Abnormal mitosis triggers p53-dependent cell cycle arrest in human tetraploid cells.
Erroneously arising tetraploid mammalian cells are chromosomally instable and may facilitate cell transformation. An increasing body of evidence shows that the propagation of mammalian tetraploid cells is limited by a p53-dependent arrest. The trigger of this arrest has not been identified so far. Here we show by live cell imaging of tetraploid cells generated by an induced cytokinesis failure that most tetraploids arrest and die in a p53-dependent manner after the first tetraploid mitosis. Furthermore, we found that the main trigger is a mitotic defect, in particular, chromosome missegregation during bipolar mitosis or spindle multipolarity. Both a transient multipolar spindle followed by efficient clustering in anaphase as well as a multipolar spindle followed by multipolar mitosis inhibited subsequent proliferation to a similar degree. We found that the tetraploid cells did not accumulate double-strand breaks that could cause the cell cycle arrest after tetraploid mitosis. In contrast, tetraploid cells showed increased levels of oxidative DNA damage coinciding with the p53 activation. To further elucidate the pathways involved in the proliferation control of tetraploid cells, we knocked down specific kinases that had been previously linked to the cell cycle arrest and p53 phosphorylation. Our results suggest that the checkpoint kinase ATM phosphorylates p53 in tetraploid cells after abnormal mitosis and thus contributes to proliferation control of human aberrantly arising tetraploids.


In-hospital Outcomes of Attempting More Than One Chronic Total Coronary Occlusion Through Percutaneous Intervention During the Same Procedure.
The frequency and outcomes of patients who underwent chronic total occlusion (CTO) percutaneous coronary intervention (PCI) of more than one CTO during the same procedure have received limited study. We compared the clinical and angiographic characteristics and procedural outcomes of patients who underwent treatment of single versus >1 CTOs during the same procedure in 20 centers from the United States, Europe, and Russia. A total of 2,955 patients were included: mean age was 65 ± 10 years and 85% were men with high prevalence of previous myocardial infarction (46%), and previous coronary artery bypass graft surgery (33%). More than one CTO lesions were attempted during the same procedure in 58 patients (2.0%) and 70% of them were located in different major epicardial arteries. Compared with patients who underwent PCI of a single CTO, those who underwent PCI of >1 CTOs during the same procedure had similar J-CTO (2.4 ± 1.3 vs 2.5 ± 1.3, p = 0.579) and Prospective Global Registry for the Study of Chronic Total Occlusion Intervention (1.5 ± 1.2 vs 1.3 ± 1.0 p = 0.147) scores. The multi-CTO PCI group had similar technical success (86% vs 87%, p = 0.633), but higher risk of in-hospital major complications (10.3% vs 2.7%, p = 0.005), and consequently numerically lower procedural success (79% vs 85%, p = 0.197). The multi-CTO PCI group had higher in-hospital mortality (5.2% vs 0.5%, p = 0.005) and stroke (5.2%vs 0.2%, p <0.001), longer procedure duration (162 [117 to 242] vs 122 [80 to 186] minutes, p <0.001) and higher radiation dose (3.6 [2.1 to 6.4] vs 2.9 [1.7 to 4.7] Gray, p = 0.033). In conclusion, staged revascularization may be the preferred approach in patients with >1 CTO lesions requiring revascularization, as treatment during a single procedure was associated with higher risk for periprocedural complications.


Diagnosis of myasthenia gravis: Comparison of anti-nicotinic acetyl choline receptor antibodies, repetitive nerve stimulation and Neostigmine tests at a tertiary neuro care centre in India, a ten year study.
Anti-nicotinic AChR antibodies (Anti-nAChR antibodies), Repetitive Nerve Stimulation (RNS) and Neostigmine test are used for diagnosis of myasthenia gravis (MG). We compared their diagnostic agreement in a cohort of 486 MG patients over a period of ten years. Anti-nAChR antibodies, RNS and Neostigmine test showed positivity of 57.36%, 51.78%, and 93.4% respectively in ocular myasthenia and 93.77%, 82.35%, and 97.92% respectively in generalized myasthenia group. Neostigmine test showed higher positivity than anti-nAChR antibodies and RNS test in both groups. A marginal to fair agreement was observed between these tests highlighting their significance in the diagnosis of the disease.


Vocal Outcomes Following Laser-Assisted Sulcus Release for Superficial and Deep Vocal Fold Sulcus.
The objective is to study vocal outcomes following laser-assisted sulcus release (LASR), having documented the length and depth of the sulcus intraoperatively. LASR performed for superficial and deep sulci were included. Retrospective, observational. The preoperative and 3-month postoperative and post voice-therapy Voice Handicap Index 10 (VHI), Grade-Roughness-Breathiness-Asthenia-Strain of the voice (GRBAS), Maximum Phonation Time (MPT), Fundamental Frequency (FO), and stroboscopy findings were noted retrospectively from our voice clinic records of LASR patients. Intraoperative sulci length and depth details were noted from records. Patients with coexisting lesions, surgically tackled, were excluded. In a total of 14 patients, 21 superficial sulci and eight deep sulci were operated by LASR. The average preoperative VHI improved from 25.3 to 9.9, GRBAS from 10.6 to 3.5, and MPT from 7.6 to 12.3 seconds. The preoperative average FO was 235.9 Hz in 12 patients (no pick-up in two) with a postoperative 248.4 Hz. Using the Mann-Whitney U test, we found a statistically significant improvement in MPT, VHI, and GRBAS of the patients. There was no significant improvement in the FO of the patients. There was no postoperative voice deterioration in any patient. A preoperative phonatory gap was found in all patients, which postoperatively was absent in 10, markedly decreased in three, present in one with stroboscopy revealing an improvement in mucosal wave amplitude in 13 patients. In four patients with deep bilateral sulci where the LASR had been ligament deep, VHI improved from 26 to 13.3, GRBAS from 12 to 6.5, and MPT from 4.3 to 9.5 seconds. Using the Mann-Whitney U test, all three parameters were significantly improved. A statistically significant vocal improvement in VHI, GRBAS, and MPT was observed at 3 months, in superficial and deep sulci, operated by LASR. LASR is a simple, quick, and easily replicable surgery. Larger multi-centric studies with long-term follow-up are recommended.


Quadrupole mass filter: design and performance for operation in stability zone 3.
The predicted performance of a quadrupole mass filter (QMF) operating in Mathieu stability zone 3 is described in detail using computer simulations. The investigation considers the factors that limit the ultimate maximum resolution (Rmax) and percentage transmission (%Tx), which can be obtained for a given QMF for a particular scan line of operation. The performance curve (i.e., the resolution (R) versus number (N) of radio frequency (rf) cycles experienced by the ions in the mass filter) has been modeled for the upper and lower tip of stability zone 3. The saturation behavior of the performance curve observed in practice for zone 3 is explained. Furthermore, new design equations are presented by examining the intersection of the scan line with stability zone 3. Resolution versus transmission characteristics of stability zones 1 and 3 are compared and the dependence of performance for zones 1 and 3 is related to particular instrument operating parameters.


Brief strategic family therapy versus treatment as usual: results of a multisite randomized trial for substance using adolescents.
To determine the effectiveness of brief strategic family therapy (BSFT; an evidence-based family therapy) compared to treatment as usual (TAU) as provided in community-based adolescent outpatient drug abuse programs. A randomized effectiveness trial in the National Drug Abuse Treatment Clinical Trials Network compared BSFT to TAU with a multiethnic sample of adolescents (213 Hispanic, 148 White, and 110 Black) referred for drug abuse treatment at 8 community treatment agencies nationwide. Randomization encompassed both adolescents' families (n = 480) and the agency therapists (n = 49) who provided either TAU or BSFT services. The primary outcome was adolescent drug use, assessed monthly via adolescent self-report and urinalysis for up to 1 year post randomization. Secondary outcomes included treatment engagement (≥2 sessions), retention (≥8 sessions), and participants' reports of family functioning 4, 8, and 12 months following randomization. No overall differences between conditions were observed in the trajectories of self-reports of adolescent drug use. However, the median number of days of self-reported drug use was significantly higher, χ2(1) = 5.40, p < .02, in TAU (Mdn = 3.5, interquartile range [IQR] = 11) than BSFT (Mdn = 2, IQR = 9) at the final observation point. BSFT was significantly more effective than TAU in engaging, χ2(1) = 11.33, p < .001, and retaining, χ2(1) = 5.66, p < .02, family members in treatment and in improving parent reports of family functioning, χ2(2) = 9.10, p < .011. We discuss challenges in treatment implementation in community settings and provide recommendations for further research.


Analysis of cytokine secretion from lymphocytes of patients with hypersensitivity reactions to contaminated heparins.
Beginning in 2007, anaphylactoid reactions associated with unfractionated heparin (UFH) occurred and resulted in some fatalities. These reactions were reported to be linked to the complement and contact system activation induced by certain batches of UFH containing the adulterant oversulphated chondroitin sulphate (OSCS). Drug-specific secretion of selected cytokines from peripheral blood mononuclear cells (PBMC) of patients with hypersensitivity reactions to contaminated heparin was compared with the respective in vitro cytokine pattern of individuals with or without hypersensitivity to heparin, different glycosaminoglycans or other drugs. Study individuals (n = 13) were classified as follows: patients with hypersensitivity reactions to contaminated (OSCS) heparin (n = 3), noncontaminated heparin (n = 1) or other compounds (n = 3) and patients with ongoing heparin therapy without symptoms of intolerance (n = 2). Four healthy individuals served as controls. PBMC were incubated with six different glycosaminoglycan structures. Drug-specific intracellular interleukin (IL)-5, interferon (IFN)-γ and IL-10 production was investigated by flow cytometry, while secretion of IL-5, IL-2 and IFN-γ was analysed by enzyme-linked immunosorbent assay. PBMC from individuals with hypersensitivity reactions to contaminated heparin secreted considerable amounts of IL-2 in vitro. There was a suggestion that ongoing heparin therapy and the Li-heparin in the vials may have an impact on the lymphocyte reactivity of PBMC. The in vitro lymphocyte reactivity pattern of PBMC from individuals with hypersensitivity reactions to contaminated heparins was neither typical for an immune-mediated nor for a nonimmune-mediated reaction. Possible effects of heparins in the test system itself may require consideration.


