[A Case of Ruptured Internal Carotid-Posterior Communicating Artery Aneurysm Associated with Acute Subdural Hematoma, Extending from the Interhemispheric Space to the Posterior Fossa].
A 69-year-old woman was admitted to our hospital because of a sudden severe headache without a history of head trauma. CT and MRI revealed an acute subdural hematoma (ASDH) extending from the right interhemispheric space to the posterior fossa bilaterally, with a small amount of subarachnoid hemorrhage that was predominantly localized to the left side of the basal cistern. CT angiogram demonstrated a long protruding ruptured aneurysm at the junction of the right internal carotid and posterior communicating arteries (IC/PC AN) with a posteroinferior projection, associated with a small bleb located near the tentorial edge close to the ipsilateral posterior clinoid process, for which she received clipping surgery. Though rare, IC/PC AN could cause pure or nearly pure ASDH in the above-mentioned distribution. Therefore, in patients with such ASDH, especially without a history of head injury or precise information regarding the situation at the time of onset, urgent imaging evaluation and early intervention are essential to prevent devastating re-rupture events.


Plant steroid hormones produced under Ni stress are involved in the regulation of metal uptake and oxidative stress in Brassica juncea L.
Brassinosteroids (BRs) are involved in the amelioration of various biotic and abiotic stresses. With an aim to explore the role of BRs under heavy metal stress, plants of Brassica juncea L. were grown in pots. The plants were subjected to various concentrations of Nickel metal (0.0, 0.2, 0.4 and 0.6 mM) and harvested on 60th day in order to observe the expression of these hormones. The isolated BRs from the leaves of Brassica plants characterized by GC-MS include 24-Epibrassinolide (24-EBL), Castasterone, Dolicholide and Typhasterole. The effect of isolated 24-EBL was studied on Ni metal uptake and antioxidative defense system in 60 d old plants of Brassica. It was observed that 24-EBL significantly increased the activities of stress ameliorating enzymes and lowered the metal uptake in plants. This is the first report in B. juncea L. plants showing the expression of BRs under metal treatments and effect of the isolated 24-EBL on metal uptake and in oxidative stress management.


Systems of reinforcement and drug dependence.
Experimental data are presented in support of the hypothesis on the role of positive and negative reinforcing systems in the mechanism of drug dependence. Drugs with abuse potential (DAP) may change the manner of response to negative emotional stimuli, activate positive emotional reactions in animals, and possess primary reinforcing properties. Reward and punishment systems respond sensitively to withdrawal from DAP or antagonist administration. Catecholaminergic and peptidergic processes are of importance in the mechanisms of the emotionally positive action of DAP.


Pelvic floor muscle training in mother fit healthcare centres: a best practice implementation project.
Pelvic floor dysfunctions are common complications during the childbirth process. Pelvic floor muscle training (PFMT) has been used as a conservative treatment in both prenatal and postnatal women. The aim of this project was to assess the compliance with evidence-based criteria regarding PFMT among prenatal and postnatal women in mother fit healthcare centres. The current study was performed in three phases. Phase 1: stakeholder engagement and team establishment and baseline audit. Phase 2: Design and implementation of strategies to improve PFMT among prenatal and postnatal women in mother fit healthcare centres. Phase 3: Follow-up audit. Baseline and follow-up audit with six yes/no audit questions was performed using the Joanna Briggs Institute Practical Application of Clinical Evidence System. Fifteen midwives involved in prenatal and postnatal healthcare centres and 60 pregnant women participated in this study. The compliance for each audit criterion found in the baseline audit was low (0-7%). However, after implementation of the project and training the trainers (midwives), the compliance increased significantly (77-100%). The current study demonstrated that we have some problems in the healthcare system to teach PFMT to pregnant women. We should also implement similar projects in all healthcare centres country wide. Healthcare system managers should be aware of the value of holding PFMT courses for pregnant women and it is recommended to use pelvic floor physiotherapists to train midwives.


A case of Candida parapsilosis endocarditis.
The authors report a case of Candida parapsilosis endocarditis in a non-drug-addicted young patient who, 6 months before the manifestation of endocarditis, was submitted to cardiac surgery. This is the first report from Greece and the second found in the accessible literature.


Interleukin-2 inhibits sodium currents in human muscle cells.
The effect of the T cell growth factor interleukin-2 (IL-2) on muscular Na+ channels was studied in myoballs produced from primary human muscle cultures. The transient Na+ inward currents of the myoballs, elicited by repetitive stimulation at 1 Hz and recorded in the whole-cell mode, were inhibited by IL-2 applied to the external solution, the half maximum effect occurring at 300 U/ml. The effect was complete within 5 s and was totally reversible, the on and off effects having identical time courses. The h infinity curve was shifted in negative direction indicating that the mechanism of IL-2 action is a conversion of the Na+ channels into a state of inactivation. The reaction of the IL-2 solution with an anti IL-2 antibody neutralized the inhibitory effect on the Na+ currents, indicating a specific effect of the peptide growth factor interleukin-2 on muscular Na+ channels. The connection of IL-2 and Na+ channels may be important in inflammatory processes of muscle and nerve.


KLF5 inhibition overcomes oxaliplatin resistance in patient-derived colorectal cancer organoids by restoring apoptotic response.
Oxaliplatin resistance is a major challenge in the treatment of colorectal cancer (CRC). Many molecular targeted drugs for refractory CRC have been developed to solve CRC drug resistance, but their effectiveness and roles in the progression of CRC and oxaliplatin resistance remain unclear. Here, we successfully constructed CRC PDOs and selected the Kruppel-like factor 5 (KLF5) inhibitor ML264 as the research object based on the results of the in vitro drug screening assay. ML264 significantly restored oxaliplatin sensitivity in CRC PDOs by restoring the apoptotic response, and this effect was achieved by inhibiting the KLF5/Bcl-2/caspase3 signaling pathway. Chromatin immunoprecipitation (ChIP) and luciferase reporter assays verified that KLF5 promoted the transcription of Bcl-2 in CRC cells. KLF5 inhibition also overcame oxaliplatin resistance in xenograft tumors. Taken together, our study demonstrated that ML264 can restore oxaliplatin sensitivity in CRC PDOs by restoring the apoptotic response. KLF5 may be a potential therapeutic target for oxaliplatin-resistant CRC. PDOs have a strong potential for evaluating inhibitors and drug combination therapy in a preclinical environment.


A flow cytometric method for the measurement of phagocytosis by polymorphonuclear leucocytes.
A new method for the measurement of phagocytosis of Candida albicans by human polymorphonuclear leucocytes (PMN) is described using a fluorescence activated cell sorter. We have used acridine orange to discriminate between PMN which have internalised yeast particles and those which have not. This method allows accurate measurement of particle phagocytosis as an event distinct from particle adherence. It also permits detailed examination of the kinetics of phagocytosis, the study of which is likely to be of value in the investigation of diseases where abnormalities of PMN function are suspected.


Caffeine antinociception in the rat hot-plate and formalin tests and locomotor stimulation: involvement of noradrenergic mechanisms.
The present study examined antinociception produced by systemic administration of caffeine in the rat hot-plate (HP) and formalin tests and addressed several aspects of the mechanism of action of caffeine. Locomotor activity was monitored throughout. Caffeine produced a dose-related antinociception the HP (50-100 mg/kg) and formalin tests (12.5-75 mg/kg). When observed during the formalin test, caffeine stimulated locomotor activity between 12.5 and 50 mg/kg; this was followed by a depression in activity at 75 mg/kg. Caffeine did not produce an anti-inflammatory effect as determined by hindpaw plethysmometry, suggesting that antinociception was not secondary to an anti-inflammatory action. Peripheral co-administration of caffeine with the formalin did not produce antinociception, suggesting a predominant central rather than peripheral site of action for caffeine. Naloxone (10 mg/kg) did not reduce the antinociceptive or locomotor stimulant effects of caffeine, suggesting a lack of involvement of endogenous opioids in these actions. Phentolamine (5 mg/kg) enhanced antinociception by caffeine in both the HP and formalin tests, but inhibited locomotor stimulation. Prazosin (0.15 mg/kg) mimicked the action of phentolamine on locomotor stimulation, but idazoxan (0.5 mg/kg) mimicked the action of phentolamine on antinociception in the formalin test. These observations suggest an involvement of different alpha-adrenergic receptors in the two actions of phentolamine. Microinjection of 6-hydroxydopamine (6-OHDA) into the locus coeruleus, which depleted noradrenaline (NA) in the spinal cord and forebrain, inhibited the action of caffeine in the HP test. This was mimicked by intrathecal 6-OHDA which depleted NA in the spinal cord, but not by microinjection of 6-OHDA into the dorsal bundle which depleted NA in the forebrain. These results suggest an integral involvement of noradrenergic mechanisms in the antinociceptive action of caffeine in the HP and formalin tests and in locomotor stimulation, but the nature of this involvement differs for the 3 end points.


The synthesis of hyaluronic acid by human synovial fibroblasts is influenced by the nature of the hyaluronate in the extracellular environment.
Various cell lines of human synovial fibroblasts derived from synovium obtained at the time of biopsy or total joint-replacement surgery have been established. The synthesis of 3H-labelled hyaluronic acid (HA) in these cells has been determined, and the effects of adding HA of varying molecular size to the cultured cells examined. The results obtained clearly show that the in vitro synthesis of HA by these cells is influenced by the concentration and molecular weight (MW) of the HA in their extracellular environment. Synovial fibroblasts derived from an osteoarthritic joint demonstrated the most marked response on exposure to exogenous HA, showing a stimulation of HA synthesis with preparations of weight-average molecular weight (Mw) greater than 5 X 10(5) in a concentration dependent manner. HA preparations with Mw less than 5 X 10(5) showed little or no effect except at high concentrations where a suppression of biosynthesis was observed. A model to explain these findings is proposed.


Enhanced articulatory speed in ambidexters.
The possibility that incomplete speech lateralization might be related to reduced articulatory speed was investigated in two studies with normal, young adults. Contrary to expectations, speech was produced faster by ambidextrous subjects than by either strongly left- or strongly right-handed subjects under a number of conditions. However, there were no differences in speech lateralization between these groups when lateralization was measured using dichotic listening scores. In a second study, the superiority of ambidextrous subjects was found to extend to certain manual movement tasks as well.


