Patients with extreme obesity: change in mental symptoms three years after gastric banding.
Extreme obesity causes grave psychosocial and psychopathological problems in addition to somatic morbidity. One possible treatment is gastric banding, a surgical reduction of stomach volume. The aim of this study was to investigate whether gastric banding leads to lasting change in: 1) the Body Mass Index (BMI); 2) social factors such as work and partnerships, eating behavior, anxiety and depression symptoms; and 3) health related quality of life. We surveyed a sample of 50 adipose women (BMI > 40 kg/m2). Primary outcome measures were self-reported changes on the scales of the Three-Factor Eating Questionnaire (TFEQ), the Hospital Anxiety and Depression Scale (HADS-D), and the Health Survey (SF-36). In comparison with the control group, we observed significant changes in BMI (p < 0.01) and the existence of a partnership (p < 0.01), on all three scales of the TFEQ (p < 0.01), on both scales of the HADS-D (anxiety: p < 0.05; depression:p < 0.01), and on all scales of the SF-36 Health Survey (p between < 0.05 and < 0.01 in every case). The most marked changes in all the qualities investigated occurred within the first 12 months of surgery. Three years after gastric banding, positive changes in BMI reduction, partnership, eating behavior, anxiety, depressive symptomatology, and health related quality of life could be observed. There was also a significant correlation between BMI reduction and reduction firstly on the depression scale (HADS-D) and secondly on the SF-36 scales for physical functioning (PHFU), role physical (ROPH), mental health (PSYC), and vitality (VITA).


Minimally invasive method for murine brain fixation.
Complete brain fixation can be achieved with transthoracic cardiac infusion without thoracotomy. Light and electron microscopy tissue sections reveal preservation of cytoplasmic and nuclear structure at all magnification levels. Punched samples were obtained from the fixed tissue specimens in precisely localized areas for study using electron microscopy. This perfusion fixation technique provides both faster tissue harvesting capability and higher quality tissue preservation, without the artifacts of brain swelling and ventricular dilation observed in direct cardiac perfusion. Acute, discrete change in brain tissue can be studied.


Photoaddition of fluphenazine to nucleophiles in peptides and proteins. Possible cause of immune side effects.
By the action of UVA light, fluphenazine reacted with nucleophiles through a mechanism involving defluorination of its trifluoromethyl group, giving rise to carboxylic acid derivatives that were easily detected by electrospray mass spectrometry. This photoreaction took place with alcohols, sulphydryls, and amines. When irradiation of fluphenazine was carried out in the presence of an amino acid at pH 7.4, the alpha-amino group was covalently bound to the drug. With amino acids possessing a further nucleophilic residue on the side chain, such as lysine, tyrosine, and cysteine--but not serine--both groups reacted, resulting in a fluphenazine-amino acid-fluphenazine diadduct. The same occurred with the physiological peptide glutathione (gamma-glutamylcysteinylglycine). By means of MALDI mass spectrometry, it was shown that fluphenazine also covalently bound to peptides and proteins such as calmodulin. This binding may result in the formation of antibodies, ultimately leading to the destruction of the granulocytes and thus suggesting that photoactivation of this drug may play a role in its clinical side effects, such as agranulocytosis.


[Stress urinary incontinence surgical correction with third generation sub-mid-urethra sling: TVT-secur].
To review our initial experience with the new system TVT-Secur for the stress urinary incontinence. To evaluate the feasibility, efficacy and main advantages of the surgical correction with the new device. Between January and May of 2007, 16 TVT-Secur (Women's Health & Urology, Ethicon, Johnson & Johnson) were made in patients of the Urogynecology Unit, of Clinica Las Condes. Six TVT-Secur were applied in "V" and 10 in "U". The median age was 52 years old, BMI 29, parity of 3. All patients were studied with urodynamic before surgery. All must have the urinary incontinence by more of one year and no must have previous surgical correction. The media surgical time was 10 minutes (8-15 minutes). Complications were not registered during intraoperative and immediate, early or delayed postoperative time. The visual analogue scale of pain was 1 to 2 at 12 hours. All patients were cured. The follow-up was between 1 and 4 months, 2 month media. All patients expressed satisfaction with the surgical results and they would recommend it to other patients in the same conditions. According to our initial experience, the new system TVT-Secur is feasible, safe and effective in the surgical correction of the SUI. However, only the long follow-up and the incorporation of new patients to the study, will allow to determine the permanence of these good results in the time.


Investigation of pyrite oxidation by hexavalent chromium: solution species and surface chemistry.
Pyrite oxidation processes by aqueous Cr(VI) were investigated at 25 degrees C under an argon atmosphere. Synthetic pyrite suspensions (6 g L(-1)) were reacted for 20 h with a range of Cr(VI) solutions from 0 to 700 microM and at pH 2-12. The main objective of this work was to investigate the reaction mechanisms by emphasizing the role of sulfur species. Aqueous chemical processes were well illustrated in acidic media where significant amounts of sulfate and iron species were determined. Sulfate anions are the final stable sulfur species involved in the reaction pathway. Experiments showing complete Cr(VI) removal from solution displayed ratios [S(VI)]/[Fe](tot)<2, probably due to a deficit in aqueous sulfur species. Experiments showing incomplete Cr(VI) removal displayed ratios [Cr(VI)](removed)/[S(VI)] close to 1.5. This ratio was found to be consistent with the formation of thiosulfate (S(2)O(2-)(3)). Thiosulfate ions disproportionated into elemental sulfur S(0) and tetrathionate ions (S(4)O(2-)(6)) that were finally oxidized to sulfate anions under acidic conditions. The distribution of the oxidation state of sulfur atoms at the pyrite surface determined by XPS was additional evidence for the multistep sulfur oxidation process. The presence of elemental sulfur in the S(2p) spectra correlated well with the disproportion of thiosulfate under acidic conditions.