Outcomes and complications of osseointegrated hearing aids in irradiated temporal bones.
To compare the complication rate for osseointegrated hearing aids (OIHA) in patients with or without irradiation. Retrospective case review. We studied patients with OIHAs implanted between January 1, 2005, and July 15, 2013 in a tertiary university center with a referral otology and neurotology practice. Demographics, history of oncologic surgery, follow-up length after OIHA implantation, radiation history and dosage, postoperative complications, and chronologic relationship between oncologic resection, OIHA implantation, and irradiation were reviewed to collect information. Soft tissue complications were graded according to a modified Holgers classification. The study included 48 patients. Twenty-nine patients (32 implants) did not undergo radiotherapy and 19 patients (19 implants) did. In the radiotherapy group, six patients had OIHAs implanted before radiotherapy, and 13 had OIHAs implanted in irradiated bone. Of these 13 patients, one had OIHA implanted during primary oncologic surgery; 11 had OIHA implanted during secondary surgery; and one patient did not have oncologic surgery. Patients with both OIHA implantation and radiotherapy had more complications than patients without radiotherapy (31.6% vs. 24.1%, P > 0.05) and more major complications than patients without radiotherapy (26.3% vs. 3.4%, P > 0.05). Patients with OIHAs implanted before radiotherapy did not have any complications. There were significantly fewer and less severe complications in patients with OIHAs implanted during primary oncologic resection than in patients with OIHAs implanted secondarily (0/8 vs. 8/11, P < 0.05). The rate and severity of complications of OIHAs can be minimized by implanting the device before irradiation, ideally at the time of primary oncologic surgery. 4. Laryngoscope, 126:1187-1192, 2016.


[Changes in the antidiuretic activity of the blood in hypertensive crises and experimental arterial hypertension of hypothalamic origin].
The blood antidiuretic activity was studied in patients with hypertensive disease in a period between crises and during a hypertensive crisis. The antidiuretic activity of blood was found to be higher in patients who had crises than in those with no history of crises. Blood antidiuretic activity diminishes during a hypertensive crisis, which is evidently due to the inhibiting effect on the hypothalamus of the receptors of the cardiovascular system. Similar results were obtained in experiments in elevation of arterial pressure induced by electric stimulation of the hypothalamic nuclei. It is concluded that the antidiuretic hormone plays a pathogenetic role in hypertensive disease which is marked by the development of crises.


Strategies to improve local control of resected pancreas adenocarcinoma.
Only approximately one in ten pancreas cancer patients is a candidate for potentially curative resection of this disease. Even this small fraction of patients has a poor prognosis following pancreatico-duodenectomy. The disease has an anatomic location that makes it difficult for the surgeon to maintain adequate margins of resection and prevent tumor spillage at the time of resection. Also, the disease is biologically aggressive and even with a complete visible resection of the disease, micrometastases are likely to remain behind. A survey of the sites for surgical treatment failure of resected pancreas cancer was performed. Also, the multiple modalities used in an attempt to improve the results of cancer resection are scrutinized. The surgical treatment failures are regional in nature and occur at the resection site and on peritoneal surfaces, within the liver, and within the regional lymph nodes. These anatomic sites account for nearly 100% of the initial sites of disease progression. Current hypothesis suggests that micrometastases released from the cancer specimen by the trauma of surgery account for the high incidence of resection site progression and peritoneal metastases. Although surgical trauma may contribute to micrometastases within the liver and lymph nodes, these are most likely present though not detected by preoperative radiologic studies. Adjuvant treatments such as neoadjuvant chemotherapy or combination systemic chemotherapy have not been associated with improved survival. Extended resections such as total pancreatectomy or extended lymphadenectomy have not been associated with benefit. However, resection with a negative margin of excision along with the removal of at least 12 lymph nodes in and around the pancreaticoduodenectomy specimen is associated with superior outcomes. A regional chemotherapy treatment that consists of hyperthermic intraperitoneal chemotherapy (HIPEC) with gemcitabine and long-term normothermic intraperitoneal chemotherapy (NIPEC-LT) gemcitabine for 6 months postoperatively is suggested as a new treatment that has demonstrated decreases in local-regional failure and promises to more adequately target micrometastases in the peritoneal space, in the liver and lymph nodes. Pancreas cancer surgery should attempt to achieve negative margins of resection with the removal of at least 12 lymph nodes. Hyperthermic intraperitoneal gemcitabine can adequately eradicate malignant cells dislodged from the cancer specimen into the bed of the resection at high density and on distant peritoneal surfaces as peritoneal metastases. Long-term intraperitoneal gemcitabine may act on micrometastases in the liver through absorption into the portal vein blood and the lymph nodes as a result of gemcitabine absorption by subperitoneal lymphatic channels. The use of HIPEC and NIPEC-LT gemcitabine may improve local control of resected pancreas cancer.


Intramyocardial Dissecting Hematoma: A Mechanical Complication Needing Surgical Therapy?
Intramyocardial dissecting hematoma is a form of cardiac rupture caused by myocardial infarction, percutaneous coronary intervention, or trauma. It is a cavity between myocardial fibers caused by partial rupture of the ventricular wall. Therapeutic management, including the timing for surgical approach, has not been standardized. We present a case series describing 4 patients. (Level of Difficulty: Intermediate.).


Major and trace element contents in apples from a cultivated area of central Greece.
Forty-two soil and apple samples from central Greece were collected and analyzed with regards to the content in major (Ca, Fe, K, Mg, Na, P, and S) and trace elements (As, B, Ba, Cd, Co, Cr, Cu, Hg, Mn, Mo, Ni, Pb, Sb, Sc, Se, Sr, Ti, and Zn). Soil samples were analyzed by inductively coupled plasma atomic emission spectrometry, while for the apples inductively coupled plasma mass spectrometry was implemented. Several elements such as As, B, Cd, Cr, Cu, Fe, Mn, Ni, Pb, Se, Ti, and Zn, represent high concentrations in apples from the study area. These relatively high concentrations may be a consequence of the local geology, along with the excessive application of agricultural products such as fertilizers and agrochemicals.


Enhanced killing of penicillin-treated S. aureus by host defences: effects of amoxycillin, cloxacillin and nafcillin in vitro and in experimental mastitis.
The effects of pretreatment of Staphylococcus aureus with subMIC or 10 x MIC amoxycillin, cloxacillin and nafcillin on subsequent killing by bovine neutrophils or sensitivity to lysostaphin were examined in vitro, using penicillinase positive and negative strains. All penicillins increased the susceptibility of S. aureus to bovine neutrophils; with amoxycillin this was most marked with the penicillinase negative strain. Cloxacillin pretreatment was significantly more effective than nafcillin or amoxycillin in some tests and equally effective in others. The sensitivity of S. aureus to lysostaphin was increased following cloxacillin or nafcillin exposure. In a mouse model of mastitis, more cloxacillin and nafcillin-treated S. aureus (at 1/4 MIC) were killed than untreated bacteria 1 hr after infection but by 4 hr no differences were found. No differences were seen after intramammary infection with 10 x MIC penicillin-treated S. aureus. In glands with established chronic mastitis both cloxacillin and nafcillin at high doses failed to kill staphylococci. These findings indicate that prior exposure to penicillins increases staphylococcal susceptibility to killing by neutrophils but following phagocytosis, intracellular bacteria are protected from the lethal action of penicillins.


Methods for ablating neurons.
This past year, laser ablation has been applied to investigations of neuromuscular connectivity in Drosophila, neuronal function in nematode, and mammalian central nervous system development. Ablation by targeted gene expression has been refined and applied to questions of neural development. Chromophore-assisted laser inactivation has been used to demonstrate distinct functions for two proteins during grasshopper neural development.


An OsCEBiP/OsCERK1-OsRacGEF1-OsRac1 module is an essential early component of chitin-induced rice immunity.
OsCEBiP, a chitin-binding protein, and OsCERK1, a receptor-like kinase, are plasma membrane (PM) proteins that form a receptor complex essential for fungal chitin-driven immune responses in rice. The signaling events immediately following chitin perception are unclear. Investigating the spatiotemporal regulation of the rice small GTPase OsRac1, we find that chitin induces rapid activation of OsRac1 at the PM. Searching for OsRac1 interactors, we identified OsRacGEF1 as a guanine nucleotide exchange factor for OsRac1. OsRacGEF1 interacts with OsCERK1 and is activated when its C-terminal S549 is phosphorylated by the cytoplasmic domain of OsCERK1 in response to chitin. Activated OsRacGEF1 is required for chitin-driven immune responses and resistance to rice blast fungus infection. Further, a protein complex including OsCERK1 and OsRacGEF1 is transported from the endoplasmic reticulum to the PM. Collectively, our results suggest that OsCEBiP, OsCERK1, OsRacGEF1, and OsRac1 function as key components of a "defensome" critically engaged early during chitin-induced immunity.


How do changes in the population tested for chlamydia over time affect observed trends in chlamydia positivity? Analysis of routinely collected data from young women tested for chlamydia in family planning clinics in the Pacific Northwest (USA), between 2003 and 2010.
Background The proportion of chlamydia tests that are positive (positivity) is dependent on the population tested and the test technology used. The way in which changes in these variables might affect trends in positivity over time is investigated. Data from 15- to 24-year-old women tested for chlamydia in family planning clinics participating in the Infertility Prevention Project in the Pacific Northwest, United States (USA Public Health Service Region X) during 2003-2010 (n=590557) were analysed. Trends in positivity and in test, demographic and sexual behaviour variables were identified. Unadjusted and adjusted trends in chlamydia positivity were calculated using logistic regression. The proportion of tests carried out using nucleic acid amplification tests (NAATs) increased dramatically during the analysis period in two states. Smaller changes in demographic and behavioural characteristics were seen. Controlling for test technology used had the largest effect on the trend in testing positive per year, leading to a fall in the calculated odds ratio of testing positive from 1.06 to 1.02 in Oregon, and from 1.07 to 1.02 in Idaho. Controlling for other variables had minimal effect on chlamydia positivity trends. Changes in NAAT use had a large effect on observed trends in chlamydia positivity over time in the two states where NAATs were introduced during the analysis period. While trends in chlamydia positivity may be a useful metric for monitoring chlamydia burden, it is important to consider changes in test type when interpreting these data.