Ionically crosslinked chitosan/poly(acrylic acid) hydrogels with high strength, toughness and antifreezing capability.
A dual physically crosslinking (DPC) strategy is used to construct hydrogels with ultrahigh strength. First, polyelectrolyte complex (PEC) hydrogels were prepared through in situ polymerization of acrylic acid monomers in chitosan solutions. Subsequently, cations and anions were introduced into the PEC hydrogels to form strong electrostatic interactions with the polymer chains. The mechanical properties of the DPC hydrogels strongly depended on the ionic concentration and the valence state of the loading ions. The tensile strength of DPC-Na 8-20-2.0, DPC-Mg 8-20-2.0, DPCAl 8-20-2.0 and DPC-Sul 8-20-0.8 reached to 2.36, 12.59, 65.1 and 2.80 MPa, respectively, which were significantly higher than that of PEC 8-20 (0.29 MPa). Moreover, DPC-Na, DPC-Mg and DPC-Sul still maintained a good flexibility. Specifically, hydrogels of DPC-Ca exhibited ionic conductivity and freeze tolerance, which could be cooled to -20 °C without freezing. The DPC strategy opens an avenue to fabricate hydrogels with outstanding mechanical properties.


The changes of endotracheal tube intracuff pressures after ear and head and neck surgery-related positions: a prospective observational study.
The cuff of an endotracheal tube seals the airway to facilitate positive-pressure ventilation and reduce subglottic secretion aspiration. However, an increase or decrease in endotracheal tube intracuff pressure can lead to many morbidities. The main purpose of this study is to investigate the effect of different head and neck positions on endotracheal tube intracuff pressure during ear and head and neck surgeries. A total of 90 patients undergoing elective right ear (Group 1: n=30), left ear (Group 2: n=30) or head and neck (Group 3: n=30) surgery were involved in the study. A standardized general anesthetic was given and cuffed endotracheal tubes by the assistance of video laryngoscope were placed in all patients. The pilot balloon of each endotracheal tube was connected to the pressure transducer and standard invasive pressure monitoring was set to measure intracuff pressure values continuously. The first intracuff pressure value was adjusted to 18.4mmHg (25cm H2O) at supine and neutral neck position. The patients then were given appropriate head and neck positions before related-surgery started. These positions were left rotation, right rotation and extension by under-shoulder pillow with left/right rotation for Groups 1, 2 and 3, respectively. The intracuff pressures were measured and noted after each position, at 15th, 30th, 60th, 90th minutes and before the extubation. If intracuff pressure deviated from the targeted value of 20-30cm H2O at anytime, it was set to 25cm H2O again. The intracuff pressure values were increased from 25 to 26.73 (25-28.61) cm H2O after left neck rotation (p=0.009) and from 25 to 27.20 (25.52-28.67) cm H2O after right neck rotation (p=0.012) in Groups 1 and 2, respectively. In Group 3, intracuff pressure values at the neutral position, after extension by under-shoulder pillow and left or right rotation were 25, 29.41 (27.02-36.94) and 34.55 (28.43-37.31) cm H2O, respectively. There were significant differences between the neutral position and extension by under-shoulder pillow (p<0.001), and also between neutral position and rotation after extension (p<0.001). However, there was no statistically significant increase of intracuff pressure between extension by under-shoulder pillow and neck rotation after extension positions (p=0.033). Accessing the continuous intracuff pressure value measurements before and during ear and head and neck surgeries is beneficial to avoid possible adverse effects/complications of surgical position-related pressure changes.


"It's not easy to change the mentality": Challenges to sex education delivery for persons with intellectual disability.
Sex education for people with intellectual disability is in its infancy in countries where their sexual expression is still considered a taboo. This study aimed at exploring the experiences of professionals involved in providing sex education to people with intellectual disability across a number of services in a Maltese context. Qualitative research was conducted using interviews to explore professionals' experiences of sex education provision. Amid a persistent culture of sexual suppression, desexualisation, infantalization and control remain barriers to sex education delivery. Findings underline the need for services to have clear policies that aid professionals in delivering sex education, while mitigating conflicts encountered. Recommendations highlight sex positive environments and services, with a pro-active approach to sex education. In spite of internationally binding legislation and local policies, further changes need to be made on the ground to provide persons with intellectual disability with effective access to sex education.


Location of the TEMPO moiety of TEMPO-PC in phosphatidylcholine bilayers is membrane phase dependent.
The (2,2,6,6-tetramethylpiperidin-1-yl)oxyl (TEMPO) moiety tethered to the headgroup of phosphatidylcholine (PC) lipid is employed in spin labeling electron paramagnetic resonance spectroscopy to probe the water dynamics near lipid bilayer interfaces. Due to its amphiphilic character, however, TEMPO spin label could partition between aqueous and lipid phases, and may even be stabilized in the lipid phase. Accurate assessment of the TEMPO-PC configuration in bilayer membranes is essential for correctly interpreting the data from measurements. Here, we carry out all-atom molecular dynamics (MD) simulations of TEMPO-PC probe in single-component lipid bilayers at varying temperatures, using two standard MD force fields. We find that, for a dipalmitoylphosphatidylcholine (DPPC) membrane whose gel-to-fluid lipid phase transition occurs at 314 K, while the TEMPO spin label is stabilized above the bilayer interface in the gel phase, there is a preferential location of TEMPO below the membrane interface in the fluid phase. For bilayers made of unsaturated lipids, 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), which adopt the fluid phase at ambient temperature, TEMPO is unequivocally stabilized inside the bilayers. Our finding of membrane phase-dependent positioning of the TEMPO moiety highlights the importance of assessing the packing order and fluidity of lipids under a given measurement condition.


Can I control my bowel symptoms myself? The experience of controlling defaecation dysfunction among patients with rectal cancer after sphincter-saving surgery: a qualitative study.
To explore the experience of controlling defaecation dysfunction among patients with rectal cancer after sphincter-saving surgery. This study applied a descriptive qualitative design. Thirty-six patients with rectal cancer were given semi-structured interviews in mainland China from February to July in 2019 after sphincter-saving surgery. Participants were recruited by purposive sampling. The thematic analysis approach was applied to analyse the transcripts. Three major themes emerged from the data were "having motivations of controlling defecation dysfunction", "using strategies of controlling defecation dysfunction" and "facing barriers of controlling defecation dysfunction". Defaecation dysfunction makes obvious problems for patients after sphincter-saving surgery, although patients tried some self-care methods to cope with the defaecation dysfunction, some barriers still exist in the process of self-controlling of bowel symptoms. There is a strong demand for a systematic and scientific guideline for the self-management of defaecation dysfunction.


The significance of perineural invasion in early-stage cervical cancer.
Cervical cancer spreads directly and through lymphatic and vascular channels. Perineural invasion is an alternative method of spread. Several risk factors portend poor prognosis and inform management decisions regarding adjuvant therapy. To evaluate the incidence and significance of PNI in early cervical cancer. Retrospective chart review of early-stage cervical cancer patients (IA-IIA) from 1994 to 2009. One hundred ninety two patients were included, 24 with perineural invasion in the cervical stroma (cases) and 168 without (controls). The mean age of the cases was 53 years, versus 45.9 in the controls (P=0.01). PNI was associated with more adjuvant therapy (P=0.0001), a higher stage (P=0.005), a larger tumor size (≥ 4 cm) (P<0.0001), lymphovascular space invasion (P=0.002), parametrial invasion (P<0.0001) and more tumor extension to the uterus (P=0.015). On multivariate analysis using an adjusted hazard ratio, risk factors for recurrence included grade (HR, 95% CI; 3.61, 1.38-9.41) and histopathology (HR, 95% CI; 2.85, 100-8.09). Similarly, risk factors for death included grade (HR, 95% CI; 3.43, 1.24-9.49) and histopathology (HR, 95% CI; 3.71, 1.03-13.33). Perineural invasion was not identified as an independent risk factor for either recurrence or death. The mean follow up time was 56 months. There was no significant difference in recurrence (P=0.601) or over-all survival (P=0.529) between cases and controls. While perineural invasion was found to be associated with multiple high-risk factors, it was not found to be associated with a worse prognosis in early cervical cancer.


Nitric oxide inhibits resting sphincter of Oddi activity.
The sphincter of Oddi has basal myogenic phasic activity that is modulated by neural and hormonal pathways. Stimulatory innervation to this organ is cholinergic, whereas the inhibitory pathways are unknown. Nitric oxide (NO), generated from L-arginine, relaxes gastrointestinal smooth muscle in vitro. We, therefore, hypothesized that resting sphincter of Oddi and duodenal motilities are regulated by a NO-mediated inhibitory pathway. In 23 anesthetized prairie dogs, systemic blood pressure and sphincter of Oddi and duodenal motilities were monitored during systemic infusion of N omega-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide synthase. L-NAME was infused alone and simultaneously with excess D- and L-arginine. L-NAME alone and L-NAME with D-arginine produced hypertension and increased sphincter of Oddi and duodenal motilities. L-arginine blocked these increases, suggesting that baseline sphincter of Oddi and duodenal motility regulation involves the generation of NO from L-arginine. We conclude that baseline sphincter of Oddi phasic activity is regulated by cholinergic stimulatory and NO-mediated inhibitory neural pathways.


Assisting in Microsurgery: Operative and Technical Considerations.
Microsurgery is technically challenging, typically requiring a primary surgeon and an assistant to complete several key operative steps. These may include manipulation of fine structures, such as nerves or vessels in preparation for anastomosis; stabilization of the structures; and needle driving. Even seemingly mundane tasks of suture cutting and knot tying require fine coordination between the primary surgeon and assistant in the microsurgical environment. Although prior literature discusses the implementation of microsurgical training centers at academic institutions and residency programs, there is a paucity of work describing the role of the assistant surgeon in a microsurgery operation. In this surgical technique article, the authors discuss the role of the assisting surgeon in microsurgery, with recommendations for trainees and attendings alike.


