - a microspectroscopic study.
The extensive use of lanthanides in science, industry and high-technology products is accompanied by an anthropogenic input of rare earth elements into the environment. Knowledge of a metal's environmental fate is essential for reasonable risk assessment and remediation approaches. In the present study, Eu(III) was representatively used as a luminescent probe to study the chemical environment and to elucidate the molecular interactions of lanthanides with a suspension cell culture of Nicotiana tabacum BY-2. Biochemical methods were combined with luminescence spectroscopy, two-dimensional microspectroscopic mappings, and data deconvolution methods to resolve the bioassociation behavior and spatial distribution of Eu(III) in plant cells. BY-2 cells were found to gradually take up the metal after exposure to 100 μM Eu(III) without significant loss of viability. Time-resolved luminescence measurements were used to specify the occurrence of Eu(III) species as a function of time, revealing the transformation of an initial Eu(III) species into another after 24 h exposure. Chemical microscopy and subsequent iterative factor analysis reveal the presence of four distinct Eu(III) species located at different cellular compartments, e.g., the cell nucleus, nucleolus and cell walls, which could be assigned to intracellular binding motifs. In addition, a special type of bioaccumulation occurs through the formation of a Eu(III)-containing oxalate biomineral, which is already formed within the first 24 hours after metal exposure. Oxalate crystals were also obtained in analogous experiments with Gd and Sm. These results indicate that tobacco BY-2 cells induce the precipitation of metal oxalate biominerals for detoxification of lanthanides, although they also bind to other cellular ligands at the same time.


Postoperative analgesia for shoulder surgery: a critical appraisal and review of current techniques.
Shoulder surgery is well recognised as having the potential to cause severe postoperative pain. The aim of this review is to assess critically the evidence relating to the effectiveness of regional anaesthesia techniques commonly used for postoperative analgesia following shoulder surgery. Subacromial/intra-articular local anaesthetic infiltration appears to perform only marginally better than placebo, and because the technique has been associated with catastrophic chondrolysis, it can no longer be recommended. All single injection nerve blocks are limited by a short effective duration. Suprascapular nerve block reduces postoperative pain and opioid consumption following arthroscopic surgery, but provides inferior analgesia compared with single injection interscalene block. Continuous interscalene block incorporating a basal local anaesthetic infusion and patient controlled boluses is the most effective analgesic technique following both major and minor shoulder surgery. However, interscalene nerve block is an invasive procedure with potentially serious complications and should therefore only be performed by practitioners with appropriate experience.


Investigations of roll-over shape: implications for design, alignment, and evaluation of ankle-foot prostheses and orthoses.
The purpose of this article is to provide an overview of our previous work on roll-over shapes, which are the effective rocker shapes that the lower limb systems conform to during walking. This article is a summary of several recently published articles from the Northwestern University Prosthetics Research Laboratory and Rehabilitation Engineering Research Program on the topic of roll-over shapes. The roll-over shape is a measurement of centre of pressure of the ground reaction force in body-based coordinates. This measurement is interpreted as the effective rocker shape created by lower limb systems during walking. Our studies have shown that roll-over shapes in able-bodied subjects do not change appreciably for conditions of level ground walking, including walking at different speeds, while carrying different amounts of weight, while wearing shoes of different heel heights, or when wearing shoes with different rocker radii. In fact, results suggest that able-bodied humans will actively change their ankle movements to maintain the same roll-over shapes. The consistency of the roll-over shapes to level surface walking conditions has provided insight for design, alignment and evaluation of lower limb prostheses and orthoses. Changes to ankle-foot and knee-ankle-foot roll-over shapes for ramp walking conditions have suggested biomimetic (i.e. mimicking biology) strategies for adaptable ankle-foot prostheses and orthoses.


Poly(anhydride) nanoparticles act as active Th1 adjuvants through Toll-like receptor exploitation.
The mechanisms that underlie the potent Th1-adjuvant capacity of poly(methyl vinyl ether-co-maleic anhydride) nanoparticles (NPs) were investigated. Traditionally, polymer NPs have been considered delivery systems that promote a closer interaction between antigen and antigen-presenting cells (APCs). Our results revealed that poly(anhydride) NPs also act as agonists of various Toll-like receptors (TLRs) (TLR2, -4, and -5), triggering a Th1-profile cytokine release (gamma interferon [IFN-gamma], 478 pg/ml versus 39.6 pg/ml from negative control; interleukin-12 [IL-12], 40 pg/ml versus 7.2 pg/ml from negative control) and, after incubation with dendritic cells, inducing a 2.5- to 3.5-fold increase of CD54 and CD86 costimulatory molecule expression. Furthermore, in vivo studies suggest that NPs actively elicit a CD8(+) T-cell response. Immunization with empty NPs resulted in a significant delay in the mean survival date (from day 7 until day 23 postchallenge) and a protection level of 30% after challenge against a lethal dose of Salmonella enterica serovar Enteritidis. Taken together, our results provide a better understanding of how NPs act as active Th1 adjuvants in immunoprophylaxis and immunotherapy through TLR exploitation.