GKLOMLI: a link prediction model for inferring miRNA-lncRNA interactions by using Gaussian kernel-based method on network profile and linear optimization algorithm.
The limited knowledge of miRNA-lncRNA interactions is considered as an obstruction of revealing the regulatory mechanism. Accumulating evidence on Human diseases indicates that the modulation of gene expression has a great relationship with the interactions between miRNAs and lncRNAs. However, such interaction validation via crosslinking-immunoprecipitation and high-throughput sequencing (CLIP-seq) experiments that inevitably costs too much money and time but with unsatisfactory results. Therefore, more and more computational prediction tools have been developed to offer many reliable candidates for a better design of further bio-experiments. In this work, we proposed a novel link prediction model based on Gaussian kernel-based method and linear optimization algorithm for inferring miRNA-lncRNA interactions (GKLOMLI). Given an observed miRNA-lncRNA interaction network, the Gaussian kernel-based method was employed to output two similarity matrixes of miRNAs and lncRNAs. Based on the integrated matrix combined with similarity matrixes and the observed interaction network, a linear optimization-based link prediction model was trained for inferring miRNA-lncRNA interactions. To evaluate the performance of our proposed method, k-fold cross-validation (CV) and leave-one-out CV were implemented, in which each CV experiment was carried out 100 times on a training set generated randomly. The high area under the curves (AUCs) at 0.8623 ± 0.0027 (2-fold CV), 0.9053 ± 0.0017 (5-fold CV), 0.9151 ± 0.0013 (10-fold CV), and 0.9236 (LOO-CV), illustrated the precision and reliability of our proposed method. GKLOMLI with high performance is anticipated to be used to reveal underlying interactions between miRNA and their target lncRNAs, and deciphers the potential mechanisms of the complex diseases.


Analysis of ictal EEGs of epilepsy associated with tuberous sclerosis.
To investigate the significance of cortical pathology of tonic spasms in patients with tuberous sclerosis. The subjects were 38 patients with epilepsy associated with tuberous sclerosis. We analyzed ictal EEGs of tonic spasms and partial seizures by means of video-EEG monitoring for a total of 763 tonic spasms in 20 patients and 107 partial seizures in 15 patients. We also investigated the relation between partial seizures and magnetic resonance imaging (MRI) findings of these patients. Ictal EEG patterns of tonic spasms were divided into generalized and focal patterns. Thirteen patients had only generalized patterns, whereas seven had both patterns. In five patients who had focal ictal patterns of tonic spasms and partial seizures, the location of focal patterns corresponded with the location of onset of partial seizures. Focal discharges were seen immediately before, after, and in the middle of tonic spasms in series in 13 patients. The location of focal discharges also corresponded with the location of the onset of partial seizures in 10 of the 13 patients. Regarding partial seizures, four patients had multiple active epileptogenic foci during the same period, and two others had shifting epileptogenic foci with increasing age. These findings indicate that cortical pathology plays an important role in the occurrence not only of partial seizures but also of tonic spasms in patients with tuberous sclerosis.


Comparative structural modeling and docking studies of uricase: possible implication in enzyme supplementation therapy for hyperuricemic disorders.
Uricase (EC 126.96.36.199, UC) catalyzes the oxidation of uric acid (UA) to more soluble allantoin thereby lowering plasma UA levels. In humans, when concentration of UA exceeds >7mg/dl, it leads to hyperuricemia, gout, nephrolithiasis and urolithiasis. A new remedy to cure such metabolic diseases is the enzyme supplementation therapy by UC but with high degree of antigenic independence. Therefore screening of new uricase sources to expand its usefulness and reduced antigenecity is needed. Present study employed cheminformatics approach to construct models of reported UC from different sources viz. Bacillus megaterium, Streptomyces bingchenggensis BCW-1, Paenibacillus sp, Solibacter usitatus Ellin6076, Truepera radiovictrix DSM 17093 and Ktedonobacter racemifer DSM 4496 in order to study their structure-function relationship for enzyme mass production and modification for improved characteristics. BioMed CAChe version 6.1 was further used to study enzyme-substrate interactions of models with uric acid using docking approach. Results indicated that models for UC of Streptomyces bingchenggensis BCW-1 accounted for better regio-specificity towards UA, supporting the interested metabolism and thus may further be implicated in enzyme supplementation therapy for hyperuricemic associated disorders.


Net growth delay: a novel parameter derived from tumor growth curves.
Growth delay does not only reflect the effect of treatment on the tumor parenchymal cells but also on the stroma. Due to tumor bed effect, the extent of growth delay determined from tumor growth curves is highly dependent on the end volume chosen. It was aimed to minimize the influence of the tumor bed effect on the growth delay calculated by choosing a smaller size and essentially an earlier time for regrowth. Net growth delay is a novel parameter derived from the tumor growth curves, allowing a better comparison of the results with colony assay and tumor control data.


[The effect of ultrasonic oscillations and amphotericin B on the ultrastructure of yeast-like fungi in the genus Candida].
C. albicans cellular structure was examined electron microscopically over the course of amphotericin B treatment. The efficacy of amphotericin B administration via phonophoresis is demonstrated.


Silicone-specific imaging using an inversion-recovery-prepared fast three-point Dixon technique.
To demonstrate a new hybrid magnetic resonance imaging (MRI) technique capable of simultaneously generating water-specific and silicone-specific images in a single acquisition. This technique combines short TI inversion-recovery (STIR) technique for robust fat suppression with an efficient fast spin-echo-based three-point Dixon technique for robust separation of remaining water and silicone in the presence of field inhomogeneities. Images demonstrating the feasibility of the technique were acquired with a 1.5-Tesla scanner in a phantom and in a volunteer with both saline and silicone implants in vivo. The new technique provided water-specific and silicone-specific images of diagnostic quality. Separation of the water and silicone chemical species was complete and satisfactory. Compared with a chemical shift-selective technique, the new technique does not rely heavily on field homogeneity and requires the same or even less scan time to acquire images with similar scan parameters, resolution, and signal-to-noise ratio (SNR). The feasibility and potential application of the new technique were demonstrated via imaging a phantom and a silicone breast prosthesis in vivo, and it may be used for more consistent imaging of the silicone implants without compromising the image quality or overall scan time.


Midline crossing pulmonary vein: right upper lobe dual venous drainage, with partial anomalous venous return of the right lung into a persistent left superior vena cava.
We present a case of dual drainage of the right upper lobe of the lung into the left atrium and via partial anomalous venous pulmonary return (PAPVR) into a persistent left superior vena cava (SVC). It is only in the minority of PAPVR cases where the anomalous pulmonary veins cross the midline. We provide a review of current literature on this topic and an explanatory embryological model. Knowledge of embryonic development and possible anatomic variations, including the concept of dual venous drainage of the lung, leads to better interpretation of imaging, with more accurate description of the morphology at hand. High-resolution multidetector computed tomography (MDCT) helps to delineate the exact vascular anatomy. This will enhance a better understanding of and anticipation on the patient's disease status, with more accurate planning of intervention, and possibly less complications.


Development, feasibility, and acceptability of an intervention to improve care for agitation in people living in nursing homes with dementia nearing the end-of-life.
To develop a staff training intervention for agitation in people with severe dementia, reaching end-of-life, residing in nursing homes (NHs), test feasibility, acceptability, and whether a trial is warranted. Feasibility study with pre- and post-intervention data collection, qualitative interviews, and focus groups. Three NHs in South East England with dementia units, diverse in terms of size, ownership status, and location. Residents with a dementia diagnosis or scoring ≥2 on the Noticeable Problems Checklist, rated as "severe" on Clinical Dementia Rating Scale, family carers, and staff (healthcare assistants and nurses). Manualized training, delivered by nonclinical psychology graduates focusing on agitation in severe dementia, underpinned by a palliative care framework. Main outcomes were feasibility of recruitment, data collection, follow-up, and intervention acceptability. We collected resident, family carer, and staff demographics. Staff provided data on resident's agitation, pain, quality of life, and service receipt. Staff reported their sense of competence in dementia care. Family carers reported on satisfaction with end-of-life care. In qualitative interviews, we explored staff and family carers' views on the intervention. The target three NHs participated: 28 (49%) residents, 53 (74%) staff, and 11 (85%) family carers who were eligible to participate consented. Eight-four percent of staff attended ≥3 sessions, and we achieved 93% follow-up. We were able to complete quantitative interviews. Staff and family carers reported the intervention and delivery were acceptable and helpful. The intervention was feasible and acceptable indicating a larger trial for effectiveness may be warranted.


A 3-Day residential yoga-based program improves education professionals' psychological and occupational health in a single arm trial.
This study examined changes in psychological and occupational health in urban education professionals after attending a brief yoga-based program. Education professionals from the New York City Department of Education (NYC DOE) who were attending a residential 3-day yoga-based program at Kripalu Center for Yoga & Health were recruited to participate in the study. Measures of psychological and occupational health and health-related behaviors were completed before (baseline), after (post), and two months after the program (follow-up). Paired samples t-tests were used to compare scores between time points. At post, participants (N = 74) showed improvements in stress, resilience, affect, mindfulness, empowerment, self-compassion, satisfaction with life, work engagement, burnout, exercise, and vegetable intake (all p values < 0.05) compared to baseline. At the follow-up (N = 33), showed improvements in resilience, affect, mindfulness, empowerment, self-compassion, work engagement, and burnout (all p values < 0.05) compared to baseline. There were significant correlations between the degree of home practice of the skills and techniques learned in the program and improvements in multiple measures of psychological and occupational health at follow-up (all p values < 0.05). These findings suggest that the yoga-based program improves psychological and occupational health and healthy behaviors in education professionals immediately following the program and up to two-months following the program, however, more data with larger sample sizes are needed to confirm sustained benefits over the longer term.


Flea species from dogs and cats in northern Greece: environmental and clinical implications.
Fleas were identified after being collected from 129 dogs and 38 cats of random breed, sex and age. All these animals, infested with fleas and admitted to the Clinic of Medicine of the Veterinary Faculty in Thessaloniki for routine procedures, were from different habitats and originated from various parts of northern Greece. Ctenocephalides canis was the most common species found on the dogs (71.3%). Conversely, its prevalence in the 14 cats was substantially lower (5.3%). Ctenocephalides felis was found on 97.4% of the cats and 40.3% of the dogs surveyed. Of the other flea species, with much lower prevalence, Pulex irritans (0.8%) and Xenopsylla cheopis (0.8%) were observed only on the dogs. Seventeen dogs (13.2%) and one cat (2.6%) had mixed infestations. Flea-associated dermatoses were observed in 26 dogs (20.2%) and four cats (10.5%). Flea-allergic dermatitis, with its typical manifestations, was seen in ten of the dogs (38.5%) with skin lesions. Three out of four flea-allergic cats presented miliary dermatitis and one symmetrical hypotrichosis.