Afatinib in Non-Small Cell Lung Cancer Harboring Uncommon EGFR Mutations Pretreated With Reversible EGFR Inhibitors.
Afatinib, an irreversible ErbB family blocker, is approved for treatment of patients with previously untreated non-small cell lung cancer (NSCLC) harboring activating epidermal growth factor receptor (EGFR) mutations. Efficacy of afatinib in EGFR tyrosine kinase inhibitor-naïve (TKI-naïve) patients with uncommon EGFR mutations (other than exon 19 deletions or exon 21 point mutations) has been reported; however, efficacy in TKI-pretreated patients with uncommon EGFR mutations is unknown. In the afatinib compassionate use program (CUP), patients with advanced or metastatic, histologically confirmed NSCLC progressing after at least one line of chemotherapy and one line of EGFR-TKI treatment were enrolled. Demographic data, mutation type, response rates, time to treatment failure (TTF), and safety in patients harboring uncommon EGFR mutations were reported. In 60 patients (63% female, median age 63 years [range: 30-84 years]), a total of 66 uncommon EGFR mutations including 30 T790M mutations were reported (18.4% and 11%, respectively, of known EGFR mutations within the CUP). Most patients (67%) received afatinib as third- or fourth-line treatment. Median TTF was 3.8 months (range: 0.2 to >24.6 months; p = .244) in patients with uncommon mutations compared with 5.1 months (range: 0.1 to >21.1 months) in patients with common mutations (n = 165). Pronounced activity was observed with E709X mutations (TTF >12 months). No new safety signals were detected. Afatinib is clinically active and well tolerated in many TKI-pretreated NSCLC patients harboring uncommon EGFR mutations. Compared with results reported in TKI-naïve patients, activity was also indicated in patients with T790M and exon 20 insertion mutations.


Monitoring of periodic anaerobic digestion with flow-through array of miniaturized ion-selective electrodes.
In the last few years Electronic tongues (ETs) based on various sensor arrays were applied to the monitoring of various fermentation processes, as devices capable of fast, inexpensive, automated and on-line control. In this work a novel application of ET is proposed--flow-through array of miniaturized ion-selective electrodes (ISEs) was used for the analysis of samples obtained during anaerobic digestion (methane fermentation) performed in periodic conditions. The samples were classified according to their Chemical Oxygen Demand (COD) and Volatile Fatty Acid (VFA) content. Moreover, for this application a novel design of ISEs was developed, which is fully compatible with flow-through modules for sensor measurements.


Cycling of organic and inorganic sulphur in a chestnut oak forest.
Sulfur (S) cycling in a chestnut oak forest on Walker Branch Watershed, Tennessee, was dominated by geochemical processes involving sulfate. Even though available SO 42- was present far in excess of forest nutritional requirements, the ecosystem as a whole accumulated ∼60% of incoming SO4-S. Most (90%) of this accumulation occurred by SO 42- adsorption in sesquioxide-rich subsurface soils, with a relatively minor amount accumulating and cycling as SO 42- within vegetative components. Organic sulfates are thought to constitute a large proportion of total S in surface soils, also, and to provide a pool of readily mineralized available S within the ecosystem.


Synthesis and evaluation of 3- and 7-substituted geranylgeranyl pyrophosphate analogs.
Protein prenyl transferases have been a focus of anti-cancer drug discovery in recent years due to their roles in post-translational modification of small GTP binding proteins. Attention is now turning to the development of GGTase I inhibitors. Here, we present the synthesis and biological evaluation of four GGPP analogs versus mammalian GGTase I and the discovery that 7-allyl GGPP is a surprisingly efficient GGTase I substrate.


High-throughput method for a kinetics analysis of the high-pressure inactivation of microorganisms using microplates.
Using microplates as pressure and cultivation vessels, a high-throughput method was developed for analyzing the high-pressure inactivation kinetics of microorganisms. The loss of viability from a high-pressure treatment, measured based on the growth delay during microplate cultivation, showed reproducibility with the conventional agar plate method and was applicable for the kinetics analysis.


Determination of antibiotic residues in frozen poultry by a solid-phase dispersion method using liquid chromatography-triple quadrupole mass spectrometry.
Importation of poultry produce into Nigeria through its land borders has heightened, notwithstanding the government's ban on such products. This study examined imported frozen poultry products for antibiotic residues considering their health implications. A solid-phase extraction method using liquid chromatography-tandem mass spectrometry in the positive and negative electrospray ionisation and the multiple reaction monitoring modes were employed. The antibiotics were extracted with acetonitrile-dichloromethane. Chromatographic separation was on Waters Acquity UPLC® BEH C18 column with acetonitrile, and water gradient and the antibiotics analysed using Electrospray positive ionisation polarity switch in a single run of fourteen minutes. Residues of nineteen (19) antibiotics were found in the three different matrices at different levels with varying detection frequencies ranging between 2 and 4% (sulfamoxole, penicillin-G, albendazole and phebendazole) and 14-54% for all the other antibiotics. The highest number of violative samples was found in the turkey gizzard and chicken muscle. Sulfixosazole had the highest percentage violation of 80.00% in turkey gizzard while sulfamethoxazole, notwithstanding its lower frequency in chicken muscle had highest maximum concentration and 100% violation. The presence of these drugs, however, does not pose any immediate health risk.


Preclinical toxicity of innovative molecules: In vitro, in vivo and metabolism prediction.
The lack of predictivity of animal's models has increased the failure rate of drug candidates. Thus, the reversion of this scenario using preliminary in vitro assays and metabolism prediction can reduce the unnecessary use of animals, as well as predict toxic effects at preclinical and clinical stages. The present study aimed to evaluate safety of four biologically active molecules (RN104, RI78, ICH, PCH) with potential therapeutic applications synthesized in our laboratory. Initially, we used MTT cytotoxicity against A549, H9C2, HepG2, LLC-PK1 and NEURO-2 cell lines. RN104 showed the lowest cytotoxicity and further studies were conducted with it. The neutral red (NR) test was performed according to OECD-129 and then acute toxicity test (OECD-423). According to NR results we administered at 300 mg/kg on animals; however, no toxic effect was observed, while 2,000 mg/kg resulted in the death of one animal per group. After, metabolism prediction studies, performed using both ligand-based and structure-based, suggests three potential metabolites. In silico results suggested that potential metabolites could be fast eliminated and, then, this could be an explanation for lower observed toxicity in in vivo experiments. The results showed limitations of the NR as a predictor of the initial dose for the acute toxicity study, which may be related to metabolism. Therefore, the combination of theoretical and experimental studies is relevant to a general understanding of new molecule's toxicity.


Multicentric reticulohistiocytosis.
Multicentric reticulohistiocytosis (MR) is a rare histiocytic systemic disease mainly affecting the skin and synovia. To present the main and newest aspects of MR. A review of the current medical literature, composing a retrospective study of 96 case reports published world-wide since 1977. Facial and hand skin nodules with symmetric arthritis were the most frequent clinical presentation. The pathognomonic sign of'coral beads' and vermicular erythematous lesions bordering nostrils, seems to be very characteristic of MR. There is a significant association with cancer. Histiocytic cells of MR are best characterized on immunohistochemistry by its immunoreactivity for vimentin, CD68 and CD45 and non-reactivity for S-100 protein, CD34 and factor XIIIa. Although the outcome of the disease is usually very limiting for the patients, treatment with immunosuppressive drugs may be useful. MR is a unique histiocytic proliferative disease with a peculiar immunophenotypic pattern and distinctive clinical and histopathological aspects.


Does a balanced transfusion ratio of plasma to packed red blood cells improve outcomes in both trauma and surgical patients? A meta-analysis of randomized controlled trials and observational studies.
The effect of high transfusion ratios of fresh frozen plasma (FFP): packed red blood cell (RBC) on mortality is still controversial. Observational evidence contradicts a recent randomized controlled trial regarding mortality benefit. This is an updated meta-analysis, including a non-trauma cohort. Patients were grouped into high vs. low based on FFP:RBC ratio. Primary outcomes were 24-h and 30-day/in-hospital mortality. Secondary outcomes were acute respiratory distress syndrome and acute lung injury rates. Random model and leave-one-out-analyses were used. In 36 studies, lower ratio showed poorer 24-h and 30-day survival (p < 0.001). In trauma and non-trauma settings, a lower ratio was associated with worse 24-h and 30-day mortality (P < 0.001). A ratio of 1:1.5 provided the largest 24-h and 30-day survival benefit (p < 0.001). The ratio was not associated with ARDS or ALI. High FFP:RBC ratio confers survival benefits in trauma and non-trauma settings, with the highest survival benefit at 1:1.5.


Early hepatic stellate cell activation predicts severe hepatitis C recurrence after liver transplantation.
Only a subset of hepatitis C virus (HCV)-infected patients develop progressive hepatic fibrosis after liver transplantation (LT). Hepatic stellate cell (HSC) activation is a pivotal step in hepatic fibrosis and precedes clinically apparent fibrosis. We determined whether early HSC activation, measured in 4-month protocol post-LT biopsies, is predictive of subsequent development of more histologically severe recurrence of HCV. Early (4 month) post-LT HSC activation, as measured by alpha-smooth muscle actin (alpha-SMA) staining, was determined in liver biopsies from recipients with severe (fibrosis score > or = 2, n = 13) and with mild (fibrosis score of 0, n = 13) recurrence of HCV at one-year post-LT. Immunohistochemical staining for alpha-smooth muscle actin (alpha-SMA) was used to generate HSC activation scores (regional and total). Total HSC activation scores at 4 months were similar in patients with severe and mild HCV recurrence (3.9 +/- 2.0 vs. 2.7 +/- 2.2, P = 0.2). Regional HSC activation, assessed as parenchymal (zones 1, 2, and 3) or mesenchymal (portal tracts and fibrous septa), was different between the study groups, with higher mesenchymal scores predictive of progression. No patients in the mild recurrence group had detectable mesenchymal alpha-SMA staining vs. 46% (6/13) of patients with severe recurrence (P < 0.01). Mesenchymal activation of HSC had a specificity and positive predictive value of 100% for development of progressive fibrosis in liver allografts of patients with hepatitis C. In conclusion, early activation of mesenchymal HSCs is a marker for progressive fibrosis in patients with hepatitis C post-LT and may help select patients who would benefit from HCV or HSC-targeted therapy.


Molecular cloning of pig Rad51, Rad52, and Rad54 genes, which are involved in homologous recombination machinery.
The low rate of homologous recombination in somatic cells is considered to be an urgent issue. Therefore, we molecularly cloned three genes that relate to efficient homologous recombination. Polymerase chain reaction (PCR) was performed to isolate candidate cDNA fragments from a pig spleen cDNA library with the corresponding primer sets deduced from multiple alignment analysis of other mammalian genes. A 5'- and 3'-RACE PCR experiment was performed to determine the complete cDNA sequences. The complete cDNA sequences of the pig RAD51, RAD52, and RAD54 genes, which are closely related to homologous recombination events, were identified using molecular cloning technique. The cDNA sequences of three genes were successfully isolated by PCR-based methods. As a result, we determined the sequences of pig RAD51 (1663 bp, 339 aa), RAD52 (1884 bp, 406 aa), and RAD54 (2884 bp, 747 aa). The nucleic acid sequence homologies of the pig RAD51, RAD52, and RAD54 genes compared with the corresponding human genes were 92.9%, 77.3%, and 90.0%; the corresponding amino acid sequence homologies were 98.8%, 71.1%, and 95.0%, respectively. The knockout of alpha-1,3-galactosyltransferase in pigs resulted in a drastic reduction in xenoantigenicity. However, other xenoantigens, in particular, the non-Gal antigens, also need to be down-regulated. Gene transfer to alter expression levels of these recombination-related molecules and/or ex ante evaluation of expression profiles of these genes in primary cultures of somatic cells constitute a new approach to enhancing homologous recombination events during the production of gene knockout pigs.