Changes in plasma matrix metalloproteinase-9 levels in patients with acute ischemic stroke.
Matrix metalloproteinases (MMP) have been thought to be involved in stroke pathogenesis. MMP-9 contributes to tissue destruction. Our aim was to analyze the MMP-9 levels in blood within 24 hours of acute ischemic stroke onset to observe the role of MMP-9 in the pathogenesis of atherothrombotic stroke. In this study we investigated prospectively MMP-9 levels in serum from 106 patients (42 men and 64 women, mean age 71.52 +/- 6.32 years) with acute ischemic stroke in the middle cerebral artery area in the first 24 hours from the onset (mean duration 7.8 +/- 4.5 hours) as compared to 112 controls (48 men and 64 women, mean age 70.36 +/- 6.8 years). Serum samples were collected under sterile conditions and stored in aliquots at -70 degrees C until assay. Serum MMP-9 levels were determined by enzyme-linked immunosorbent assay (ELISA) in blood samples obtained on admission. Statistical analysis was performed by Mann-Whitney and Log-Likeliwood Ratio tests. All values reported are expressed as mean (x) +/- SD. Mean serum MMP-9 concentrations were higher in group with ischemic stroke 172 +/- 32.4 ng/mL, range 139.6-204.4 ng/mL vs. controls 57 +/- 9.6 ng/mL, range 47.4-66.6 ng/mL (95% CI, 3.17 to 14.18; p < 0.014). In conclusion, MMP-9 activity is associated with early acute ischemic stroke. The high levels of MMP-9 in acute ischemic stroke document the involvement of this enzyme in the regulation of inflammation in stroke.


Posttranslational modifications in pathogenesis of PCOS.
Polycystic ovary syndrome (PCOS) is a lifelong reproductive, metabolic, and psychiatric disorder that affects 5-18% of women, which is associated with a significantly increased lifetime risk of concomitant diseases, including type 2 diabetes, psychiatric disorders, and gynecological cancers. Posttranslational modifications (PTMs) play an important role in changes in protein function and are necessary to maintain cellular viability and biological processes, thus their maladjustment can lead to disease. Growing evidence suggests the association between PCOS and posttranslational modifications. This article mainly reviews the research status of phosphorylation, methylation, acetylation, and ubiquitination, as well as their roles and molecular mechanisms in the development of PCOS. In addition, we briefly summarize research and clinical trials of PCOS therapy to advance our understanding of agents that can be used to target phosphorylated, methylated, acetylated, and ubiquitinated PTM types. It provides not only ideas for future research on the mechanism of PCOS but also ideas for PCOS treatments with therapeutic potential.


Caudal deficiency sequence in 7q terminal deletion.
We describe a male infant with signs of caudal deficiency sequence. In addition, he showed growth retardation, microcephaly, prominent forehead, bulbous nose tip, large dysplastic ears, hypospadia, partial sacral agenesis, and neurologic bladder dysfunction. Chromosome examination showed a terminal 7q deletion 46,XY,del(7)(pter----q32:). Four previous reported cases of 7q terminal deletion and signs of caudal deficiency are reviewed. Chromosome aberrations may, at least in some cases, be responsible for developmental defects.


When disaster strikes: getting ready for the next big one: part II.
This two-part article deals with the role of the physician when disaster strikes. This second part expands on the role of the physician andpractice in the community and details how the physician and staff can work within the network of the medical societies, hospital systems, government agencies, and others in the event of a far-reaching disaster such as Hurricane Katrina or September 11. It also discusses what the physician can do to educate patients as to how to prepare to meet a disaster.


The Oxford II medial unicompartmental knee arthroplasty: an independent 10-year survival study.
Unicompartmental knee arthroplasty (UKA) is being used increasingly. We report the outcome of a series of 48 Oxford II meniscal bearing UKAs carried out for osteoarthritis of the medial compartment (38 knees in 32 patients) and for spontaneous avascular necrosis of the medial femoral condyle (10 knees in 10 patients). Using the endpoint of revision for any cause, the outcome for every knee was established. Ten knees have been revised (20.8%). At ten years there were 215 knees still at risk and the cumulative survival rate was 77.7% (95% CI: 56.8 to 90.2). Our study reveals a higher failure rate than previous reports on this prosthesis. The technique of implantation of the Oxford knee is demanding; the learning curve is thus long and could explain, at least partially, our moderate results.