Plaque inhibition of two commercially available chlorhexidine mouthrinses.
Chlorhexidine (CHX) 0.2% solution is still "the leading oral antiseptic" for controlling gingivitis. Side effects, however, limit the acceptability to users and the long-term employment of a 0.2% CHX antiseptic in preventive dentistry. This stimulated the development of new formulations. The aim of the present study was to assess the effect on plaque inhibition and taste perception of two commercially available mouthrinses (0.12% CHX non-alcohol base with 0.05% cetyl pyridinium chloride (Cpc) versus 0.2% CHX alcohol base). The study was designed as a single-blind, randomized two group parallel experiment, to compare two different commercially available mouthrinses, during a 3-day plaque accumulation model. Forty healthy volunteers were enrolled in the study and received a thorough dental prophylaxis at the beginning of the test period. Over a 72-h experimental non-brushing period, during which subjects abstained from all forms of mechanical oral hygiene, one group (test) used a 15 ml alcohol free 0.12% CHX (=18 mg) mouthrinse on a Cpc base (Perioaid), CHX plus sign in circleCpc), twice daily for 30 s. The other group (control) used a 10 ml 0.2% CHX (=20 mg) mouthrinse on an 11.8% ethanol alcohol base (Corsodyl), CHX plus sign in circleAlc), twice daily for 60 s. After 72 h of plaque formation, the amount of plaque was evaluated. By the use of visual analogue scale, the subjects were asked for their appreciation of the taste of the mouthrinse they had used. The mean plaque index for the CHX plus sign in circleCpc group was 0.97 and for the CHX plus sign in circleAlc group 0.78. After 72 h of non-brushing, there was no significant difference in plaque accumulation between the two groups. The answers to the questions (taste perception and after-taste) showed a statistically significant difference between the two groups. The mean visual analogue scale (VAS) scores for taste appreciation on a scale from very bad to very good taste (0-10) were 5.92 for the CHX plus sign in circleCpc group and 4.10 for the CHX plus sign in circleAlc group (p=0.02). The mean visual analogue scale (VAS) scores for the after-taste on a scale from very short to very long (0-10) were 7.24 for the CHX plus sign in circleCpc group and 5.38 for the CHX plus sign in circleAlc group. Within the limitations of the present study design, it can be concluded that rinsing with a 0.12% CHX mouthrinse on a non-alcohol base with 0.05% Cpc (Perio-Aid) is not significantly different from rinsing with a 0.2% CHX mouthrinse on an alcohol base (Corsodyl). It appears that the subjects appreciated the taste of the non-alcohol CHX solution better but the after-taste of the rinse remained longer in the mouth.


Development of human visual cortex: a balance between excitatory and inhibitory plasticity mechanisms.
Formation of neural circuitry in the developing visual cortex is shaped by experience during the critical period. A number of mechanisms, including N-methyl-D-aspartate (NMDA) receptor activation and gamma-aminobutyric acid (GABA)-mediated inhibition, are crucial in determining onset and closure of the critical period for visual plasticity. Animal models have shown that a threshold level of tonic inhibition must be reached for critical period plasticity to occur and that NMDA receptors contribute to Hebbian synaptic plasticity in the developing visual cortex. There are a number of developmental changes in these glutamatergic and GABAergic mechanisms that have been linked to plasticity; however, those changes have been shown only in animal models, and their development in the human visual cortex is not known. We have addressed this question by studying the expression of the major glutamatergic receptors, GABA(A) receptors, and glutamic acid decarboxylase (GAD) isoforms during the first 6 years of postnatal development of human visual cortex. There are significant changes in the expression of these proteins during postnatal development of human visual cortex. The time course of the changes is quite prolonged and suggests that it may set the pace for the prolonged critical period in human visual development. The changes also affect the nature of spatial and temporal integration in visual cortical neurons and thereby contribute to the maturation of visual functions.