Applying the 4Ps of social marketing to retain and engage participants in longitudinal cohort studies: generation 2 Raine study participant perspectives.
Investigations of participant retention in longitudinal health and medical research, document strategies that work best but overlook social marketing's capacity to influence participant retention. After applying the social marketing framework: the idea that determining what longitudinal participants 'buy' (product), at what cost (price), in what location (place) and through which communication channels (promotion), this paper aims to inform and enhance retention efforts. This qualitative study was conducted through in-depth interviews with participants from the Raine Study that began in Western Australia in 1989. The Generation 2 participants, initially enrolled into the Raine Study as babies by their parents (Generation 1), are now young adults invited to attend follow-up studies and tests every few years. Our study defined 'active' participants (n = 17) as those who agreed to attend their 27 year follow-up, and 'inactive' (n = 12) participants as those who had attended neither of the past two follow-ups (22 and 27 years). Raine Study participants experienced core, actual and augmented product benefits. Inactive participants focused on the costs (price) associated with participation, and were more likely to suggest tele-health (place) strategies to overcome barriers to follow-up attendance. Both active and inactive participants found professional processes and friendly staff made the Raine Study environment appealing, suggested that social media (promotion) was underutilised, and offered novel ideas to enhance engagement. Social marketing can support the development of differentiated strategies addressing the unique needs and wants of active and inactive participants. Sophisticated cohort segmentation can reach participants in a more meaningful way, reinforce the study 'brand' and guard against attrition.


On the relationship between deterministic and probabilistic directed Graphical models: from Bayesian networks to recursive neural networks.
Machine learning methods that can handle variable-size structured data such as sequences and graphs include Bayesian networks (BNs) and Recursive Neural Networks (RNNs). In both classes of models, the data is modeled using a set of observed and hidden variables associated with the nodes of a directed acyclic graph. In BNs, the conditional relationships between parent and child variables are probabilistic, whereas in RNNs they are deterministic and parameterized by neural networks. Here, we study the formal relationship between both classes of models and show that when the source nodes variables are observed, RNNs can be viewed as limits, both in distribution and probability, of BNs with local conditional distributions that have vanishing covariance matrices and converge to delta functions. Conditions for uniform convergence are also given together with an analysis of the behavior and exactness of Belief Propagation (BP) in 'deterministic' BNs. Implications for the design of mixed architectures and the corresponding inference algorithms are briefly discussed.


Elemental mapping of Moroccan enameled terracotta tile works (Zellij) based on X-ray micro-analyses.
The purpose of this work is the elemental mapping of enameled terracotta samples (Zellij), produced between the 13th and 20th centuries in Morocco, collected from five different monuments from Marrakech. These pieces were analyzed by two non-destructive micro X-Ray Fluorescence (XRF) spectrometers, aiming to obtain elemental distribution and elemental composition. From the obtained spectra we have identified the main elements present in the tin-opacified lead glaze. The identification of the decoration colors is based on the different ratios between the fluorescence lines of the main component of the glaze (Pb-Lα line) and the fluorescence lines of the main components of the pigment (Co-Kα, Mn-Kα, Ni-Kα,… lines). The semi-quantitative calculations based on these ratios revealed significant differences between modern and ancient samples.


[Clinical aspects and radiology of histiocytosis X in the skeleton--a retrospective study of 18 patients].
The present paper presents the clinical and radiological findings from a retrospective study of 18 children suffering from histiocytosis with skeletal involvement (20 lesions) treated between 1971 and 1983. The different forms of manifestation on the skull, vertebral column, and long bones are described and problems of differential diagnosis discussed. Possible forms of therapy are considered.


Teledentistry at the Crossroads: Benefits, Barriers, and Beginnings.
Since the start of COVID-19 shelter-in-place orders given as early as March 16, 2020, routine dental care has experienced a sharp interruption. This challenging time has provided an opportunity for healthcare professionals to use and collaborate with available technologies in a meaningful way. One of the silver linings for dental professionals during this time has been the use of teledentistry. While it is not a replacement for a clinical examination, studies show that teledentistry can be utilized to triage cases, maintain contact with patients, and successfully identify abnormal oral lesions. This article explains both the benefits and barriers of teledentistry that specifically apply to pandemic conditions, providing examples of how this modality can be used by different specialties in dentistry. Additionally, the article proposes models of how teledentistry can be effectively integrated in two different settings: the private practice model and dental schools.


What makes a difference: a method to determine whether a change in an image affects the perceived gist.
Although many changes in images and real-world situations can go undetected, researchers have suggested that changes affecting the gist of an image are likely to be noticed. This article proposes a method to determine whether a change in an image affects its gist. The method was designed to determine a systematical difference between sets of descriptions of subjectively interpretable images. First, for each image, a set of descriptions is generated. Then, a rater judges good fit or poor fit for the descriptions for each image, providing the base for categorization. Finally, a Fisher exact probability test is used to determine a difference between sets of descriptions. The method is applied to determine a gist change for 18 test sets. The proposed method makes it possible to compare subjective interpretations of images, while leaving room for differences in interpretation of an image by different people.


Comparison of dopants for charge exchange ionization of nonpolar polycyclic aromatic hydrocarbons with reversed-phase LC-APPI-MS.
Atmospheric pressure photoionization (APPI) is capable of ionizing nonpolar compounds in LC/MS, through charge exchange reactions following photoionization of a dopant. Recently, several novel dopants-chlorobenzene, bromobenzene, 2,4-difluoroanisole, and 3-(trifluoromethyl)anisole-have been identified as having properties making them well-suited to serve as dopants for charge exchange ionization under reversed-phase LC conditions. Here, we report the results of experiments comparing their effectiveness to that of established dopants-toluene, anisole, and a toluene/anisole mixture, for the charge exchange ionization of model nonpolar compounds-the 16 polycyclic aromatic hydrocarbons (PAHs) identified by the US EPA as priority pollutants-when using a conventional reversed-phase LC method. Chloro- and bromobenzene were found to be much more effective than toluene for all the PAHs, due to the relatively low reactivity of their photoions with the solvent. Their overall performance was also better than that of anisole, due to anisole's ineffectiveness toward higher-IE compounds. Further, the experiments revealed that anisole's performance for higher-IE compounds can be dramatically improved by introducing it as a dilute solution in toluene, rather than neat. The two fluoroanisoles provided the highest overall sensitivity, by a slim margin, when introduced as dilute solutions in either chloro- or bromobenzene.


Heterogeneity of associations between dissociation and attention deficit symptoms.
Childhood trauma and dissociative experiences are suggested to be predisposing transdiagnostic factors for attention deficit /hyperactivity disorder (ADHD) as well as many psychiatric disorders. Trauma-related symptoms such as motor restlessness, emotional instability, and concentration problems can mimic, trigger, or exacerbate ADHD symptoms. Moreover, given the relationship between ADHD and trauma-induced distress, it has been suggested that dissociative experiences and attention problems may reveal overlapping characteristics. The aim of this study was to investigate the associations between dissociative experiences and attention deficits by carrying out mixture analysis. A thousand and thirty-seven participants volunteered to the online investigation. Participants completed a test battery that included a sociodemographic form as well as the Adult ADHD Severity Rating Scale (ASRS), Wender Utah Rating Scale (WURS), Childhood Trauma Questionnaire (CTQ), Dissociative Experiences Scale (DES), and Somatoform Dissociation Questionnaire (SDQ). Item responses on the DES and attention deficit symptoms as indexed by the ASRS were subjected to latent class analysis. The three-latent-class model outperformed alternative mixture models. Mixture analysis classified the sample into three homogenous subgroups as follows: (1) No/low dissociation or attention problems; (2) Moderate dissociation with attention problems; and (3) High dissociation with attention problems. High dissociators with attention problems were characterized by heightened scores on somatoform dissociation and emotional neglect. No/low dissociation or attention problems latent class reported significantly lower scores on hyperactivity/impulsivity, depression, attention deficit in childhood, and sexual abuse than both moderate and high dissociation latent classes. High dissociators and moderate dissociators significantly differed on conduct problems and physical abuse. We concluded that impulsive hyperactivity, depression, and childhood sexual abuse were common features in heightened dissociation latent classes, as well as attention deficit.


Screening hospital admissions from the emergency department for occult carbon monoxide poisoning.
Because cases of unrecognized carbon monoxide (CO) poisoning have been described among patients admitted to the hospital with other diagnoses, screening hospital admissions with carboxyhemoglobin testing has the potential for preventing morbidity among patients as well as among their cohabitants. Carboxyhemoglobin levels were obtained on 753 patients admitted to the hospital from the emergency department over a 3-month period during the winter. Patients in whom CO poisoning was diagnosed in the emergency department prior to admission were excluded. The mean carboxyhemoglobin level was 2.52% +/- 1.85%; there was no significant difference in mean carboxyhemoglobin among patients with medical, surgical, neurological, and psychiatric admission diagnoses (F = 1.17; df = 3,746; P = .32). Two patients (0.3%; 95% confidence limits, 0.04% to 1.1%) from the entire admission cohort, and one of 20 patients (5%; 95% confidence limits, 0.3% to 26.9%) admitted with seizures, had carboxyhemoglobin levels greater than 10%. The carboxyhemoglobin levels of the two patients were only marginally elevated, with levels of 10.9% and 11.3%. The cost of the carboxyhemoglobin screening program was $2.26 per patient result, or approximately $2,100 over a 3-month winter heating season. A program for screening emergency department admissions with carboxyhemoglobin testing, although feasible in terms of cost, detected few cases of unrecognized CO poisoning.


Reproductive aging results in a reconfigured ovarian antioxidant defense profile in rats.
To test our hypothesis that reproductive aging changes the ovarian oxidative stress defense profile, in response to prostaglandin F2alpha (PGF2alpha) during corpus luteum regression, because how a cell or an organ handles reactive oxygen intermediates may be dependent on the biological age of the organism. Animal experimentation using rat model of corpus luteum regression. University reproductive biology laboratory. Control (26-day-old) and 8- to 9-month-old (reproductive aging) rats. Corpus luteum formation was induced in control and 8- to 9-month-old (reproductive aging) rats with pregnant mare serum gonadotropin followed by human chorionic gonadotropin. Regression was then initiated with PGF2alpha. Vitamin E, glutathione reductase, glutathione peroxidase, catalase, and thiobarbituric acid-reacting substances were measured. Ovaries from reproductive aging rats, compared with the control (26-day-old) group, had elevated vitamin E levels at 0, 2, and 24 hours after PGF2alpha. At 2 and 24 hours after PGF2alpha, the aging ovaries had lower glutathione reductase levels. These data suggest that the reproductive aging ovary has a transformed oxidative stress defense profile and that this may account for some of the physiological changes found in reproductive aging.