Angiographic evaluation of conjoined twins.
Two sets of conjoined twins were studied by angiocardiography and cerebral angiography. Conjoined heart was demonstrated in the thoracopagus twins and surgical separation was impossible. Cerebral angiography disclosed the separate circulations in craniopagus twins and surgical separation was performed.


Febrile Neutropenia in Patients with Solid Tumors Undergoing Intravenous Chemotherapy.
Febrile neutropenia (FN) is a potentially life-threatening complication of systemic chemotherapy (CT) that often requires hospital admission. Delay in diagnosis and treatment are associated with higher morbidity and mortality. We aimed to determine the factors that influence FN episodes outcomes in the emergency room (ER). This was a retrospective study of all FN episodes (with a collected blood culture [BC]) that occurred between 2012 and 2016 at our institution. FN was defined as a temperature ≥38°C and an absolute neutrophil count (ANC) <1,000/μL, expected to decrease to <500/μL in the following week. Between 2012 and 2016, there were 173 FN episodes in 153/1,947 patients treated with intravenous CT. Most of these episodes (n = 121, 70%) were diagnosed in the ER, 29 in the outpatient clinic, and 23 as inpatients. In the ER, the median time was 36 min from hospital nurse triage to medical observation, and 52 min from medical observation to complete blood count specimen collection. There was a positive BC in 33 FN episodes, 72% with Gram-negative bacteria. A total of 160 FN episodes led to hospital admission and 13 were treated as outpatients. Mortality associated with the FN episode was 15% and an ANC <100/μL was predictive of increased mortality. This study confirms that FN is a serious and common complication of IV CT which must be diagnosed and treated promptly. Profound neutropenia was the only predictive factor of mortality.


An innovative approach in the management of alveolar clefts with bone graft harvest from maxillary tuberosity and mandibular wisdom molar odontectomy sites: A case report.
Alveolar bone grafting is a complex procedure utilized in alveolar cleft repair; however, the ideal site of bone graft material remains highly debated. In this study, we describe the management of a 14-year-old girl with bilateral alveolar clefts using alternative intraoral donor sites for bone graft harvest.


Association of Pregnancy Characteristics and Maternal Mortality With Amniotic Fluid Embolism.
Amniotic fluid embolism (AFE) is an uncommon pregnancy complication but is associated with high maternal mortality. Because of the rarity of AFE, associated risks factors and maternal outcomes have been relatively understudied. To examine the clinical, pregnancy, and delivery characteristics and the maternal outcomes related to AFE in a recent period in the US. This retrospective cohort study examined hospital deliveries from January 1, 2016, to December 31, 2019, from the Healthcare Cost and Utilization Project's National Inpatient Sample. The primary outcome was clinical, pregnancy, and delivery characteristics of AFE, assessed with a multivariable binary logistic regression model. The coprimary outcome was failure to rescue, defined as maternal mortality after AFE. Associations with other severe maternal morbidity indicators and failure to rescue per clinical and pregnancy characteristics were also assessed. A total of 14 684 135 deliveries were examined, with AFE diagnosed in 880 women, corresponding to an incidence rate of 6.0 per 100 000 deliveries. The cohort-level median patient age was 29 years (IQR, 25-33 years). In a multivariable analysis, (1) patient factors of older age, Asian and Black race, Western US region, pregestational hypertension, asthma, illicit substance use, and grand multiparity; (2) pregnancy factors of placental accreta spectrum (PAS), placental abruption, uterine rupture, polyhydramnios, chorioamnionitis, preeclampsia, fetal growth restriction, and fetal demise; and (3) delivery factors of early gestational age, cervical ripening, cesarean delivery, operative delivery, and manual removal were associated with AFE. Among these characteristics, PAS had the largest association with AFE (adjusted odds ratio [aOR], 10.01; 95% CI, 7.03-14.24). When stratified by the PAS subtypes, more severe forms of PAS had a greater association with AFE (aOR for increta and percreta, 17.35; 95% CI, 10.21-28.48; and aOR for accreta, 7.62; 95% CI, 4.83-12.01). Patients who had AFE were more likely to have coagulopathy (aOR, 24.68; 95% CI, 19.38-31.44), cardiac arrest (aOR, 24.56; 95% CI, 17.84-33.81), and adult respiratory distress syndrome (aOR, 10.72; 95% CI, 8.09-14.20). The failure-to-rescue rate after AFE was 17.0% overall. However, the failure-to-rescue rate exceeded 30% when AFE co-occurred with other severe maternal morbidity indicators: 45.8% for AFE, cardiac arrest, and coagulopathy; 43.2% for AFE, shock, and cardiac rhythm conversion; and 38.6% for AFE, cardiac arrest, coagulopathy, and shock. The failure-to-rescue rate after AFE also exceeded 30% when AFE occurred in the setting of placental pathology: 42.9% for AFE and PAS and 31.3% for AFE and placental abruption. This contemporaneous, national-level analysis validated previously known risk factors for AFE and confirmed the dismal outcomes of pregnancy complicated by AFE. The association between PAS and AFE, which was not previously reported, warrants further investigation.


Enhancing copper binding property of compost-derived humic substances by biochar amendment: Further insight from two-dimensional correlation spectroscopy.
Little is known about the environmental impacts of biochar (BC) amendment on the immobilization of heavy metals in compost-treated fields. In consequence, this study was designed to explore the effects of BC amendment on the copper (Cu) binding properties of compost-derived humic substances (HS). To this end, unamended and 4.7% (by wet weight) BC-amended compost were incubated in parallel with regular wetting for 6 months. The stability constants for Cu binding, calculated based on the fluorescence quenching technique, were higher for the compost-derived HS with versus without BC amendment. The result suggests that BC addition to compost may intensify the immobilization of heavy metals in a compost-treated field. Copper binding efficacy increased after the incubation with or without BC amendment. However, the enhanced Cu binding efficacy of the BC-amended HS was preserved even after the long-term incubation. Two-dimensional correlation spectroscopy revealed that the short wavelength fulvic-like fluorescence followed by humic-like fluorescence were preferentially associated with Cu binding. However, the range of wavelengths that tracked the binding of Cu were modified after BC amendment. This study evidenced beneficial and synergetic effects of BC amendment on the abatement of the potential environmental risk from heavy metal polluted field on a long-term basis.


Disentangling photoperiod from hop vernalization and dormancy for global production and speed breeding.
Humulus lupulus L. (hop) flowers are a key ingredient in beer, imparting the beverage's aroma and bitterness profile. Photoperiod is known to interact with temperature to control flowering in hops. Studies have stipulated that resting dormant buds on hops require a minimum chilling duration for their meristems to break dormancy and grow fruitfully. This assertion, in part, led to a long-held notion that hops require vernalization and/or dormancy for the meristem to change from a vegetative to floral state. The research in this study aims to separate photoperiod from vernalization and dormancy through a series of experiments that artificially control photoperiod to prevent the onset of dormancy and chilling exposure. Six experiments were performed to assess flower yield and quality for seven diverse hop cultivars (with and without exposure to chilling and dormancy) to quantify the impact on flowering performance. Vernalization and dormancy, two plant traits previously considered necessary to the proliferation of hop flowers, do not influence hop flower yield and quality. The findings have broad implications; global hop production can be distributed more widely and it paves the way for speed breeding and controlled-environment production to achieve 4 hop generation cycles per year, as opposed to 1 under field-grown conditions.


Diagnostic accuracies of intraoperative frozen section and permanent section examinations for histological grades during open biopsy of bone tumors.
A histological diagnosis obtained from an intraoperative frozen section (FS) during biopsy confirms the adequacy of tumor tissue in the specimen. However, some cases show a discrepancy among the intraoperative FS diagnosis, permanent section (PS) diagnosis of the biopsy specimen, and the final diagnosis of the excised tumor specimen. In this study, we retrospectively investigated the diagnostic accuracy of the FS and PS for different types of bone tumors. This study included 377 patients with 411 bone tumors who underwent tumor excision after an open biopsy with intraoperative FS diagnosis. FS, PS, and final diagnoses of the patients were classified into benign tumors/tumor-like lesions, intermediate malignancies, and malignant tumors. To assess diagnostic accuracy, the histological grades in FS and PS diagnoses were compared with those in the final diagnoses. The overall diagnostic accuracies of FS and PS were 93% and 97%, respectively. The accuracy of FS and PS for histological grade was 84% and 93% for chondrogenic tumors, 90% and 96% for osteogenic tumors, 97% and 98% for osteoclastic giant cell-rich tumors, 100% and 100% for tumors of undefined neoplastic nature, and 95% and 99% for other bone tumors, respectively. These data suggest that surgical planning based on PS diagnosis is recommended for chondrogenic and osteogenic tumors.


Persistent challenges with treating multiple myeloma early.
Over the past decade, 2 strategies have advanced the treatment of patients with multiple myeloma and its precursor diseases. First, the definition has changed to include patients without end organ damage, who previously would not have been treated. Second, there is widespread enthusiasm for treating high-risk, smoldering multiple myeloma. In this commentary, we explore the evidence supporting these therapeutic expansions. Although early treatment adds cost and therapeutic burden, it remains unknown whether survival and health-related quality of life are improved by early treatment. Herein, we consider the implications of diagnostic expansion in multiple myeloma.