Chromosomal localization of the human gene for annexin V (placental anticoagulant protein I) to 4q26----q28.
Annexin V is a member of a new family of calcium-dependent phospholipid-binding proteins. It has been previously isolated as placental anticoagulant protein I, inhibitor of blood coagulation, vascular anticoagulant-alpha, endonexin II, lipocortin V, placental protein 4, and anchorin CII. The human gene encoding annexin V (ANX5) was localized to 4q26----q28 by in situ hybridization with a cDNA probe and polymerase chain-reaction (PCR) analysis of a human x hamster hybrid cell panel. The regional localization to 4q26----q28 was supported by Southern-blot analysis of a human cell line with a deletion in 4q23----q27. This localization overlaps but differs slightly from the previous assignment of ANX5 to 4q28----q32. Digestion with PvuII and TaqI identified polymorphisms at the ANX5 locus; the PvuII polymorphism could also be detected by PCR analysis.


DNA double-strand breaks with 5' adducts are efficiently channeled to the DNA2-mediated resection pathway.
DNA double-strand breaks (DSBs) with 5' adducts are frequently formed from many nucleic acid processing enzymes, in particular DNA topoisomerase 2 (TOP2). The key intermediate of TOP2 catalysis is the covalent complex (TOP2cc), consisting of two TOP2 subunits covalently linked to the 5' ends of the nicked DNA. In cells, TOP2ccs can be trapped by cancer drugs such as etoposide and then converted into DNA double-strand breaks (DSBs) that carry adducts at the 5' end. The repair of such DSBs is critical to the survival of cells, but the underlying mechanism is still not well understood. We found that etoposide-induced DSBs are efficiently resected into 3' single-stranded DNA in cells and the major nuclease for resection is the DNA2 protein. DNA substrates carrying model 5' adducts were efficiently resected in Xenopus egg extracts and immunodepletion of Xenopus DNA2 also strongly inhibited resection. These results suggest that DNA2-mediated resection is a major mechanism for the repair of DSBs with 5' adducts.


Novel Molecular Insights into Classical and Alternative Activation States of Microglia as Revealed by Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC)-based Proteomics.
Microglia, the resident immune cells of the brain, have been shown to display a complex spectrum of roles that span from neurotrophic to neurotoxic depending on their activation status. Microglia can be classified into four stages of activation, M1, which most closely matches the classical (pro-inflammatory) activation stage, and the alternative activation stages M2a, M2b, and M2c. The alternative activation stages have not yet been comprehensively analyzed through unbiased, global-scale protein expression profiling. In this study, BV2 mouse immortalized microglial cells were stimulated with agonists specific for each of the four stages and total protein expression for 4644 protein groups was quantified using SILAC-based proteomic analysis. After validating induction of the various stages through a targeted cytokine assay and Western blotting of activation states, the data revealed novel insights into the similarities and differences between the various states. The data identify several protein groups whose expression in the anti-inflammatory, pro-healing activation states are altered presumably to curtail inflammatory activation through differential protein expression, in the M2a state including CD74, LYN, SQST1, TLR2, and CD14. The differential expression of these proteins promotes healing, limits phagocytosis, and limits activation of reactive nitrogen species through toll-like receptor cascades. The M2c state appears to center around the down-regulation of a key member in the formation of actin-rich phagosomes, SLP-76. In addition, the proteomic data identified a novel activation marker, DAB2, which is involved in clathrin-mediated endocytosis and is significantly different between M2a and either M1 or M2b states. Western blot analysis of mouse primary microglia stimulated with the various agonists of the classical and alternative activation states revealed a similar trend of DAB2 expression compared with BV2 cells.


Storage and secretion of von Willebrand factor by endothelial cells.
Endothelial cells synthesize and store von Willebrand factor. We have studied the storage and secretion of von Willebrand factor in cultured human umbilical vein endothelial cells. In particular, we were interested in the nature of the storage compartment and the effects of perturbation on the storage and secretion processes. The storage compartment for von Willebrand factor was isolated from homogenates of endothelial cells. By an immunostaining technique the isolated vesicles stained for von Willebrand factor. The staining pattern was similar to that of Weibel-Palade bodies in intact endothelial cells. We concluded that the storage compartment containing von Willebrand factor is identical to the Weibel-Palade body. The von Willebrand factor of the isolated storage vesicles is predominantly constructed of polypeptide chains with a Mr of 220 kD. On the other hand, von Willebrand factor continuously secreted by endothelial cells is constructed of both a 220 kD and a larger precursor (apparent Mr of 275 kD) subunit. The storage vesicles contain von Willebrand factor that supports ristocetin-induced platelet aggregation. Thus, endothelial cells store fully processed, biologically active von Willebrand factor within Weibel-Palade bodies. Short-term (less than 1 h) treatment of endothelial cells with the perturbing phorbol ester 4 beta-phorbol-12-myristate-13-acetate (PMA) results in release of cellular stored von Willebrand factor. 24-48 h after exposure to PMA the endothelial cell distribution of von Willebrand factor is changed distinctly. While the contents of the von Willebrand factor storage sites in the cells are gradually restored within 48 h, enhanced amounts of von Willebrand factor are secreted into the medium. The number as well as the size of von Willebrand factor storage granules in the endothelial cells increase after exposure to phorbol ester, as determined by immunofluorescence microscopy. Phorbol ester treated cells release stored von Willebrand factor 48 h after they have been stimulated. PMA decreases the von Willebrand factor contents of the extracellular matrix; the deposition of von Willebrand factor in the subendothelium is blocked by PMA, whereas the degradation of matrix von Willebrand factor is not affected. Thus, perturbation of endothelial cells changes the cellular distribution of von Willebrand factor.