Ultraviolet light affects the color vocabulary: evidence from 834 languages.
It has been suggested that people living in regions with a high incidence of ultraviolet light, particularly in the B band (UV-B), suffer a phototoxic effect during their lifetime. This effect, known as lens brunescence, negatively impacts the perception of visible light in the "blue" part of the spectrum, which, in turn, reduces the probability that the lexicon of languages spoken in such regions contains a word specifically denoting "blue." This hypothesis has been recently tested using a database of 142 unique populations/languages using advanced statistical methods, finding strong support. Here, this database is extended to 834 unique populations/languages in many more language families (155 vs. 32) and with a much better geographical spread, ensuring a much better representativity of the present-day linguistic diversity. Applying similar statistical methods, supplemented with novel piecewise and latent variable Structural Equation Models and phylogenetic methods made possible by the much denser sampling of large language families, found strong support for the original hypothesis, namely that there is a negative linear effect of UV-B incidence on the probability that a language has a specific word for "blue." Such extensions are essential steps in the scientific process and, in this particular case, help increase our confidence in the proposal that the environment (here, UV-B incidence) affects language (here, the color lexicon) through its individual-level physiological effects (lifetime exposure and lens brunescence) amplified by the repeated use and transmission of language across generations.


Prefilled syringe needles versus standard removable needles for intravitreous injection.
To investigate differences in the scleral resistance between standard disposable 27 gauge (G) needle versus the Macugen prefilled syringe needle. Observational study at one eye center. One injecting physician performed 46 intravitreous injections using standard Becton Dickinson 27 G needles and 71 intravitreous injections using Macugen needles during January to May 2005. The procedure in two patients was videotaped and analyzed frame by frame. Microphotographs of used needles were analyzed. There were no complications during the injection procedure in either group. Microphotographic analysis after injection revealed 3 and 5 facets on standard 27 G needle and Macugen needles, respectively. The bevel forming the tip of the 27 G needle was steeper and longer than that of the Macugen needle. The inside diameter in the standard 27 G needle was 220 microm versus 210 microm in the Macugen needle. There are differences in needle design between commonly used disposable 27 G needles and 27 G Macugen needles which result in higher resistance to penetration with Macugen needle. Careful needle design is an important concern in manufacturing prefilled syringes with needles attached to them.


Catalytic Asymmetric Direct Vinylogous Aldol Reaction of Isatins with β,γ-Unsaturated Butenolides.
A direct asymmetric vinylogous aldol reaction of nonactivated natural α-angelica lactone to isatins has been developed. With a N,N'-dioxide-Sc(OTf)3 complex as catalyst, a variety of δ-hydroxy butenolides bearing congested adjacent tetrasubstituted stereocenters were obtained in good yields with high diastereoselectivities and excellent enantioselectivities. Besides, a possible transition state was proposed to explain the origin of the asymmetric induction.


Spinal cord long-term potentiation is attenuated by the NMDA-2B receptor antagonist Ro 25-6981.
The NR2B-containing N-methyl-d-aspartate (NMDA) receptors may be involved in a variety of phenomena including synaptic plasticity, memory formation and pain perception. Here we used the NMDA-2B receptor antagonist Ro 25-6981 to investigate the role of the NR2B-containing NMDA receptors in spinal nociception. Extracellular single unit recordings were performed from dorsal horn wide dynamic range (WDR) neurones in intact urethane-anaesthetized Sprague-Dawley rats. The responses of the WDR neurones evoked by C-fibre activation after sciatic nerve stimulation were defined according to latencies. To block the dorsal horn NMDA-2B receptors, the antagonist Ro 25-6981 was applied topically onto the spinal cord. High-frequency stimulation (HFS) of the sciatic nerve was used to induce spinal long-term potentiation (LTP). Spinal administration of the NMDA-2B receptor antagonist Ro 25-6981 had a clear antinociceptive effect at the spinal level (P < 0.05, C-fibre evoked responses after 4 mm Ro 25-6981 vs. C-fibre evoked responses in baseline). Moreover, spinal administration of this antagonist clearly attenuated the magnitude of spinal cord LTP after HFS conditioning (P < 0.05, C-fibre evoked responses after HFS vs. C-fibre evoked responses after 8 mm Ro 25-6981 + HFS). The present study indicates that expression of full LTP in dorsal horn neurones obtained by HFS conditioning may be dependent on the NMDA receptors containing the NR2B subunit. This suggests that activation of dorsal horn NR2B-containing NMDA receptors may be involved in use-dependent sensitization at the spinal level.


Finite element analysis regarding patch size, stiffness, and contact condition to the endocardium in surgery for post infarction ventricular septal rupture.
The purpose of this study is to establish a rational surgical design to minimize suture line stress of the patch and thus prevent residual leakage in surgery for post infarction ventricular septal rupture (VSR). A computer model that simulates surgery for VSR was developed. Stress force on the suture line of the patch was calculated at varying size, stiffness, and contact condition of the endocardial patch to the inner surface of the heart using a finite element analysis. Clinical results and echo findings of 34 consecutive patients with a mean age of 72.6 ± 9.5 (range 55-89) who underwent emergency surgery for VSR from 1995 to 2012 were reviewed. Suture line stress decreased by two-thirds as the size or stiffness of the patch increased in comparison with the basic conditions that mimic a pericardial patch fitted to the septal plane. On the other hand, suture line stress increased sixfold when there was a dead space beneath the patch. 30-day mortality was 12 %, and in-hospital mortality 18 %. On echocardiography, all three patients who had dead space beneath the patch had residual leak. Another patient who had huge posterior defect also showed residual leak. Clinical results were well matched to model results. 5-year survival rate of all patients who received operation was 68.7 ± 9.3 %. In endocardial patch type surgery for VSR, proper sizing of the patch to sufficiently fit to endocardial surface in a tension-free condition is the most important to avoid residual leak.


Early increase in alveolar macrophage prostaglandin 15d-PGJ2 precedes neutrophil recruitment into lungs of cytokine-insufflated rats.
Early detection and prevention is an important goal in acute respiratory distress syndrome research. We determined the concentration of the anti-inflammatory 15-deoxy-Δ(12,14)-prostaglandin-J2 (15d-PGJ2) and other components of the cyclopentenone prostaglandin cascade in relation to lung inflammation in cytokine (IL-1/LPS)-insufflated rats. We found that 15d-PGJ2 levels increase in the bronchoalveolar lavage (BAL) fluid of rats insufflated with cytokines 2 h before. BAL 15d-PGJ2 increases preceded neutrophil recruitment, lung injury, and oxidative stress in the lungs of cytokine-insufflated rats. 15d-PGJ2 was localized in alveolar macrophages that decreased following cytokine insufflation. 15d-PGJ2 may constitute an early biomarker of lung inflammation and may reflect an endogenous attempt to regulate ongoing inflammation in macrophages and elsewhere after cytokine insufflation.


Association between angiotensin-converting enzyme inhibitors and the risk of lung cancer: a systematic review and meta-analysis.
The association between the use of angiotensin-converting enzyme inhibitors (ACEIs) and lung cancer risk remains controversial. This study evaluated the association between the use of ACEIs and lung cancer risk. Records from five databases were searched from inception to 26 January 2022. Clinical studies involving persons aged ≥18 years with at least one year of follow-up and reporting adverse events, including lung cancer, were recorded with separate outcome reports supplied for the ACEIs and control groups. Data were extracted independently by three authors and pooled using a random-effects model. The primary outcome was lung cancer development. Odds ratios (ORs) with 95% confidence intervals (CIs) and lung cancer-related morbidity were calculated. Of 2400 records screened, 13,061,226 patients were included from seven cohort studies and four case-control studies. Pooled results showed that ACEIs use was linked to increased lung cancer risk (OR 1.19, 95% CI 1.05-1.36; P = 0.008), with high heterogeneity (I2 = 98%). ACEI usage is a greater risk factor for lung carcinogenesis than angiotensin receptor blocker use, especially in Asian patients. Further randomised controlled trials are needed to confirm the causal association between the use of ACEIs and lung cancer risk.


Proteomic dataset of the organohalide-respiring bacterium Dehalococcoides mccartyi strain CBDB1 grown on hexachlorobenzene as electron acceptor.
The proteome of the anaerobic organohalide-respiring bacterium Dehalococcoides mccartyi strain CBDB1 was analyzed by nano liquid chromatography coupled to mass spectrometry (LC-MS/MS). Two different preparation methods, (i) in-solution and (ii) in-gel proteolytic digestion were assessed to elucidate the core and the functional proteome of bacterial cultures grown in synthetic anaerobic medium with hexachlorobenzene as sole electron acceptor. A detailed analysis of the data presented is available (Schiffmann et al., 2014) [1].


Preparation of novel clay/chitosan/ZnO bio-composite as an efficient adsorbent for tannery wastewater treatment.
This study focuses on the preparation of an activated clay/chitosan/ZnO bio-composite using solvent casting method. Clay was activated through microwave radiation using 1 M H2SO4 at a minimum liquid to solid ratio (L/S). Chitosan was extracted from waste prawn shell and ZnO nanoparticles (ZnO-NPs) were synthesized from zinc acetate di-hydrate (Zn (CH3CO2)2·2H2O) using the sol-gel method. The produced bio-composite were characterized using FT-IR, TGA, XRD and SEM. Response surface methodology (RSM) was used for experimental design to find out the optimum conditions, e.g., pH of the solution, dosage of adsorbent and contact time for the removal of methylene blue (MB) and Cr (VI) using MINITAB 18.1 software. The optimum conditions obtained for the highest removal of MB were pH 9.57, dosage 55.44 mg and contact time 114.09 min. Similarly, for the highest removal of Cr (VI) the optimum conditions were pH 3.75, dosage 67.42 mg and contact time 111.27 min. Applying these optimum conditions, the highest removal efficiency for MB and Cr (VI) was obtained at 84.21 % and 82.67 % with 9.57 mg g-1 and 10.45 mg g-1 of adsorption capacity respectively. The adsorption data were studied for both Langmuir and Freundlich isotherm. The value of maximum Langmuir sorption was (qm) 17.346 mg g-1 and 17.621 mg g-1 for MB and Cr (VI) respectively.