Response of the turbidity maximum zone in the Yangtze River Estuary due to human activities during the dry season.
The interaction between a river and the sea results in a turbidity maximum zone (TMZ) within the estuary, which has a great impact on the local ecosystem. In the Yangtze River Estuary, the magnitude and extent of the TMZ vary with water discharge. In this study, the cumulative human activity altered the water discharge regime from the river to the estuary. In the post-Three Gorges Dam (TGD) period, water discharge increased by 35.10 % at Datong in February compared with that in the pre-TGD period. The effects of water discharge variation on the characteristics of the TMZ were analyzed during spring and neap tidal periods using the three-dimensional environmental fluid dynamic code (EFDC) model. The area of the TMZ decreased by 3.11 and 17.39 % during neap and spring tides, respectively. In addition, the upper limit of the TMZ moved 11.68 km seaward during neap tide, whereas the upper limit of the TMZ in the upstream and downstream areas moved seaward 9.65 and 2.34 km, respectively, during spring tide. These findings suggest that the area and location of the TMZ are more sensitive to upstream runoff during spring tide than during neap tide. These changes in the TMZ will impact the biochemical processes in the Yangtze River Estuary. In the foreseeable future, the distribution characteristic of TMZ will inevitably change due to variations in the Yangtze River discharge resulting from new human activities (i.e., new dams), which are being constructed upstream in the Yangtze River system.


The paradox of anemia with high meat intake: a review of the multifactorial etiology of anemia in the Inuit of North America.
Anemia is a serious concern among indigenous populations in North America, and it appears to be widespread among the Inuit despite abundant intakes of heme iron. It is therefore hypothesized that anemia for the Inuit involves other dietary factors not usually associated with animal foods, such as low intakes of vitamin A and/or folate, riboflavin, and vitamin C. Also, Helicobacter pylori infection and/or parasitosis may result in gastrointestinal blood loss and/or functional iron deficiency. This review aims to describe factors that may cause anemia in Inuit populations despite high meat intakes, abundant bioavailable iron, and other important hematological nutrients.


AtFtsH heterocomplex-mediated degradation of apoproteins of the major light harvesting complex of photosystem II (LHCII) in response to stresses.
Chloroplastic heterocomplex consisting of AtFtsH1, 2, 5 and 8 proteases, integrally bound to thylakoid membrane was shown to play a critical role in degradation of photodamaged PsbA molecules, inherent to photosystem II (PSII) repair cycle and in plastid development. As no one thylakoid bound apoproteins besides PsbA has been identified as target for the heterocomplex-mediated degradation we investigated the significance of this protease complex in degradation of apoproteins of the major light harvesting complex of photosystem II (LHCII) in response to various stressing conditions and in stress-related changes in overall composition of LHCII trimers of PSII-enriched membranes (BBY particles). To reach this goal a combination of approaches was applied based on immunoblotting, in vitro degradation and non-denaturing isoelectrofocusing. Exposure of Arabidopsis thaliana leaves to desiccation, cold and high irradiance led to a step-wise disappearance of Lhcb1 and Lhcb2, while Lhcb3 level remained unchanged, except for high irradiance which caused significant Lhcb3 decrease. Furthermore, it was demonstrated that stress-dependent disappearance of Lhcb1-3 is a proteolytic phenomenon for which a metalloprotease is responsible. No changes in Lhcb1-3 level were observed due to exposition of var1-1 mutant leaves to the three stresses clearly pointing to the involvement of AtFtsH heterocomplex in the desiccation, cold and high irradiance-dependent degradation of Lhcb1 and Lhcb2 and in high irradiance-dependent degradation of Lhcb3. Non-denaturing isoelectrofocusing analyses revealed that AtFtsH heterocomplex-dependent differential Lhcb1-3 disappearance behaviour following desiccation stress was accompanied by modulations in abundances of individual LHCII trimers of BBY particles and that LHCII of var1-1 resisted the modulations.


Association of perioperative factor XIII activity levels and other haemostatic markers with the risk of postoperative intracranial haematoma in a selected cohort of neurosurgical patients.
Coagulation factor XIII and other haemostatic markers are known strengthen fibrin clot formation and, hence, may facilitate safer surgery. Currently however, factor XIII activity levels are not routinely screen. Therefore, the purpose of this study was to determine the association of perioperative factor XIII activity levels and other haemostatic markers with postoperative intracranial haematoma formation in neurosurgical patients. Between January 2008 to Jun 2009, all neurosurgical patients who underwent intracranial surgery were screened for the study. Patients had blood samples taken preoperatively and within 24 h post-surgery for factor XIII and other haemostatic markers. The intracranial surgeries for the patients involved were performed according to their respective indications using standard neurosurgical techniques. Postoperatively, patients had a computed tomography (CT) brain scan, with the imaging results grouped into three classes: significant haematoma (group I), insignificant haematoma (group II) and no haematoma (group III). Of the total 84 enrolled patients, 5 (6%), 28 (33.3%) and 51 (60.7%) patients were assigned to group I, II and III respectively. Significant postoperative haematoma that required re-surgery was related to low postoperative platelet count (p < 0.01), and higher odds ratio of developing postoperative intracranial haematoma were shown with two combination factors: low postoperative factor XIII and platelet levels; and low postoperative factor XIII and antithrombin levels. Low platelet count can cause significant volume postoperative intracranial haematoma and in presence of multiple defects in haemostatic markers appears to be clinically useful to predict the formation of postoperative intracranial haematoma in neurosurgical patients.


Extracerebral anaplastic meningioma.
Conventional treatments of anaplastic meningioma include surgery and radiotherapy. Temozolomide is also an option. Metastases to extracerebral organs are very rare and hardly manageable. Antiangiogenic drugs could represent a salvage therapy. Anaplastic meningioma is an unusual tumor subtype that rarely involves organs outside the brain. In our case, a patient with cerebral meningioma developed lung metastases after 19 years from the initial tumor appearance. This unpredictable behavior could make complex the diagnosis.


Comparative gene expression profile analysis of ovules provides insights into Jatropha curcas L. ovule development.
Jatropha curcas, an economically important biofuel feedstock with oil-rich seeds, has attracted considerable attention among researchers in recent years. Nevertheless, valuable information on the yield component of this plant, particularly regarding ovule development, remains scarce. In this study, transcriptome profiles of anther and ovule development were established to investigate the ovule development mechanism of J. curcas. In total, 64,325 unigenes with annotation were obtained, and 1723 differentially expressed genes (DEGs) were identified between different stages. The DEG analysis showed the participation of five transcription factor families (bHLH, WRKY, MYB, NAC and ERF), five hormone signaling pathways (auxin, gibberellic acid (GA), cytokinin, brassinosteroids (BR) and jasmonic acid (JA)), five MADS-box genes (AGAMOUS-2, AGAMOUS-1, AGL1, AGL11, and AGL14), SUP and SLK3 in ovule development. The role of GA and JA in ovule development was evident with increases in flower buds during ovule development: GA was increased approximately twofold, and JA was increased approximately sevenfold. In addition, the expression pattern analysis using qRT-PCR revealed that CRABS CLAW and AGAMOUS-2 were also involved in ovule development. The upregulation of BR signaling genes during ovule development might have been regulated by other phytohormone signaling pathways through crosstalk. This study provides a valuable framework for investigating the regulatory networks of ovule development in J. curcas.


Nonoperative Management for Rectal Cancer.
The treatment algorithm for locally advanced rectal cancer (LARC) has increased in complexity over the past 10 years. Nonoperative management (NOM) for rectal cancer in patients with clinical complete response (cCR) after neoadjuvant therapy has been gaining acceptance as a potential treatment option for selected LARC patients. The current challenge is to accurately select the patients with an apparent cCR, thereby correctly identifying those would-be appropriate candidates for a NOM strategy. NOM should be part of the treatment discussion of LARC, considering increasing rates of cCR, patient preference, potential quality of life gains, and the potential avoidance of surgical morbidity.


Excitotoxic and Radiation Stress Increase TERT Levels in the Mitochondria and Cytosol of Cerebellar Purkinje Neurons.
Telomerase reverse transcriptase (TERT) is the catalytic subunit of telomerase, an enzyme that elongates telomeres at the ends of chromosomes during DNA replication. Recently, it was shown that TERT has additional roles in cell survival, mitochondrial function, DNA repair, and Wnt signaling, all of which are unrelated to telomeres. Here, we demonstrate that TERT is enriched in Purkinje neurons, but not in the granule cells of the adult mouse cerebellum. TERT immunoreactivity in Purkinje neurons is present in the nucleus, mitochondria, and cytoplasm. Furthermore, TERT co-localizes with mitochondrial markers, and immunoblot analysis of protein extracts from isolated mitochondria and synaptosomes confirmed TERT localization in mitochondria. TERT expression in Purkinje neurons increased significantly in response to two stressors: a sub-lethal dose of X-ray radiation and exposure to a high glutamate concentration. While X-ray radiation increased TERT levels in the nucleus, glutamate exposure elevated TERT levels in mitochondria. Our findings suggest that in mature Purkinje neurons, TERT is present both in the nucleus and in mitochondria, where it may participate in adaptive responses of the neurons to excitotoxic and radiation stress.


Research note changes in the number of outpatient visits in Japan during the COVID-19 pandemic.
This study aimed to determine the change in the number of outpatient visits in Japan since the beginning of the COVID-19 pandemic, using data on the outpatient claims submitted by medical institutions to insurers in 2019 and 2020, from the National Database of Health Insurance Claims and Specific Health Check-ups. We calculated the total number of outpatient visits, and number of visits for individual diseases according to the International Classification of Diseases-10 codes on the claim form. The number of outpatient visits per million people decreased by 9.98% in 2020 compared to 2019. Of the diseases included in the analysis, 71 showed a decrease in the number of visits by ≥ 1%. There were significant decreases in the number of visits related to infectious diseases (influenza, acute bronchitis, and acute laryngitis, etc.), and chronic diseases (hemorrhoids, cystic kidney disease, dyspepsia, and chronic sinusitis, etc.). The observed decreased rate of outpatient visit might have been due to, a decrease in the incidence of disease, a decreased frequency of visit by patients with the disease, or both. Our analysis method using actual health insurance claim data can be applied worldwide, where researchers have access to national information on health insurance claims.