The Spine painDETECT questionnaire: Development and validation of a screening tool for neuropathic pain caused by spinal disorders.
To develop screening tools for neuropathic pain caused by spinal disorders, the Spine painDETECT questionnaire (SPDQ) and its short-form version (SF-SPDQ), by modifying the Japanese version of the painDETECT questionnaire (PDQ-J), and to validate these tools. Using data from patients with neuropathic pain caused by spinal disorders (NeP-SD) and patients with nociceptive pain caused by joint disorders (NocP) as controls, we devised a scoring system for the SPDQ by calculating weighting coefficients for nine PDQ-J items. Simultaneously, we selected some items for the SF-SPDQ. Next, we conducted the validation study primarily using patients with a confirmed diagnosis (a multicenter study) and general patients (a web-based survey). Sensitivity, specificity, and the area under the receiver-operating characteristic curve (AUC), along with additional positive/negative predictive values and positive/negative likelihood ratios, were calculated to assess the diagnostic utility of these tools in each population. Data for 85 patients with NeP-SD and 45 patients with NocP were analyzed to develop the SPDQ/SF-SPDQ. The SPDQ had sensitivity of 78.8% and specificity of 75.6% (AUC = 0.77). The SF-SPDQ had 82.4% sensitivity and 66.7% specificity (AUC = 0.75). In the multicenter study (n = 45), both tools had diagnostic utility almost comparable with that demonstrated at development: the SPDQ had sensitivity of 83.3% and specificity of 69.2%, with the SF-SPDQ having 86.2% sensitivity and 68.8% specificity. In the web-based survey (n = 500), while the SPDQ had slightly low sensitivity (74.0%), the SF-SPDQ maintained high sensitivity (84.4%), although specificity was relatively low (61.2%). We developed the SPDQ and SF-SPDQ as valid screening tools for neuropathic pain caused by spinal disorders. Both have moderate utility as screening tools, with the SF-SPDQ perhaps being preferable for clinical use. However, physicians should be vigilant about possible false-positive diagnoses.


The prostate-specific antigen test.
Before the 1980s, prostate cancer (PC) was considered a deadly disease. The mortality-incidence ratio showed that 1 out of each 2 - 3 PC patients died of this disease. On the other hand, autopsy studies have shown that latent PC is common in middle-aged men. The prostate-specific antigen (PSA), a glycoprotein produced by the epithelial cells of the prostate gland, received FDA's approval in 1986 for monitoring treatment response, and in 1994 as a screening aid for the diagnosis of PC. After the publication of two randomized trials on PC screening using the PSA test, it is generally accepted that systematic PSA-based screening, as compared to a clinical situation with virtually no screening, can reduce suffering from metastatic disease and PC mortality. However, what is also shown is that PSA-based screening coincides with a considerable amount of unnecessary testing and overdiagnosis. Should we abandon the use of the PSA test for the diagnosis of PC, or should we encourage PSA testing and make it freely available for all men at any time? Both the answers should be "No." What we must do is use the test as wisely as is currently possible and inform men, who want to be tested, in a balanced way about harms and potential benefits.


Establishment and verification of a digital PCR assay for the detection of CK19 expression in quantitative analysis of circulating tumor cell.
The objective of this study was to establish and verify a digital PCR assay for the detection of CK19 gene expression, and to use it to detect circulating tumor cells (CTC) by taking advantages of its ultra-high sensitivity and absolute quantitation. Firstly, the primers and probes were designed according to the mRNA sequence of CK19 gene, and housekeeping gene ABL1 was used as the internal control. The best candidate was screened by human breast cancer MCF7 cells and healthy human leukocytes from 13 sets of primer and probes and verified by direct sequencing. Secondly, after the reaction conditions of the selected primers and probes were optimized, limit of blank (LOB) analysis were performed with different concentrations of cDNAs as templates from healthy human leukocytes. The results revealed the LOB of CK19 with ABL1 copy numbers of 20,000, 15,000, 10,000, 5000 and 2500 were 9.24, 8.93, 3.12, 3.17 and 2.53 copies, respectively. Thirdly, the different concentrations of cDNAs from MCF7 cells and healthy human leukocytes were premixed and used in the limit of detection (LOD) analysis, which showed that the CK19 gene could be effectively detected at the concentration ratio of 50%, 10%, 5%, 1%, 0.5% and 0.1%, and the linear R2 value was 0.9998. Finally, the preliminary results of digital PCR in clinical samples indicated that CK19 copy numbers were higher in advanced breast cancer patients than healthy controls. The above results demonstrated the advantages of our CK19 digital PCR assay in sensitivity, specificity, and accurate quantification. If verified further, the assay is expected to play significant roles in the quantitative analysis of CTC in breast cancer with a good application prospect.