Beclomethasone dipropionate and formoterol reduce oxidative/nitrosative stress generated by cigarette smoke extracts and IL-17A in human bronchial epithelial cells.
Interleukin-17A (IL-17A), cigarette smoke and oxidative/nitrosative stress are involved in inflammatory airway diseases, and the mechanisms behind these processes are still poorly understood. We investigated whether recombinant human IL-17A (rhIL-17A), in combination with cigarette smoke extracts (CSE), increases the levels of inducibile nitric oxide synthase (iNOS), reactive oxygen species, nitrotyrosine (NT) and the activation of signal transducer and activator of transcription 1 (STAT-1) in normal human bronchial epithelial cells (16HBE). The effect of beclomethasone dipropionate (BDP), formoterol and their combination was also evaluated. We demonstrated that rhIL-17A or CSE alone increases iNOS expression, reactive oxygen species and NT production and STAT-1 downstream signalling activation in terms of STAT-1ser727 and STAT-1tyr701 phosphorylation. The combination of both stimuli further increased iNOS, ROS, NT and STAT-1ser727 phosphorylation. The silencing of STAT-1 expression partially reduced the levels of iNOS, reactive oxygen species and NT generated by rhIL-17A and inhibited the effect of CSE alone in 16HBE cells. The treatment of the cells with the MEK1/2 inhibitor U0126 (1,4-diamino-2,3-dicyano-1,4-bis (o-aminophenylmercapto butadiene) abolished the expression of iNOS and STAT-1ser727 phosphorylation generated by rhIL-17A. 16HBE treated with BDP or formoterol alone partially suppressed the effect of IL-17A or CSE on ROS, NT, and STAT-1 activation. Furthermore the use of the drugs in combination showed an additive effect in 16HBE. Our findings demonstrate that IL-17A increases oxidative/nitrosative markers, likely via ERK1/2 downstream signalling and STAT-1 pathway activation in human bronchial epithelial cells. BDP and formoterol treatment reduces this effect showing an additive effect used in combination.


Fourier transform coupled to tryptophan-scanning mutagenesis: lessons from its application to the prediction of secondary structure in the acetylcholine receptor lipid-exposed transmembrane domains.
Although Fourier transform (FT) and tryptophan-scanning mutagenesis (TrpScanM) have been extremely useful for predicting secondary structures of membrane proteins, they are deemed to be low-resolution techniques. Herein, we describe the combined use of FT and TrpScanM (FT-TrpScanM) as a more reliable approach for the prediction of secondary structure. Five TrpScanM studies of the acetylcholine receptor lipid-exposed transmembrane domains (LETMDs) were revisited and analyzed by FT-TrpScanM. FT analysis of the raw data from the aforementioned TrpScanM studies supports and validates the conclusions derived from their tryptophan-periodicity profiles. Furthermore, by FT-TrpScanM, we were able to determine the minimum number of consecutive tryptophan substitutions necessary for more robust prediction of alpha-helical secondary structures and evaluate the quality of structure predictions by alpha-helical character curves. Finally, this study encourages future utilization of FT-TrpScanM to more reliably predict secondary structures of the membrane protein LETMDs.


Characterization of monoclonal antibodies specific for isopentenyl adenosine derivatives occurring in transfer RNA.
A family of isopentenyl adenosine derivatives are naturally occurring components of transfer RNA and are involved in several different functional roles in the cell. To facilitate the study of the biochemistry of these modified nucleosides we have raised monoclonal antibodies to N6-(delta 2-isopentenyl)adenosine and N6-(4-hydroxy-3-methyl-but-2-enyl)adenosine. The antibodies show considerable specificity and three characteristic types are distinguishable. The first type have the hydroxylated derivative as the preferred antigen, the second type have isopentenyl adenosine as the preferred antigen and a third type show a specificity for all isopentenyl-containing derivatives.


Nuclear magnetic resonance-based metabolomics reveals similar metabolomics profiles in undifferentiated peripheral spondyloarthritis and reactive arthritis.
After exclusion of reactive, psoriatic and enteritis-associated arthritis, a group of "undifferentiated" peripheral spondyloarthritis (pSpA) remains. This group shares genetics, T-cell repertoire, and cytokines with reactive arthritis (ReA). ReA is preceded by gut or urogenital infection. Otherwise the two may be similar. We know little of the metabolic pathways driving undifferentiated pSpA or ReA. Nuclear magnetic resonance (NMR)-based metabolomics presents a hypothesis-free approach to study and compare metabolic pathways driving undifferentiated pSpA and ReA. Serum and synovial fluid metabolomes of 19 ReA and 13 undifferentiated pSpA, and serum metabolome of 18 controls were profiled using 1 H-based NMR. Partial least square-discriminant analysis (PLS-DA) identified metabolites different in patients as compared to controls. Multivariate analysis confirmed these. Altered metabolic pathways were identified using metabolites set enrichment analysis (MSEA). The serum and synovial fluid metabolomes of ReA and undifferentiated pSpA were compared. Ten serum metabolites of ReA/undifferentiated pSpA were different from those of controls. Six metabolites were different between serum and synovial fluid of these patients. MSEA identified five pathways different between patients and controls, and five pathways different between serum and synovial fluid of patients. PLS-DA showed no difference between the metabolomes of serum or of synovial fluid between ReA and undifferentiated pSpA. Identified metabolic pathways may be explored further to understand the pathogenesis and to target therapeutics. The similar immuno-metabolic pathways suggest similar pathogenesis of ReA and undifferentiated pSpA. Thus, they should be studied as a single disease entity.


Bioremediation of reject water from anaerobically digested waste water sludge with macroalgae (Ulva lactuca, Chlorophyta).
Phosphorus and biologically active nitrogen are valuable nutrient resources. Bioremediation with macroalgae is a potential means for recovering nutrients from waste streams. In this study, reject water from anaerobically digested sewage sludge was successfully tested as nutrient source for cultivation of the green macroalgae Ulva lactuca. Maximal growth rates of 54.57±2.16% FW d(-1) were achieved at reject water concentrations equivalent to 50 μM NH4(+). Based on the results, the growth and nutrient removal was parameterised as function of NH4(+) concentration a tool for optimisation of any similar phycoremediation system. Maximal nutrient removal rates of 22.7 mg N g DW(-1) d(-1) and 2.7 mg P g DW(-1) d(-1) were achieved at reject water concentrations equivalent to 80 and 89 μM NH4(+), respectively. A combined and integrated use of the produced biomass in a biorefinery is thought to improve the feasibility of using Ulva for bioremediation of reject water.


Analytical approach to time lag in binary nucleation.
We present an analytical formula for the time required to establish steady state in a nucleating binary system. To test our solution, we evaluate the time lag for a range of activities of both components at the vapor-liquid transition, and show that our result is in much better agreement with a purely numerical simulation than other available analytical formulas, which overestimate the time lag by factors of from 2 to 200.


Slow alpha variant during REM sleep.
REM sleep resembles wakefulness or drowsiness. The pattern can be low-voltage, or else alpha-like. Alpha frequencies are present only during the tonic phases of REM sleep and, compared to wakefulness tracings, alpha activities are slightly slower by 1-2 Hz and are more monomorphic. The slow alpha-variant rhythm or subharmonic alpha pattern consists of rhythmic notched theta waves. It shares the same topography and reactivity with the alpha rhythm. We report its presence during the tonic phases of REM sleep without any modification in its morphology in the first case. In the second case, its morphology was similar to awakening but with slower amplitude. No alpha frequencies were found in REM sleep but only its slow alpha variant. This study provides evidence that REM sleep is a stage of sleep that contains rhythms similar to those seen during wakefulness and drowsiness.


Limits of variation, specific infectivity, and genome packaging of massively recoded poliovirus genomes.
Computer design and chemical synthesis generated viable variants of poliovirus type 1 (PV1), whose ORF (6,189 nucleotides) carried up to 1,297 "Max" mutations (excess of overrepresented synonymous codon pairs) or up to 2,104 "SD" mutations (randomly scrambled synonymous codons). "Min" variants (excess of underrepresented synonymous codon pairs) are nonviable except for P2Min, a variant temperature-sensitive at 33 and 39.5 °C. Compared with WT PV1, P2Min displayed a vastly reduced specific infectivity (si) (WT, 1 PFU/118 particles vs. P2Min, 1 PFU/35,000 particles), a phenotype that will be discussed broadly. Si of haploid PV presents cellular infectivity of a single genotype. We performed a comprehensive analysis of sequence and structures of the PV genome to determine if evolutionary conserved cis-acting packaging signal(s) were preserved after recoding. We showed that conserved synonymous sites and/or local secondary structures that might play a role in determining packaging specificity do not survive codon pair recoding. This makes it unlikely that numerous "cryptic, sequence-degenerate, dispersed RNA packaging signals mapping along the entire viral genome" [Patel N, et al. (2017) Nat Microbiol 2:17098] play the critical role in poliovirus packaging specificity. Considering all available evidence, we propose a two-step assembly strategy for +ssRNA viruses: step I, acquisition of packaging specificity, either (a) by specific recognition between capsid protein(s) and replication proteins (poliovirus), or (b) by the high affinity interaction of a single RNA packaging signal (PS) with capsid protein(s) (most +ssRNA viruses so far studied); step II, cocondensation of genome/capsid precursors in which an array of hairpin structures plays a role in virion formation.


Psychosocial predictors of change in sexual activity and sexual function after hysterectomy in women with pelvic pain.
Potential impact on sexual function is an often-cited concern for many patients considering hysterectomy. The existing literature indicates that sexual function remains stable to slightly improved for most patients who undergo hysterectomy, but most studies demonstrate a small subset of patients in whom sexual function declines after surgery. Unfortunately, there is a lack of clarity as to surgical, clinical, and psychosocial factors that may influence the likelihood of sexual activity after surgery or the magnitude and direction of change in sexual function. Although psychosocial factors are strongly associated with overall female sexual function, there is minimal data exploring the potential impact of these factors on the change in sexual function after hysterectomy. This study aimed to evaluate the relationship between baseline psychosocial factors and both sexual activity and sexual function at 6 months after hysterectomy. Patients undergoing hysterectomy for benign, non-obstetric indications were prospectively recruited as part of an observational cohort study evaluating presurgical predictors of posthysterectomy outcomes on pain, quality of life, and sexual function. The Female Sexual Function Index was administered before hysterectomy and 6 months after surgery. Presurgical psychosocial assessments included validated self-reported measures of depression, resilience, relationship satisfaction, emotional support, and social participation. Complete data was available for 193 patients, of whom 149 (77.2%) reported sexual activity at 6 months after hysterectomy. In the binary logistic regression model examining sexual activity at 6 months, older age was associated with a lower likelihood of sexual activity (odds ratio, 0.91; 95% confidence interval, 0.85-0.96; P=.002). Higher relationship satisfaction before surgery was associated with a greater likelihood of sexual activity at 6 months (odds ratio, 1.09; 95% confidence interval, 1.02-1.16; P=.008). As expected, preoperative sexual activity was associated with a greater likelihood of postoperative sexual activity (odds ratio, 9.78; 95% confidence interval, 3.95-24.19, P<.001). Analyses using Female Sexual Function Index scores were limited to patients who were sexually active at both time points (n=132 [68.4%]). The total Female Sexual Function Index score did not change significantly from baseline to 6 months, but there were statistically significant changes in several individual domains of sexual function. Patients reported significant improvement in desire (P=.012), arousal (P=.023), and pain (P<.001) domains. However, significant decreases were reported in orgasm (P<.001) and satisfaction (P<.001) domains. The proportion of patients who met the criteria for sexual dysfunction was quite high (>60%) at both time points, but there was not a statistically significant change in the proportion from baseline to 6 months. In the multivariate linear regression model, there was no relationship between change in sexual function score and any of the variables examined, including age, endometriosis history, pelvic pain severity, or psychosocial measures. In this cohort of patients with pelvic pain undergoing hysterectomy for benign indications, both sexual activity and sexual function remained fairly stable after hysterectomy. Higher relationship satisfaction, younger age, and preoperative sexual activity were associated with a greater probability of sexual activity at 6 months after surgery. Psychosocial factors, such as depression, relationship satisfaction, and emotional support, and history of endometriosis were not related to change in sexual function among patients who were sexually active both before hysterectomy and at 6 months after surgery.