Imaging mass spectrometry reveals complex lipid distributions across Staphylococcus aureus biofilm layers.
Although Staphylococcus aureus is the leading cause of biofilm-related infections, the lipidomic distributions within these biofilms is poorly understood. Here, lipidomic mapping of S. aureus biofilm cross-sections was performed to investigate heterogeneity between horizontal biofilm layers. S. aureus biofilms were grown statically, embedded in a mixture of carboxymethylcellulose/gelatin, and prepared for downstream matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI IMS). Trapped ion mobility spectrometry (TIMS) was also applied prior to mass analysis. Implementation of TIMS led to a ∼ threefold increase in the number of lipid species detected. Washing biofilm samples with ammonium formate (150 mM) increased signal intensity for some bacterial lipids by as much as tenfold, with minimal disruption of the biofilm structure. MALDI TIMS IMS revealed that most lipids localize primarily to a single biofilm layer, and species from the same lipid class such as cardiolipins CL(57:0) - CL(66:0) display starkly different localizations, exhibiting between 1.5 and 6.3-fold intensity differences between layers (n = 3, p < 0.03). No horizontal layers were observed within biofilms grown anaerobically, and lipids were distributed homogenously. High spatial resolution analysis of S. aureus biofilm cross-sections by MALDI TIMS IMS revealed stark lipidomic heterogeneity between horizontal S. aureus biofilm layers demonstrating that each layer was molecularly distinct. Finally, this workflow uncovered an absence of layers in biofilms grown under anaerobic conditions, possibly indicating that oxygen contributes to the observed heterogeneity under aerobic conditions. Future applications of this workflow to study spatially localized molecular responses to antimicrobials could provide new therapeutic strategies.


Proteogenomic characterization of 2002 human cancers reveals pan-cancer molecular subtypes and associated pathways.
Mass-spectrometry-based proteomic data on human tumors-combined with corresponding multi-omics data-present opportunities for systematic and pan-cancer proteogenomic analyses. Here, we assemble a compendium dataset of proteomics data of 2002 primary tumors from 14 cancer types and 17 studies. Protein expression of genes broadly correlates with corresponding mRNA levels or copy number alterations (CNAs) across tumors, but with notable exceptions. Based on unsupervised clustering, tumors separate into 11 distinct proteome-based subtypes spanning multiple tissue-based cancer types. Two subtypes are enriched for brain tumors, one subtype associating with MYC, Wnt, and Hippo pathways and high CNA burden, and another subtype associating with metabolic pathways and low CNA burden. Somatic alteration of genes in a pathway associates with higher pathway activity as inferred by proteome or transcriptome data. A substantial fraction of cancers shows high MYC pathway activity without MYC copy gain but with mutations in genes with noncanonical roles in MYC. Our proteogenomics survey reveals the interplay between genome and proteome across tumor lineages.


α-Crystallins Are Small Heat Shock Proteins: Functional and Structural Properties.
During its life cycle, a cell can be subjected to various external negative effects. Many proteins provide cell protection, including small heat shock proteins (sHsp) that have chaperone-like activity. These proteins have several important functions involving prevention of apoptosis and retention of cytoskeletal integrity; also, sHsp take part in the recovery of enzyme activity. The action mechanism of sHsp is based on the binding of hydrophobic regions exposed to the surface of a molten globule. α-Crystallins presented in chordate cells as two αA- and αB-isoforms are the most studied small heat shock proteins. In this review, we describe the main functions of α-crystallins, features of their secondary and tertiary structures, and examples of their partners in protein-protein interactions.


Hormonal stress response and hemodynamic stability in patients undergoing endovascular vs. conventional abdominal aortic aneurysm repair.
To investigate the effect of two different surgical techniques with different anesthetic modes on intraoperative and postoperative hormonal stress response, hemodynamic stability, fluid loading and renal function in patients scheduled for elective infrarenal abdominal aortic aneurysm (AAA) repair. Forty consecutive patients scheduled for elective infrarenal AAA repair were allocated without randomizing into two groups: an endovascular (EVAR, n = 20) and a conventional (CAR, n = 20) aneurysm repair group according to aneurysm morphology as determined by preoperative computed tomography and angiography. The EVAR group were operated under spinal anesthesia and the CAR group using general anesthesia with epidural blockade. Patients undergoing CAR showed lower intraoperative mean arterial pressure and significantly higher plasma norepinephrine before aortic cross-clamping and significantly higher lactate after aortic declamping and postoperatively than patients in the EVAR group. Postoperatively, vasopressin and serum cortisol were also significantly higher in the CAR group. Fluid loading and estimated blood loss were more excessive in the CAR group. Stress response was lower and hemodynamic stability and lower body perfusion superior and renal function also better maintained in patients undergoing EVAR under spinal anesthesia as compared to those undergoing CAR using general anesthesia with epidural blockade.


Assessment of the efficacy and safety of platelet poor plasma gel as autologous dermal filler for facial rejuvenation.
The popularity of dermal fillers has grown rapidly in the last few years for facial rejuvenation. Plasma gel is an autologous gelatinous material that is prepared from the patient's own platelet poor plasma. To evaluate the clinical efficacy and safety of plasma gel injection as a dermal filler for facial rejuvenation. The current study was carried out on 52 females presented with facial aging divided into two groups: Group A included 34 females with facial wrinkles, and Group B included 18 females with tear trough deformity. After taking written informed consent, they received two sessions of plasma gel injections at 2-week interval and followed up monthly for 3 months. They were assessed clinically before treatment sessions and at the end of follow-up period. Both studied groups showed immediate significant clinical improvement after plasma gel injection that maintained till the end of follow-up period. This finding was confirmed by significant reduction in the mean values of Wrinkle Severity Rating Scale (WSRS) in Group A and Tear Trough Rating Scale (TTRS) in group B, and significant improvement of skin homogeneity and texture in both groups. In general, the reported side effects were minimal and transient. Autologous platelet poor plasma gel injection seems to be a cost-effective, safe, well-tolerated, and minimally invasive technique producing significant aesthetic correction of facial wrinkles and tear trough deformity.


Climate and human mortality in Virginia, 2005-2020.
Using an extensive database of every resident death in Virginia from 2005 to 2020, climate-mortality relationships are examined for 12 climatically homogeneous regions within the Commonwealth. Each region is represented by a first-order weather station from which archived temperature and humidity data are used to generate a variety of biometeorologically relevant indices. Using these indices and other variables (such as air quality and heat and cold waves), daily mortality and climate relationships are modeled for each region over a 21-day lag period utilizing generalized additive models and distributed lag non-linear models. Optimal models are identified for each region, and a consensus model was also run based on maximum temperature to facilitate inter-regional comparisons. The relative risk of mortality varies markedly as a function of climate between regions, with U-shaped, J-shaped, and inverse linear relationships evident. Cold mortality exceeds heat mortality across most of Virginia (typical relative risks are 1.10 for cold and 1.03 for heat), with cold risks strongest at lags 3 to 10. Low temperatures (or low humidity) are protective at lags 0-2 days except in the colder, western parts of state. Heat mortality occurs at short lags (0-2 days) for three-fourths of the stations, but the spatial pattern is random. Mortality displacement is evident for most regions for several days following the heat-related spike. Although the use of region-specific models is justified, the simple consensus model based on a consistent set of predictors provides similar results.


B7 family checkpoint regulators in immune regulation and disease.
Fine-tuning the immune response and maintaining tolerance to self-antigens involves a complex network of co-stimulatory and co-inhibitory molecules. The recent FDA approval of ipilimumab, a monoclonal antibody blocking cytotoxic T lymphocyte antigen (CTLA)-4, demonstrates the impact of checkpoint regulators in disease. This is reinforced by ongoing clinical trials targeting not only CTLA-4, but also the programmed death (PD)-1 and B7-H4 pathways in various disease states. Recently, two new B7 family inhibitory ligands, V-domain Ig suppressor of T cell activation (VISTA) and B7-H6 were identified. Here, we review recent understanding of B7 family members and their concerted regulation of the immune response to either self or foreign pathogens. We also discuss clinical developments in targeting these pathways in different disease settings, and introduce VISTA as a putative therapeutic target.


Peripheral nerve involvement in hereditary spastic paraplegia characterized by quantitative magnetic resonance neurography.
Hereditary spastic paraplegias (HSPs) are heterogenous genetic disorders. While peripheral nerve involvement is frequent in spastic paraplegia 7 (SPG7), the evidence of peripheral nerve involvement in SPG4 is more controversial. We aimed to characterize lower extremity peripheral nerve involvement in SPG4 and SPG7 by quantitative magnetic resonance neurography (MRN). Twenty-six HSP patients carrying either the SPG4 or SPG7 mutation and 26 age-/sex-matched healthy controls prospectively underwent high-resolution MRN with large coverage of the sciatic and tibial nerve. Dual-echo turbo-spin-echo sequences with spectral fat-saturation were utilized for T2-relaxometry and morphometric quantification, while two gradient-echo sequences with and without an off-resonance saturation rapid frequency pulse were applied for magnetization transfer contrast (MTC) imaging. HSP patients additionally underwent detailed neurologic and electroneurographic assessments. All microstructural (proton spin density [ρ], T2-relaxation time, magnetization transfer ratio) and morphometric (cross-sectional area) quantitative MRN markers were decreased in SPG4 and SPG7 indicating chronic axonopathy. ρ was superior in differentiating subgroups and identifying subclinical nerve damage in SPG4 and SPG7 without neurophysiologic signs of polyneuropathy. MRN markers correlated well with clinical scores and electroneurographic results. MRN characterizes peripheral nerve involvement in SPG4 and SPG7 as a neuropathy with predominant axonal loss. Evidence of peripheral nerve involvement in SPG4 and SPG7, even without electroneurographically manifest polyneuropathy, and the good correlation of MRN markers with clinical measures of disease progression, challenge the traditional view of the existence of HSPs with isolated pyramidal signs and suggest MRN markers as potential progression biomarkers in HSP.


Electron microscopic determination of silver incorporation in collagen fibers as a model of organic-metal chiral supramolecular structure with optical anisotropic properties.
The high positive linear dichroism (LD) exhibited by silver-impregnated collagen fibers are thought to be caused by an ordered binding of silver nanoparticles to the collagen fibers. A correlation between LD, established by image analysis, and the size of silver particles as measured by electron microscopy was investigated in the same sections of silver-impregnated tendon collagen fibers. Silver particles were found to be periodically ordered along the collagen fibers, with some preference for the 67 nm collagen period and collagen a bands. The silver particle diameter for a sample of n=279 was 18.51 nm on average, with 95% of the values ranging between 17.90 and 19.14 nm. The finding that interference colors are correlated to abnormal dispersion of birefringence characteristics supports the concept that collagen fibers induce an ordered binding of silver distributed along the fibers.