MicroRNA-mediated posttranscriptional mechanisms of gene expression in proliferating and quiescent cancer cells.
MicroRNAs are small non-coding RNA regulators of gene expression that play important roles in critical biological processes, including cell division, self-renewal and cell state maintenance. Their deregulation leads to extensive clinical consequences in tumorigenesis. Cancers demonstrate heterogeneity in their cell states implicated in their resistance and resurgence. Apart from proliferating cells, cancers harbor a small proportion of assorted quiescent cells that resist conventional therapeutics and contribute to cancer recurrence. MicroRNA expression, targets, microRNPs (microRNA-protein complexes) and their functions have been demonstrated to be regulated in distinct tumor cell states and as an adaptive response to stress signals in tumor-unfavorable environments. In turn, altered microRNPs and their modified post-transcriptional mechanisms of gene expression may contribute to tumor resistance and influence tumor progression. An understanding of distinct microRNA mechanisms in cancer cells would provide extensive insights into the versatile roles of microRNAs in the perpetuation of tumors and indicate potential therapeutic avenues.


Sensory deprivation changes the pattern of synaptic connectivity in rat barrel cortex.
We examined whether sensory deprivation during formation of the cortical circuitry influences the pattern of intracortical single-cell connections in rat barrel cortex. Excitatory postsynaptic potentials from layer 5 pyramidal neurons were recorded in vitro using patch-clamp techniques. In order to evoke such postsynaptic potentials presumptive presynaptic neurons were stimulated by photolytically applied glutamate thus generating action potentials. Synaptic connections between the stimulated and the recorded neuron were identified by the occurrence of postsynaptic potentials following photostimulation. Sensory deprivation altered the projections from layer 2/3 neurons to layer 5 pyramidal cells (L2/3-->L5 projections). In slices of non-deprived rats the input probability of L2/3-->L5 projections showed a periodic pattern with more synaptic connections originating from the borders of the barrel columns, and less synaptic connections originating from the centres. After whisker clipping this periodic pattern disappeared completely and the input probability declined monotonically with increasing distance between stimulated and recorded neuron. These results indicate that sensory input is a prerequisite to establish a synaptic projection pattern which is correlated to the columnar organisation of the anatomical barrel structure.


Sharpness-enhanced ultrafast imaging by using a biased optical Kerr gate.
We proposed a novel biased optical Kerr gated imaging (BOKGI) method for ultrafast imaging. The imaging performance of the BOKGI system has been investigated. Experimental results showed that by using the BOKGI, the high spatial frequency components of the detected object could be effectively retrieved, which are often filtered by the photo-induced soft aperture in a conventional OKGI system. Comparing with the conventional OKGI method, the BOKGI method could enhance the sharpness of images and provide a higher spatial resolution of the imaging system. In addition, the influence of the biased angle on the BOKGI performance has been also investigated.


Hemiface Differences in Visual Exploration Patterns When Judging the Authenticity of Facial Expressions.
Past studies have found asymmetry biases in human emotion recognition. The left side bias refers to preferential looking at the left-hemiface when actively exploring face images. However, these studies have been mainly conducted with static and frontally oriented stimuli, whereas real-life emotion recognition takes place on dynamic faces viewed from different angles. The aim of this study was to assess the judgment of genuine vs. masked expressions in dynamic movie clips of faces rotated to the right or left side. Forty-eight participants judged the expressions on faces displaying genuine or masked happy, sad, and fearful emotions. The head of the actor was either rotated to the left by a 45° angle, thus showing the left side of the face (standard orientation), or inverted, with the same face shown from the right side perspective. The eye movements were registered by the eye tracker and the data were analyzed for the inverse efficiency score (IES), the number of fixations, gaze time on the whole face and in the regions of interest. Results showed shorter IESs and gaze times for happy compared to sad and fearful emotions, but no difference was found for these variables between sad and fearful emotions. The left side preference was evident from comparisons of the number of fixations. Standard stimuli received a higher number of fixations than inverted ones. However, gaze time was long on inverted compared to standard faces. Number of fixations on exposed hemiface interacted with the emotions decreasing from happy to sad and fearful. An opposite pattern was found for the occluded hemiface. These results suggest a change in fixation patterns in the rotated faces that may be beneficial for the judgments of expressions. Furthermore, this study replicated the effects of the judgment of genuine and masked emotions using dynamic faces.


Color stability of 3D-printed denture resins: effect of aging, mechanical brushing and immersion in staining medium.
This in-vitro study evaluated and compared the color stability of 3D-printed and conventional heat-polymerized acrylic resins following aging, mechanical brushing, and immersion in staining medium. Forty disc-shaped specimens (10 mm in diameter and 3 mm thick) were prepared from two 3D-printed [DentaBASE (DB) and Denture 3D+ (D3D)] and one conventional polymethylmethacrylate (PMMA) denture materials. The specimens were thermo-cycled, subjected to mechanical brushing, and were immersed in either coffee, lemon juice, coke, or artificial saliva (AS) to simulate one and two years of oral use. Color measurements of the specimens were recorded by a spectrophotometer at baseline (T0), and after one (T1) and two years (T2) of simulation. The color changes (∆E) were determined and also quantified according to the National Bureau of Standards (NBS) units. Descriptive statistics, followed by factorial ANOVA and Bonferroni post-hoc test (α=.05), were applied for data analysis. The independent factors, namely material, staining medium, and immersion time, and interaction among these factors significantly influenced ∆E (P<.009). Irrespective of the materials, treatments, and time, the highest and the lowest mean ∆Es were observed for PMMA in lemon juice (4.58 ± 1.30) and DB in AS (0.41 ± 0.18), respectively. Regarding the material type, PMMA demonstrated the highest mean ∆E (2.31 ± 1.37), followed by D3D (1.67 ± 0.66), and DB (0.85 ± 0.52), and the difference in ∆E between the materials were statistically significant (P<.001). All the specimens demonstrated a decreased color changes at T2 compared to T1, and this difference in mean ∆E was statistically significant (P<.001). The color changes of 3D-printed denture resins were low compared to conventional heat polymerized PMMA. All the tested materials, irrespective of the staining medium used, demonstrated a significant decrease in ∆E values over time.