Measurement of 5-hydroxytryptamine release in the rat medial vestibular nucleus using in vivo microdialysis.
The function of the 5-hydroxytryptamine (5-HT) system in the medial vestibular nucleus (MVN) was evaluated using KCl-evoked 5-HT release assessed by in vivo microdialysis. The release of 5-HT in the MVN was increased by the local application of KCl (100 mM) in a calcium-dependent manner. Likewise, perfusion of a 5-HT reuptake blocking agent, clomipramine (1 microM), increased extracellular levels of 5-HT in the MVN. Finally, pretreatment with the selective 5-HT(1A) agonist, 8-hydroxy-2-(di-N-propylamino) tetralin (1 mg/kg, intraperitoneally), attenuated the KCl-evoked 5-HT release. These findings suggest that 5-HT stored in vesicles is released from nerve terminals in the MVN. In addition, its regulation may be mediated by control of somatodendritic 5-HT(1A) receptor activation.


Eliminating protein oxidation artifacts during High Performance Liquid Chromatography peak fractionation processes.
Therapeutic monoclonal antibodies (mAbs) are inherently heterogeneous and hence generally studied and controlled by an array of orthogonal separation methods. During drug candidate development, fractionation by HPLC is regularly employed to assist peak identification and product understanding. One overlooked challenge is the protein oxidation introduced by the fractionation process. In this study, we report the extent of fractionation-induced protein oxidation, which tends to complicate data interpretation and peak assignments. Higher-energy detectors such as fluorescence detectors and lower fraction concentration were found to exacerbate the oxidation artifacts. Other contributing factors than the detector-induced photostress were also found to contribute significantly to protein oxidation. Furthermore, our study showed that collecting fractions into a solution with oxidation scavengers, such as histidine and methionine, was effective in eliminating the oxidation artifacts introduced by detector exposure and fraction processing steps. Through an example, we demonstrate that the modified fractionation workflow improves the accuracy of peak assignments.


Detection of primary myelofibrosis in blood serum via Raman spectroscopy assisted by machine learning approaches; correlation with clinical diagnosis.
Primary myelofibrosis (PM) is one of the myeloproliferative neoplasm, where stem cell-derived clonal neoplasms was noticed. Diagnosis of this disease is based on: physical examination, peripheral blood findings, bone marrow morphology, cytogenetics, and molecular markers. However, the molecular marker of PM, which is a mutation in the JAK2V617F gene, was observed also in other myeloproliferative neoplasms such as polycythemia vera and essential thrombocythemia. Therefore, there is a need to find methods that provide a marker unique to PM and allow for higher accuracy of PM diagnosis and consequently the treatment of the disease. Continuing, in this study, we used Raman spectroscopy, Principal Components Analysis (PCA), and Partial Least Squares (PLS) analysis as helpful diagnostic tools for PM. Consequently, we used serum collected from PM patients, which were classified using clinical parameters of PM such as the dynamic international prognostic scoring system (DIPSS) for primary myelofibrosis plus score, the JAK2V617F mutation, spleen size, bone marrow reticulin fibrosis degree and use of hydroxyurea drug features. Raman spectra showed higher amounts of C-H, C-C and C-C/C-N and amide II and lower amounts of amide I and vibrations of CH3 groups in PM patients than in healthy ones. Furthermore, shifts of amides II and I vibrations in PM patients were noticed. Machine learning methods were used to analyze Raman regions: (i) 800 cm-1 and 1800 cm-1, (ii) 1600 cm-1-1700 cm-1, and (iii) 2700 cm-1-3000 cm-1 showed 100 % accuracy, sensitivity, and specificity. Differences in the spectral dynamic showed that differences in the amide II and amide I regions were the most significant in distinguishing between PM and healthy subjects. Importantly, until now, the efficacy of Raman spectroscopy has not been established in clinical diagnostics of PM disease using the correlation between Raman spectra and PM clinical prognostic scoring. Continuing, our results showed the correlation between Raman signals and bone marrow fibrosis, as well as JAKV617F. Consequently, the results revealed that Raman spectroscopy has a high potential for use in medical laboratory diagnostics to quantify multiple biomarkers simultaneously, especially in the selected Raman regions.


WITHDRAWN: Can the conventional sextant prostate biopsy reliably diagnose unilateral prostate cancer in low-risk, localized, prostate cancer?
The authors hereby retract the e-publication dated 13 May 2008 and entitled, 'Can the conventional sextant prostate biopsy reliably diagnose unilateral prostate cancer in low-risk, localized, prostate cancer?' The authors are submitting a revised version with the same title. This article's statistics were performed for predicting bilateral prostate cancer outcomes. The article was written to help predict unilateral prostate cancer. Although the statistical numbers are correct, they are backwards. We apologize that the statistics indicate a contrary outcome (eg predicting bilateral cancer instead of unilateral disease).


COVID-19 vaccine behaviors and intentions among a national sample of United States adults ages 18-45.
Vaccination for SARS-CoV-2, the virus that causes COVID-19 illness, is an important public health tool to reduce hospitalizations and deaths. This report focuses on intentions and behaviors related to COVID-19 vaccination among United States (U.S.) adults ages 18-45. From February 25-March 24, 2021, we conducted an online survey assessing COVID-19 vaccine intentions and behaviors, health beliefs, vaccine attitudes, and sociodemographic characteristics. Participants were adults aged 18-45, living throughout the U.S. with oversampling in Florida, panelists of a research panel company directly or via verified partners, and able to read, write, and understand English. Associations between COVID-19 vaccination uptake, intentions, and other study variables were examined through multivariable logistic and proportional odds regression analyses. Among participants in the final analytic sample (n = 2722), 18% reported having received at least one dose of a COVID-19 vaccine. Approximately 31% of unvaccinated participants reported strong intentions to receive a COVID-19 vaccine in the next year, whereas 35% reported strong intentions to receive a COVID-19 vaccine if it were strongly recommended by a healthcare provider. All COVID-19 vaccination outcomes were associated with male gender, sexual minority status, higher levels of education, and previous influenza vaccination. All vaccination intention outcomes were associated with vaccine attitudes and geographic region. Vaccination status and intentions were differentially associated with multiple additional sociodemographic, attitudinal, and/or healthcare experience variables. Several demographic variables, vaccine attitudes, and healthcare experiences were found to contribute to COVID-19 vaccine receipt and intentions. Targeted efforts are necessary to increase uptake of the vaccine in the U.S.


Posttraumatic orbital emphysema: incidence, topographic classification and possible pathophysiologic mechanisms. A retrospective study of 137 patients.
The aim of this study was to retrospectively evaluate the incidence of posttraumatic orbital emphysema (OE) and to propose a radiologic topographic classification as well as a possible pathophysiologic model. Orbital fine-cut (1 mm) computerized tomographic scan slices from 137 patients were used to assess the fracture's type, the presence and position of OE, and periorbital tissue herniation. The OE was categorized into the following 5 compartments: subcutaneous periorbital, peribulbar, retrobulbar extra- and intraconal, and pterygopalatine fossa. The incidence of OE was 61%. OE was more frequently associated with isolated medial wall (78%; P < .001) and combined medial wall/orbital floor (82%; P < .01) fractures. Palpable isolated subcutaneous eyelid emphysema was not related to fracture's type (P = .85). OE suggested medial wall fractures alone or combined with orbital floor fractures. Pathophysiologically, according to the Poiseuille law, the difference in length between the ethmoid and sinusal infundibulii could explain the increased incidence of OE when the medial wall is involved.


Purification of histidine-tagged aequorin with a reactive cysteine residue for chemical conjugations and its application for bioluminescent sandwich immunoassays.
Highly purified histidine-tagged aequorin with a reactive cysteine residue (His-Cys4-aequorin) was obtained from the periplasmic space of Escherichia coli cells by nickel-chelate affinity chromatography and hydrophobic chromatography. The procedure yielded 40.3mg of His-Cys4-aequorin from 2L of cultured cells with over 95% purity. The chemical conjugates of His-Cys4-aequorin with maleimide-activated streptavidin and maleimide-activated biotin were prepared without significant loss of luminescence activity and were applied to the bioluminescent sandwich immunoassay for α-fetoprotein (AFP) as a model analyte. The measurable range of AFP by these conjugates was 0.01-100 ng/ml and the sensitivities were similar to that using aequorin-labeled specific antibody and amino-biotinylated aequorin.


Characterization and physicochemical properties of pectins extracted from agroindustrial by-products.
The composition and fine structure of pectins found in plant cell walls are heterogeneous, with striking differences, depending on their source, and this eventually determines their functional and technological properties. The aim of this study was to extract and determine the chemical composition and physicochemical properties of pectins from different sources: passion fruit peel, orange pomace, and soy hull. Pectin extraction was performed with heated hydrochloric acid solution, followed by precipitation with 96% ethanol. Extraction yield, chemical composition, molar mass, physicochemical properties (fat absorption capacity, cation exchange capacity, water holding capacity, and antioxidant activity) of pectin were measured. Pectin extraction efficiency was higher for passion fruit peel and orange pomace (15.71 and 17.96%, respectively). Soy hull had low pectin extraction (5.66%). Galacturonic acid content was 23.21% for passion fruit peel pectin and 16.01% for orange pomace pectin. Water holding capacity, fat absorption capacity, and cation-binding capacity present in pectin extracted from passion fruit peel were higher, suggesting this poorly investigated product could be used as thickening and emulsifying agents in food preparations. Phenolic compounds with antioxidant capacity provide pectins with additional properties and expand their industrial use.