Magnetic resonance spectroscopy for quantification of liver iron deposition in hereditary hemochromatosis of a Chinese family: Four case reports.
Hereditary hemochromatosis (HH) is a major cause of liver iron overload. The gold standard for the diagnosis of liver iron overload is the histopathological analysis of a liver sample collected by biopsy. The biopsy procedure is both invasive and painful and carries some risks of complications. The multi-echo single-voxel magnetic resonance spectroscopy (HISTO) technique can be used for noninvasive, quantitative assessment of liver iron overload. We report 4 Chinese Han men, who were relatives. Patient A was admitted with diabetes and presented with thrombocytopenia and skin hyperpigmentation. The other patients had no specific clinical presentation. Patient A was suspected of having iron in the liver on routine magnetic resonance imaging, therefore, further HISTO, laboratory testing, and liver biopsy were performed, which confirmed iron metabolic abnormalities. Furthermore, we identified hepatic iron deposition using HISTO and laboratory testing of his son and 2 brothers. Combined with symptoms, auxiliary examinations, and liver biopsy, HH was considered. As the 4 patients had no other discomfort other than patient A who had diabetes, patient A was placed on therapy comprising the insulin pump, acarbose, and platelet booster capsule. After treatment, the diabetic symptoms of patient A improved. The patient and his relatives were regularly followed-up for HH. HH should be considered when hepatic iron deposition is suspected by routine magnetic resonance, as the HISTO sequence can quantitate liver iron deposition and leads to a promising diagnosis. HISTO is of great value in familial cases, especially in young patients requiring long-term follow-up.


Bilateral Achilles tendon rupture: a case report.
Bilateral Achilles tendon rupture is a rare injury. We present a case of a 59-year-old gentleman who sustained a bilateral Achilles tendon rupture when the tendon was subjected to normal physiological load. He was treated operatively with V-Y plasty and repair of the tendon with post-operative plaster immobilisation.


Highly sensitive determination of diclofenac based on resin beads and a novel polyclonal antibody by using flow injection chemiluminescence competitive immunoassay.
A novel flow injection chemiluminescence immunoassay for simple, sensitive and low-cost detection of diclofenac was established based on specific binding of antigen and antibody. Carboxylic resin beads used as solid phase carrier materials provided good biocompatibility and large surface-to-volume ratio for modifying more coating antigen. There was a competitive process between the diclofenac in solution and the immobilized coating antigen to react with the limited binding sites of the polyclonal antibody to form the immunocomplex. The second antibody labelled with horseradish peroxidase was introduced into the immunosensor and trapped by captured polyclonal antibody against diclofenac, which could effectively amplify chemiluminescence signals of luminol-PIP-H2O2. Under optimal conditions, the diclofenac could be detected quantitatively. The chemiluminescence intensity decreased linearly with the logarithm of the diclofenac concentration in the range of 0.1-100ngmL-1 with a detection limit of 0.05ngmL-1 at a signal-to-noise ratio of 3. The immunosensor exhibited high sensitivity, specificity and acceptable stability. This easy-operated and cost-effective analytical method could be valuable for the diclofenac determination in real water samples.


Effectiveness of Platelet-Rich Plasma in the Treatment of Androgenic Alopecia: A Meta-Analysis.
Androgenetic alopecia (AGA) is a common yet difficult-to-treat condition, which is an important psychosocial problem. Platelet-rich plasma (PRP) therapy has been considered as a promising treatment for AGA. However, the current evidence on the efficacy of PRP for treating AGA is still controversial. This study evaluated the efficacy of PRP monotherapy in the treatment of AGA. We searched PubMed, Embase, Cochrane Library and Web of Science to collect randomized controlled trials on use of PRP in AGA for a meta-analysis. Ten trials with a total 555 treatment units were identified. The hair density in PRP group was significantly higher than control group [MD = 25.09, 95%CI: 9.03-41.15, p = 0.002], but there was no significant difference in hair diameter between two groups [SMD = 0.57, 95%CI: - 0.23 to 1.38, p = 0.16]. Subgroup analyses indicated that hair density was significantly higher among the male-only trials than in the mixed-sex samples (p = 0.02). In addition, neither the split-head design nor the year of publication affected hair density (p = 0.05, p = 0.06). However, hair density was significantly higher in trials with a sample size less than 30 (p = 0.0004). PRP treatment increased hair density in participants with AGA, but not hair diameter. In terms of hair density, PRP elicits stronger effects in male patients. There was a trend toward differed treatment effect by gender with PRP injection, which warrants further investigation. Especially in the case of female. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors https://www.springer.com/00266 .


Cocoa butter symmetrical monounsaturated triacylglycerols: separation by solvent fractionation and application as crystallization modifier.
Symmetrical monounsaturated triacylglycerols-also known as SUS (saturated unsaturated saturated triacylglycerols)-are the main triacylglycerols present in cocoa butter (CB), exhibiting decisive influence in the tempering process conditions and sensorial characteristics in chocolate products. CBs with lower amounts of SUS do not crystallize adequately in chocolate processing, generating a low quality product with higher susceptibility to fat bloom formation. In this context, fractions with higher contents of SUS triacylglycerols were obtained by solvent fractionation of cocoa butter, under four operating conditions varying crystallization temperatures (17, 18, 20 and 22 °C) and sample/acetone solvent ratios (1:7 and 1:5, in w/v). After solvent evaporation, CB fractions were evaluated in terms of triacylglycerol profile, solid fat content and melting point. The solvent fractionation performed at 17 °C with a 1:7 ratio of cocoa butter:acetone favored the separation of a higher concentrated SUS stearin (denominated as S17) with 7.1% increment in SUS triacylglycerols, mainly StOSt-1-stearyl-2-oleoyl-stearin (26.9%) and POSt-1-palmitoyl-2-oleoyl-stearin (45.0%). Additionally, S17 was added to pure cocoa butter at three different proportions of CB:S17 (95:5, 90:10 and 80:20, in w/w) and the blend was evaluated by isothermal crystallization (at 17.5 °C) and consistency (at 10, 15 and 20 °C). Compared to the pure cocoa butter, at 10 °C, the blend with 20% stearin (80:20) showed an increment of 67% in the consistency and considerable higher crystallization rates at 17.5 °C were found, confirming the potential contribution of the SUS triacylglycerols as crystallization modifiers of CB.


Do women live longer following lung resection for carcinoma?
To determine whether patient gender affects the outlook following lung resection for non-small cell lung cancer (NSCLC). Prospectively collected data on 833 patients undergoing lung resection for NSCLC between 1990 and 2000 in a single unit were analysed. 581 patients were male (mean age 64.7 +/- 7 years) and 252 were female (mean age 62.6 +/- 7.8 years) (P=0.006). Male patients were more likely to have a history of ischaemic heart disease (P=0.03), to have poorer preoperative spirometry as demonstrated by their % predicted FEV1 (P=0.02) and to need pneumonectomy (P=0.0001) than their female counterparts. Squamous cell carcinoma was the predominant histological cell type in men and adenocarcinoma in women (P<0.0001). There was a trend towards a lower pathological stage among women, but this was not significant. Operative mortality for men was 4.6 and 1.2% for women (P=0.01). Overall 5-year survival for men was 34.2 +/- 2.65% and 47.5 +/ - 4.2% for women (P=0.001) and, for the hospital survivors, was 36.5 +/- 2.7% and 48.1 +/- 4.2%, respectively (P=0.01). On univariate analysis, older age, the need for pneumonectomy and higher pathological stage were significant adverse factors whereas squamous cell type and female gender were significant favourable factors for survival (P<0.05). On Cox proportional hazards model (with and without hospital deaths), pathological stage (P<0.0001), female gender (P=0.0006) and squamous cell type (P=0.001) were independent predictors of survival. The survival was significantly better for women having squamous cell (P=0.01) or non-squamous cell cancers (adenocarcinoma and other) (P=0.002). Regarding the stage, women had a significant survival advantage at pathological stage I (P=0.01) and a relatively better survival at stage II and stage III disease (P=0.3). This study suggests that female gender exerts a significant positive effect on survival following lung resection for NSCLC. This effect is pronounced at early disease stage and persists after adjusting for important differences in the clinical, histo-pathological features and extent of pulmonary resection between male and female patients.


Active translocation of platelets into sinusoidal and Disse spaces in the liver in response to lipopolysaccharides, interleukin-1 and tumor necrosis factor.
1. Injection of lipopolysaccharides (LPS) or endotoxin into mice and rats induces a prolonged increase in serotonin (5-hydroxytryptamine: 5HT), predominantly in the liver. 2. The 5HT increase reflects the accumulation of platelets in the sinusoidal and perisinusoidal Disse spaces (spaces between endothelial cells and hepatocytes) in the liver. 3. Most of the platelets which accumulated in these spaces still retained their intact structure and a large amount of 5HT. 4. Interleukin-1 and/or tumor necrosis factor also induce the platelet response. 5. Kupffer's cells play a key role in this platelet response. 6. Anti-platelet drugs currently used, except for anti-inflammatory steroids, were ineffective in preventing the platelet response. 7. This platelet response is different from the well known platelet aggregation. 8. The possible involvement of this platelet response in insulin-independent hypoglycaemia, disseminated intravascular coagulation, septic shock, hepatitis, Shwartzman type reactions or self-defense mechanisms is discussed.


The ancestry of allergy: being an account of the original experimental induction of hypersensitivity recognizing the contribution of Paul Portier.
The Nobel Prize in Medicine/Physiology for 1913 was awarded to Charles Richet, French physiologist, for studies of "anaphylaxis." The earliest articles on what was actually the original experimental induction of hypersensitivity were the result of collaborative studies of Paul Portier and Charles Richet. This article presents an account of the discovery based on an extensive examination of original sources with attention to the particulars of the contributions of each of the codiscoverers. An unusually complete and intimate story of the unfolding of the discovery could be pieced together from the findings in the original sources. The contributions of Paul Portier in the experiments leading to the recognition of a relationship between hypersensitivity and immunity earned him a more prominent position in the ancestry of allergy.


The change of plasma galectin-3 concentrations after traumatic brain injury.
Galectin-3 plays a significant role in microglia activation. Its increased circulating concentration has been associated with some inflammatory diseases. In-hospital major adverse events (IMAEs), including acute traumatic coagulopathy, progressive hemorrhagic injury and posttraumatic cerebral infarction, have high prevalence and are strong predictors of mortality after severe traumatic brain injury (STBI). The present study was designed to investigate the relationships between plasma galectin-3 concentrations and trauma severity, in-hospital mortality and IMAEs following STBI. Plasma galectin-3 concentrations of 100 STBI patients and 100 controls were determined. Diagnosis of progressive hemorrhagic injury and posttraumatic cerebral infarction was made on the follow-up computerized tomography scan. Acute traumatic coagulopathy was defined based on coagulation test. Plasma galectin-3 concentrations were significantly higher in patients as compared to controls and also associated highly with Glasgow Coma Scale scores and plasma C-reactive protein concentrations. Galectin-3 emerged as an independent predictor for in-hospital mortality and IMAEs. Areas under receiver operating characteristic curve of plasma galectin-3 concentrations were similar to those of Glasgow Coma Scale scores for prediction of in-hospital morality and IMAEs. Plasma galectin-3 concentrations have close relation to inflammation, trauma severity and clinical outcome, suggesting that galectin-3 should have the potential to be a good prognostic biomarker after STBI.