Biochemical characterization of angiotensin-converting enzyme in human neuroblastoma tissue.
High activity of angiotensin-converting enzyme was demonstrated in human neuroblastoma tissue. This activity required the presence of chloride ion and was almost completely inhibited by a specific converting enzyme inhibitor captopril (10 nM), indicating that the activity measured is indeed angiotensin-converting enzyme. Furthermore, the biochemical features of the enzyme were closely similar to the well-known properties of human lung converting enzyme, such as molecular weight (290,000), optimum pH (8.0-8.5), the presence of glycoprotein residues, and dependence on chloride ion concentration. These results provide definitive evidence for the presence of true angiotensin-converting enzyme in human neuroblastoma tissue.


Prognosis in schizophrenia. A ten-year follow-up of first admissions.
Eighty-eight of 92 first admission schizophrenics from 1963, being an incidence by first admission cohort, were followed up between 1974 and 1975. Fifty-eight percent were reported as showing no social or intellectural deficit, and only 8% were unremitting institutionalized, with 51% being considered as having normal economic productivity, and 69% with a good or fair social adjustment. Despite difficulties in comparison, birth, marriage, and divorce rates appear to be not very different from those for the general population, although the percentage of married patients is lower. Deaths would seem greater than expected. Possible reasons for the improved prognosis in schizophrenia are considered to be short initial hospitalization, almost universal use of phenothiazines, use of developing community services (social and psychiatric), and generally good econimic conditions.


Request for euthanasia by a psychiatric patient with undetected intellectual disability.
In the Netherlands, euthanasia or assisted suicide (EAS) in psychiatric disorders is legal in certain circumstances. Guidelines recommend a second opinion to independently check diagnosis and treatment resistance. A 68-year-old patient, diagnosed with bipolar I disorder, with a request for euthanasia because of tiredness, repeated falls and racing thoughts was seen for such a second opinion. Persisting in her wish, her reluctant family and psychiatrist became convinced of euthanasia. Our disagreement with the diagnosis of bipolar I disorder upset her, but she agreed with discontinuation of psychotropic drugs. Her mobility and tiredness improved, whereafter her request for euthanasia evolved into a death wish due to completed life. Intellectual disability and an attention deficit hyperactivity disorder could explain her struggle in life. This case report shows that extending the procedure regarding EAS with an independent psychiatric evaluation is important. For our patient, this second opinion supported her to find meaning in life.


Glycomic and glycoproteomic analysis of serum from patients with stomach cancer reveals potential markers arising from host defense response mechanisms.
Despite the reduced incidence of gastric cancer in the developed world, a diagnosis of stomach carcinoma still carries a poor prognosis due to the asymptomatic nature of the disease in the early stages, subsequent advanced stage diagnosis, and a low 5 year survival rate. Endoscopy remains the primary standard for diagnosis of stomach carcinoma and the current marker, carbohydrate antigen 19-9 (CA19-9) lacks the levels of sensitivity and specificity required in order to make it clinically useful for diagnostic monitoring. Therefore, there is a current need for additional markers to improve the diagnostic accuracy for the early stages of stomach cancer. Together, glycomic, proteomic, and glycoproteomic analyses of serum have the potential to identify such probable markers. A discovery study is reported here using preoperative serum from 80 stomach cancer patients, 10 patients bearing benign stomach disease, and 20 matched controls. Glycomic analysis of the total and immunoaffinity depleted serum revealed statistically significant increases in the levels of sialyl Lewis X epitopes (SLe(X)) present on triantennary glycans accompanied by increased levels of core fucosylated agalactosyl biantennary glycans present on IgG (referred to as the IgG G0 glycoform) which are associated with increasing disease pathogenesis. Protein expression analysis using 2D-DiGE returned a number of differentially expressed protein candidates in the depleted serum, many of which were shown to carry triantennary SLe(X) during subsequent glycomic investigations. Biological pathway analysis of the experimental data returned complement activation and acute phase response signaling as the most significantly altered pathways in the stomach cancer patient serum. Upon the basis of these findings, it is suggested that increased expression of IgG G0 and complement activation are a host response to the presence of the stomach tumor while the increased expression of SLe(X) and acute phase response proteins is a result of pro-inflammatory cytokine signaling, including IL-6, during carcinogenesis. The approach presented herein provides an insight into the underlying mechanisms of disease and the resulting changes in the glycome and glycoproteome offer promise as potential markers for diagnosis and prognostic monitoring in stomach cancer.