Transcriptional regulation of the tryptophan oxygenase gene in rat liver by glucocorticoids.
The enzyme tryptophan oxygenase (EC 18.104.22.168), which is synthesized in rat liver, is induced by glucocorticoids. We have used cloned tryptophan oxygenase genomic and cDNA sequences to study the mechanism of induction. Rat liver poly(A+) RNA was separated on a formaldehyde gel, blotted to nitrocellulose, and hybridized to a nick-translated tryptophan oxygenase cDNA clone to analyze the kinetics of tryptophan oxygenase mRNA accumulation. Transcription in isolated rat liver nuclei was investigated to determine the relative rate of transcription of the tryptophan oxygenase gene. Analysis of the accumulation of albumin mRNA, which is unaffected by glucocorticoids, served as an internal control. We show here that the synthetic glucocorticoid dexamethasone causes a 10-fold increase in the concentration of tryptophan oxygenase mRNA sequences in rat liver, and that this is a consequence of transcriptional activation of the tryptophan oxygenase gene.


The case for applying tissue engineering methodologies to instruct human organoid morphogenesis.
Three-dimensional organoids derived from human pluripotent stem cell (hPSC) derivatives have become widely used in vitro models for studying development and disease. Their ability to recapitulate facets of normal human development during in vitro morphogenesis produces tissue structures with unprecedented biomimicry. Current organoid derivation protocols primarily rely on spontaneous morphogenesis processes to occur within 3-D spherical cell aggregates with minimal to no exogenous control. This yields organoids containing microscale regions of biomimetic tissues, but at the macroscale (i.e. 100's of microns to millimeters), the organoids' morphology, cytoarchitecture, and cellular composition are non-biomimetic and variable. The current lack of control over in vitro organoid morphogenesis at the microscale induces aberrations at the macroscale, which impedes realization of the technology's potential to reproducibly form anatomically correct human tissue units that could serve as optimal human in vitro models and even transplants. Here, we review tissue engineering methodologies that could be used to develop powerful approaches for instructing multiscale, 3-D human organoid morphogenesis. Such technological mergers are critically needed to harness organoid morphogenesis as a tool for engineering functional human tissues with biomimetic anatomy and physiology. Human PSC-derived 3-D organoids are revolutionizing the biomedical sciences. They enable the study of development and disease within patient-specific genetic backgrounds and unprecedented biomimetic tissue microenvironments. However, their uncontrolled, spontaneous morphogenesis at the microscale yields inconsistences in macroscale organoid morphology, cytoarchitecture, and cellular composition that limits their standardization and application. Integration of tissue engineering methods with organoid derivation protocols could allow us to harness their potential by instructing standardized in vitro morphogenesis to generate organoids with biomimicry at all scales. Such advancements would enable the use of organoids as a basis for 'next-generation' tissue engineering of functional, anatomically mimetic human tissues and potentially novel organ transplants. Here, we discuss critical aspects of organoid morphogenesis where application of innovative tissue engineering methodologies would yield significant advancement towards this goal.


Modified Candy-plug Device for Dilated False Lumen Occlusion: Is a Candle Better Than a Candy?
The AFX stent graft (Endologix) has an active seal mechanism that can conform to an irregular vascular lumen. We modified the candy-plug technique using an AFX VERA suprarenal extension with restricted purse-string sutures to shape it into a candle-like contraption. We treated 2 patients with patent, dilated false lumens (FLs) after central aortic repair using this technique. This plug was deployed into the FL to block retrograde blood flow proximally. Partial or complete thrombosis of the FL was observed in both patients 6 months after surgery. This technique was feasible for FL occlusion.


Cloning of p97/Gab2, the major SHP2-binding protein in hematopoietic cells, reveals a novel pathway for cytokine-induced gene activation.
Several components in cytokine signaling remain unidentified. We report the cloning and initial characterization of one such component, p97, a widely expressed scaffolding protein distantly related to Drosophila DOS and mammalian Gab1. Upon cytokine, growth factor, or antigen receptor stimulation, p97 becomes tyrosyl phosphorylated and associates with several SH2 domain-containing proteins, including SHP2. Expression of p97 mutants unable to bind SHP2 blocks cytokine-induced c-fos promoter activation, inhibiting Elk1-mediated and STAT5-mediated transactivation. Surprisingly, such mutants do not inhibit MAPK activation. Our results identify p97 as an important regulator of receptor signaling that controls a novel pathway to immediate-early gene activation and suggest multiple functions for SHP2 in cytokine receptor signaling.


Site-directed mutagenesis implicates a threonine residue in TM6 in the subtype selectivities of UH-AH 37 and pirenzepine at muscarinic receptors.
The structural basis for the selectivity of the antagonist UH-AH 37 at human muscarinic acetylcholine receptors was investigated by expressing mutant receptors in COS-7 cells. Previous studies have demonstrated that the interaction between UH-AH 37 and [(3)H]N-methylscopolamine in equilibrium assays is competitive and that the high affinity of UH-AH 37 for the M(5) subtype, compared to M(2), is due to an epitope in the sixth transmembrane domain (TM6) or the third outer loop of the receptor. By mutating each nonconserved residue in this region of M(2) and M(5) to its counterpart in the other receptor, we identified a threonine residue in the middle of TM6 uniquely responsible for the higher affinity of the M(5) receptor (M(1), M(3), and M(4) receptors also carry a threonine at that location and also have high affinity for UH-AH 37). The mutant receptor in which the corresponding alanine of the M(2) receptor was replaced by threonine, M(2)(401)ala --> thr, expressed enhanced affinity for pirenzepine as well as for UH-AH 37. The chick M(2) receptor, which expresses anomalously high affinity for pirenzepine, differs from its mammalian counterparts by the presence of a threonine at this position. Affinities of AF-DX 116 and 4-DAMP, as well as the allosteric potency of UH-AH 37, were not sensitive to the M(2)(401) ala --> thr mutation.


Ischaemia-induced and reperfusion-induced arrhythmias differ in their sensitivity to potassium: implications for mechanisms of initiation and maintenance of ventricular fibrillation.
The effects of four different K+ concentrations (2, 4, 6 or 8 mM) on arrhythmias resulting from ischaemia, and from reperfusion (after 3, 5, 7, 10, 15 or 30 min of ischaemia), in isolated perfused rat hearts were examined. A randomized experimental design with blind analysis was used. Regional myocardial ischaemia was produced by occlusion of the left main coronary artery. The incidence of ischaemia-induced ventricular fibrillation (VF) was inhibited by elevating K+ concentration, as reported previously. Furthermore, this effect was linearly and inversely related to the log of the K+ concentration (r = 0.99, P less than 0.001), a finding which has not been reported previously. This finding implies that the antiarrhythmic effect may result from depolarization in the non-ischaemic tissue, since resting membrane potential is also linearly related to the log of the K+ concentration. The maintenance of ischaemia-induced VF (its tendency to sustain) was also influenced by K+, in that a significantly higher incidence of sustained VF (defined as VF still present at the end of the period of ischaemia) was seen with 2 mM K+ compared with higher K+ concentrations (P less than 0.05). In contrast with its effect on the incidence of ischaemia-induced VF, K+ was without effect on the incidence of reperfusion-induced VF, indicating that reperfusion initiates VF independently of the K+ concentration in the perfusion fluid. However, the maintenance of reperfusion-induced VF was K+-dependent (in a similar manner to the maintenance of ischaemia-induced VF). In summary, the effects of K+ on ischaemia-induced VF were different from its effects on reperfusion-induced VF. We conclude that ischaemia-induced VF and reperfusion-induced VF are unlikely to be initiated by a common electrophysiological mechanism since only the former was influenced by K+. The mechanisms of maintenance of ischaemia-induced VF and reperfusion-induced VF might, however, be common, since the tendency for spontaneous defibrillation to occur (as reflected by the incidence of sustained VF) was equally sensitive to K+ in both settings. Finally, the nature of the time course of susceptibility to ischaemia-induced VF compared with that of reperfusion-induced VF raises the possibility that the reperfusion-induced VF may be clinically relevant as a cause of sudden death only when ischaemia-induced VF is suppressed.


Inadequate social support decreases survival in decompensated liver cirrhosis patients.
Inadequate social support is associated with higher mortality both in the general population and in patients with chronic diseases. There are no studies that have described social support in liver cirrhosis and its impact on prognosis. To analyze the impact social support has in the survival of patients with decompensated cirrhosis. Prospective multicentric cohort study (2016-2019). Patients with decompensated liver cirrhosis were included. Epidemiological, clinical and social variables were collected, using the validated Medical Outcomes Study Social Support Survey, with a 12-month follow-up. A total of 127 patients were included, of which 79.5% were men. The most common etiology of cirrhosis was alcohol (74.8%), mean age was 60 years (SD 10.29), mean MELD was 15.6 (SD 6.3) and most of the patients had a Child-Pugh B (53.5%) or C (35.4%). In the assessment of social support, we observed that most of the patients (92.2%) had adequate global support. At the end of the follow-up (median 314 days), 70.1% of the patients survived. The 1-year survival rate in patients with inadequate global social support was 30%, compared to 73.5% in the presence of social support. In multivariate Cox regression analysis, inadequate social support predicted survival with an adjusted HR of 5.5 (95% CI 2,3-13,4) independently of MELD (HR 1.1, 95% CI 1-1.2), age (HR 1, 95% CI 1-1.1) and hepatocarcinoma (HR 10.6, 95% CI 4.1-27.4). Adequate social support improves survival in liver cirrhosis, independently of clinical variables. Social intervention strategies should be considered for their management.


[Peutz-Jeghers' syndrome with malignant development in a hamartomatous polyp: report of one case and review of the literature].
The malignant potential of hamartomatous polyps in Peutz-Jeghers' (PPJ) syndrome has been debated. Although it is a very rare event, these polyps can become malignant, as demonstrated by this report. One case of colonic adenocarcinoma associated with Peutz-Jeghers' syndrome is described in a 62-year-old woman. The patient had colonic carcinoma which developed in a hamartomatous polyp. The malignant development of this colonic hamartomatous polyp arising in Peutz-Jeghers' syndrome was pathologically confirmed at surgery. This case also shows a sequence of hamartoma-dysplasia-carcinoma in a hamartomatous polyp without adenomatous changes. This suggests that hamartomatous polyps in Peutz-Jeghers' syndrome may develop into adenocarcinoma and may be a precursor of gastrointestinal carcinomas. STK 11 is a tumor suppressor gene regulating the development of hamartomas, and this somatic mutation promotes gastrointestinal cancer at later stages in Peutz-Jeghers' syndrome.