Variance associated with the use of relative velocity for force platform gait analysis in a heterogeneous population of clinically normal dogs.
Factors that contribute to variance in ground reaction forces (GRFs) include dog morphology, velocity, and trial repetition. Narrow velocity ranges are recommended to minimize variance. In a heterogeneous population, it may be preferable to minimize data variance and efficiently perform force platform gait analysis by evaluation of each individual dog at its preferred velocity, such that dogs are studied at a similar relative velocity (V*). Data from 27 normal dogs were obtained including withers and shoulder height. Each dog was trotted across a force platform at its preferred velocity, with controlled acceleration (±0.5 m/s(2)). V* ranges were created for withers and shoulder height. Variance effects from 12 trotting velocity ranges and associated V* ranges were examined using repeated-measures analysis-of-covariance. Mean bodyweight was 24.4 ± 7.4 kg. Individual dog, velocity, and V* significantly influenced GRF (P <0.001). Trial number significantly influenced thoracic limb peak vertical force (PVF) (P <0.001). Limb effects were not significant. The magnitude of variance effects was greatest for the dog effect. Withers height V* was associated with small GRF variance. Narrow velocity ranges typically captured a smaller percentage of trials and were not consistently associated with lower variance. The withers height V* range of 0.6-1.05 captured the largest proportion of trials (95.9 ± 5.9%) with no significant effects on PVF and vertical impulse. The use of individual velocity ranges derived from a withers height V* range of 0.6-1.05 will account for population heterogeneity while minimizing exacerbation of lameness in clinical trials studying lame dogs by efficient capture of valid trials.


Lab-on-chip device for single cell trapping and analysis.
Traditional cell assay gives us an average result of multiple cells and it is assumed that the resultant is the outcome of all cells in population. However, single cell studies have revealed that individual cells of same type may differ dramatically and these differences may have important role to play in cells functionality. Such information can be obscured in only studying cell population experimental approach. To uncover biological principles and ultimately to improve the detection and treatment of disease, new approaches are highly required to single cell analysis. We propose to fabricate a lab on chip device to study high throughput single cell nanotoxicity analysis. The chip incorporates independently addressable active microwell electrodes for cell manipulation and analysis. We employed positive-dielectrophoresis approach to quickly and efficiently capture single cells in each wells with having control over individual microwells. We examined change in impedance properties to verify cell capture in microwell and its health and present a novel model of single cell assay for nanotoxicity, and drug testing.


Reconstruction of rat skeletal muscle glycerophosphate shuttle.
The activity of alpha-glycerophosphate shuttle in homogenate and in a reconstructed system (isolated mitochondria and cytoplasm) from rat skeletal muscle is presented. The influence of some inhibitors of mitochondrial alpha-glycerosphate dehydrogenase on the activity alpha-glycerophosphate shuttle is demonstrated. The possible operativity of alpha-glycerophosphate shuttle in skeletal muscle in vivo is discussed.


Solid-supported lipid multilayers: structure factor and fluctuations.
We present a theoretical description of the thermal fluctuations in a solid-supported stack of lipid bilayers, for the case of vanishing surface tension gamma = 0 and in the framework of continuous smectic elasticity. The model is successfully used to model the reflectivity profile of a thin (16 bilayers) DMPC sample under applied osmotic pressure and the diffuse scattering from a thick (800 bilayers) stack. We compare our model to previously existing theories.


A new species of freshwater crab of the genus Kingsleya Ortmann, 1897 (Crustacea: Brachyura: Pseudothelphusidae) from Amazonia, Brazil.
A new species of freshwater crab, Kingsleya celioi, from the Brazilian Amazon (Pará State) is described and illustrated. The new species can be easily separated from their congeners by a suite of morphological characters, including the apical plate of the first gonopod large, widest medially in abdominal view, with single large spine-like outgrowth in midlength of mesial margin; distal, proximal lobes of apical plate unequal in size, distal lobe largest, tapering distally in lateral view.


Synthesis of novel structurally simplified estrogen analogues.
A library of 17 novel estrogen analogues 3 and 4 containing different substituents at rings A and D (steroid nomenclature) was prepared in a five- to seven-step synthesis. The key transformation is a Sonogashira-coupling of cyclic vinyl iodides of type 7 or 8 with phenylacetylenes of type 9. Reduction of the keto function in 3 led to the estradiol analogue 5.


Kinetics of the intestinal brush border proline (Imino) carrier.
The kinetics of L-proline transport across intestinal brush borders via the Imino carrier were studied using membrane vesicles. The Imino carrier is defined as the agent responsible for L-alanine insensitive. Na+-dependent uptake of L-proline. Initial rate measurements were made under voltage clamped conditions (pD = 0) to investigate L-proline transport as a function of cis and trans Na+ and proline concentrations. Under zero-trans conditions, increasing cis Na+ activated proline uptake with a Hill coefficient of 1.7 and decreased the apparent Kt with no change in Jimax. The Jimax was approximately 60 pmol mg-1 s-1 and the apparent Kt ranged from 0.25 mM at cis Na = 100 to 1.0 mM at cis Na+ = 30 mM. Trans Na inhibited proline uptake via a reduction in Jimax. Trans proline had no significant effect in the absence of trans Na+, but it relieved the trans Na+ inhibition. Under equilibrium exchange conditions, the Jimax was twice that observed under zero-trans conditions. These kinetics of L-proline transport suggest a model in which uptake occurs by a rapid equilibrium iso-ordered ter ter system. Two Na+ ions bind first to the carrier on the cis face of the membrane to increase the affinity of the carrier for proline. The fully loaded complex then isomerizes to release the substrates to the trans side. The partially loaded Na+-only forms are unable to translocate across the membrane. A rate-limiting step appears to be the isomerization of unloaded carrier from the trans to the cis side of the membrane.


Spinous Process Splitting Laminectomy for Lumbar Spinal Stenosis: 2D Operative Video.
Degenerative lumbar spinal stenosis involves an acquired reduction in the spinal canal diameter due to osteoarthritic changes on the disk, facet joints, and ligaments and may result in spinal cord or cauda equina compression.1 This process may lead to pain radiating to the legs, neurogenic claudication, and neurologic deficit. First-line treatment includes conservative care such as physical therapy, spinal injections, and lifestyle changes. If this strategy is insufficient to achieve symptom relief, surgical management is recommended.1,2 Surgery generally encompasses a decompression procedure through a posterior approach. There are several techniques to accomplish this in the context of severe bilateral stenosis including standard open laminectomy, unilateral laminectomy with bilateral decompression, and a tubular approach with bilateral decompression (e.g., "over-the-top technique").2 Among these, the spinous process splitting laminectomy has emerged as a strategy that allows decompressing the spinal canal through a familiar anatomy to the surgeon while respecting paravertebral muscles.3,4 This technique involves exposure of the laminae by cutting through the spinous process and then separating both halves and muscles attached at the sides. The main advantage is that the insertion of these paravertebral soft tissues is preserved, the required retraction is reduced and postoperative pain is decreased.4 Moreover, the learning curve to achieve a successful decompression employing the splitting laminectomy is substantially shorter than with other minimally invasive approaches, such as tubular. This video aims to show the steps to perform this technique (Video 1). We report the case of a 74-year-old male who presented with left sciatica and neurogenic claudication. The images showed multilevel degenerative lumbar spinal stenosis, with severe bilateral compression at L4-5, without signs of instability. Surgical alternatives were discussed with the patient, and it was decided to perform an L4-5 spinous process splitting laminectomy. The patient had a good evolution with an unremarkable postoperative course.


Hospital-specific probability of cystectomy affects survival from muscle-invasive bladder cancer.
Radical cystectomies (RCs) are increasingly centralized, but bladder cancer can be diagnosed in every hospital The aim of this study is to assess the variation between hospitals of diagnosis in a patient's chance to undergo a RC before and after the volume criteria for RCs, to identify factors associated with this variation and to assess its effect on survival. Patients diagnosed with muscle-invasive bladder cancer (cT2-4a,N0/X,M0/X) without nodal or distant metastases between 2008 and 2016 were identified through the Netherlands Cancer Registry. Multilevel logistic regression analysis was used to investigate the hospital specific probability of undergoing a cystectomy. Cox proportional hazard regression analysis was used to assess the case-mix adjusted effect of hospital-specific probabilities on survival. Of the 9,215 included patients, 4,513 (49%) underwent a RC. The percentage of RCs varied between 7% and 83% by hospital of diagnosis before the introduction of the first volume criteria (i.e., 2008-2009; minimum of 10 RCs). This variation decreased slightly to 17%-77% after establishment of the second volume criteria (i.e., 2015-2016; minimum of 20 RCs). Age, cT-stage and comorbidity were inversely and socioeconomic status was positively associated with RC. Both being diagnosed in a community hospital and/or being diagnosed in a hospital fulfilling the RC volume criteria were associated with increased use of RC compared to academic hospitals and hospitals not fulfilling the volume criteria. For each 10% increase in the percentage of RC in the hospital of diagnosis, 2-year case-mix adjusted survival increased 4% (hazard ratio 0.96, 95% confidence interval 0.94-0.98). Probability of RC varied between hospitals of diagnosis and affected 2-year overall survival. Undergoing a RC was associated with age, cT-stage, socioeconomic status, type of hospital, and whether the hospital of diagnosis fulfilled the RC volume criteria. Future research is needed to identify patient, tumor, and hospital characteristics affecting utilization of curative treatment as this may benefit overall survival.


Synthesis of cell-penetrating peptides for cargo delivery.
Cell-penetrating peptides (CPPs) have served as vehicles for the delivery of different molecules and particles into cells. The efficiency and noninvasive nature of peptide-mediated cellular transduction provides a promising tool for biomedical research. Studies of cell-penetrating constructs that deliver oligonucleotides, peptides, and proteins have elucidated several important cellular signaling mechanisms. Here we briefly describe the major classes of cell-penetrating peptides. We also present various strategies used to couple different cargoes to CPPs.