Erythroplasia of Queyrat with Zoon's balanitis: a diagnostic dilemma.
Chronic erythematous lesions of the penis may result from a variety of underlying causes that form a part of differential diagnosis. They are difficult to diagnose only upon clinical examination and may necessitate performing a biopsy to confirm the diagnosis and also identify the coexisting disease. We report a case of erythroplasia of Queyrat (EQ) with Zoon's balanitis and discuss the classification of the lesion through the analysis of overlapping histological features. A middle-aged uncircumcised man presented with two long-standing erythematous prepucial penile lesions unresponsive to antibiotics. Biopsy performed to establish the diagnosis revealed non-invasive severe dysplastic changes in the epithelium that is typical of EQ. Subepithelial histological features were characteristic of Zoon's balanitis. The extent and nature of inflammatory infiltrate in the dermis described in the literature is quite varied in EQ and in the spectrum of non-cicatricial balanoposthitis. The overlap of histological findings could result in the diagnostic dilemma of a coexistent lesion, as described in this case report.


Spontaneously amplified homochiral organic-inorganic nano-helix complexes via self-proliferation.
Most spiral coiled biomaterials in nature, such as gastropod shells, are homochiral, and the favoured chiral feature can be precisely inherited. This inspired us that selected material structures, including chirality, could be specifically replicated into the self-similar populations; however, a physicochemical understanding of the material-based heritage is unknown. We study the homochirality by using calcium phosphate mineralization in the presence of racemic amphiphilic molecules and biological protein. The organic-inorganic hybrid materials with spiral coiling characteristics are produced at the nanoscale. The resulted helixes are chiral with the left- and right-handed characteristics, which are agglomerated hierarchically to from clusters and networks. It is interesting that each cluster or network is homochiral so that the enantiomorphs can be separated readily. Actually, each homochiral architecture is evolved from an original chiral helix, demonstrating the heritage of the matrix chirality during the material proliferation under a racemic condition. By using the Ginzburg-Landaue expression we find that the chiral recognition in the organic-inorganic hybrid formation may be determined by a spontaneous chiral separation and immobilization of asymmetric amphiphilic molecules on the mineral surface, which transferred the structural information from the mother matrix to the descendants by an energetic control. This study shows how biomolecules guide the selective amplification of chiral materials via spontaneous self-replication. Such a strategy can be applied generally in the design and production of artificial materials with self-similar structure characteristics.


Regional generalisations about the relationships between the environment and foraminifera along the SW Cape coast, South Africa.
Factors influencing the composition of shallow water benthic foraminifera along the SW coast of South Africa at two locations (Table Bay, TB; St Helena Bay, SHB) ∼200km from each other were examined. Small taxa dominated in both locations; living assemblages from SHB (28 species, Ammonia parkinsoniana dominant) differed from TB (34 species, Elphidium articulatum dominant). Environmental parameters were similar in both areas. Patchiness in assemblage structure was pronounced, differences between pipeline and non-pipeline sites within locations were evident in environmental parameters. Diversity was significantly correlated with grain size, the nitrogen and heavy metal content (especially Cd). These data represent the first for extant benthic Foraminifera from the southern Benguela upwelling area and is the first attempt at using foraminifera as indicators of pollution in this region. The data stressed the need for more than one sampling location to better understand the factors influencing foraminiferal assemblages in any regional context.


Protection against sucrose-induced retinal capillary damage in the Wistar rat.
The procedure of rearing rats on high sucrose diets has been used in some laboratories as a drug-free model to induce retinal vascular-bed damage resembling diabetic retinopathy. Examination of dietary salt mixes used in laboratories reporting success with this model revealed that two essential elements, chromium and selenium, were not included. The present study evaluated the effect of supplementing a high sucrose diet, reported to induce retinopathy, with chromium and Selenium supplementation significantly reduced the number of acellular collapsed capillary strands within arterial-venous capillary networks. The addition of chromium to the high sucrose diet resulted in lower fasting blood glucose and a smaller rise in its level following an oral dose. Little or no retinal vascular damage was apparent in rats fed the high sucrose diet supplemented with chromium, selenium and additional corn oil. Of the two elements, only supplementation with Se protected the retinal capillaries from damage.


Imaging tumor angiogenesis: the road to clinical utility.
Tumor growth and progression require the formation of new blood vessels from preexisting vasculature, a process called angiogenesis. The ability to noninvasively visualize angiogenesis may provide new opportunities to more appropriately select patients for antiangiogenesis treatment and to monitor treatment efficacy. The superior molecular sensitivity of PET and the lack of radiation from MRI and contrast-enhanced ultrasound put these techniques at the forefront of clinical translation.


Neonatal Vocal Fold Paralysis.
Vocal fold paralysis (VFP) is an important cause of respiratory and feeding compromise in infants. The causes of neonatal VFP are varied and include central nervous system disorders, birth-related trauma, mediastinal masses, iatrogenic injuries, and idiopathic cases. Bilateral VFP often presents with stridor or respiratory distress and can require rapid intervention to stabilize an adequate airway. Unilateral VFP presents more subtly with a weak cry, swallowing dysfunction, and less frequently respiratory distress. The etiology and type of VFP is important for management. Evaluation involves direct visualization of the vocal folds, with additional imaging and testing in select cases. Swallowing dysfunction, also known as dysphagia, is very common in infants with VFP. A clinical assessment of swallowing function is necessary in all cases of VFP, with some patients also requiring an instrumental swallow assessment. Modification of feeding techniques and enteral access for feedings may be necessary. Airway management can vary from close monitoring to noninvasive ventilation, tracheostomy, and laryngeal surgery. Long-term follow-up with otolaryngology and speech-language pathology service is necessary for all children with VFP to ensure adequate breathing, swallowing, and phonation. The short- and long-term health and quality-of-life consequences of VFP can be substantial, especially if not managed early.


Realizing Efficient Emission in Three-Dimensional CsCdCl3 Single Crystals by Introducing Separated Emitting Centers.
Hitherto, three-dimensional (3D) perovskite single crystals with a low exciton binding energy generally possess inferior photoluminescence (PL) performance due to the spatially unconfined nature of excitons. In this work, 3D CsCdCl3 single crystals with multiple emissions from self-trapped excitons (STEs) have been developed, which unsurprisingly exhibit a discouraging PL quantum yield (PLQY) of ∼4.8%. To improve the luminescence efficiency, Mn2+ and Sn2+ are introduced into the lattice as dopants, respectively. By embedding Mn2+ ion into CsCdCl3, the long Mn-Mn distance enables the resultant material to produce an intense orange emission (∼100% PLQY) from the d-d orbital transition (4T1-6A1) of Mn2+. Intriguingly, the embedded Sn2+ triggers the formation of Jahn-Teller-like STEs that induces a subsequent deep red emission with a PLQY of ∼28.22%, which is quite high for 3D bulk perovskites. Such a remarkable PL efficiency is attributed to the distinctive bonding mode of CsCdCl3 that encourages the expression of the Sn 5s2 lone pair. Furthermore, a white-light-emitting diode (WLED) is also fabricated with Mn2+-doped CsCdCl3 to show its potential in lighting application. This work paves a new avenue to improve the luminescence performance of bulk 3D perovskite materials.


Breast cancer.
This synthesis of the literature on radiotherapy for breast cancer is based on 97 scientific articles, including 5 meta-analyses, 38 randomized studies, and 27 retrospective studies. These studies involve 387634 patients. Radiotherapy is the most effective method for preventing locoregional recurrence following primary surgery for invasive breast cancer, and radiotherapy is currently more effective than adjuvant chemotherapy after either mastectomy or breast-conserving surgery. Radiotherapy in patients at high risk for locoregional recurrence, eg, patients with spread to the axillary lymph nodes, leads to a significant increase in relapse free survival (eg, 1, 3). Meta-analyses have shown that radiotherapy in these subgroups of patients can reduce the risk for distant metastasis and reduce the risk for cancer death. These analyses have not statistically confirmed an improvement in total survival, probably because reduced mortality from breast cancer has been offset by increased mortality from cardiovascular disease. However, the results have successively improved, and survival gains are significantly greater in recent studies using modern treatment methods. It is probable that survival gains from radiotherapy do not exceed those that can be achieved by other adjuvant treatment of breast cancer such as chemotherapy or hormones, ie, a reduction in mortality by 20% to 30%, leading to an increased total survival after, eg, 10 years of 5% to 10%. The heart is the most important organ at risk during radiotherapy for breast cancer. Minimizing radiation doses to the heart muscle and the coronary arteries is necessary for avoiding later effects of ischemic cardiovascular disease. These side effects were particularly prominent in early treatment studies that used older radiotherapy methods. Radiotherapy in conjunction with breast-conserving surgery for invasive breast cancer significantly reduces the recurrence frequency in the breast. Clinical studies are under way that aim at further defining the role of radiotherapy as an element in a breast-conserving treatment strategy, e.g., determining the value of boost, and identifying prognostic/predictive factors for breast recurrence. Improved knowledge about such factors should eventually permit identification of patient groups at such low risk for breast recurrence that routine radiotherapy is unnecessary, or at such high risk-even with radiotherapy-that alternatives to breast conserving surgery should be considered. Radiotherapy also reduces the risk for recurrence in the breast following breast-conserving surgery of DCIS. Controlled trials are under way that aim at more closely defining the roles of surgical methods and radiotherapy for various subgroups of patients, eg, regarding different histopathologic types of DCIS. Radiotherapy has a substantial palliative value to patients who cannot be cured. It can reduce, prevent, or delay unpleasant symptoms from advanced disease, eg, pain, cancer lesions, fractures, neurologic symptoms. etc.


'Craziness' and creativity: Psychopathology and Poetry.
Not all poets have experienced psychopathology. Conversely, not all those who have experienced psychopathology become poets. The notion, nonetheless, of there being an association between 'craziness' and creativity, contentious though it may be, remains a seductive one. Poetry is both beneficial for the person who is composing or reciting it as well as the person who may be reading or listening to it. Poetry Therapy, which falls under the remit of Art Therapy, is increasingly being recognised as an effective form of adjunctive therapy for the treatment of mental health problems. The main aims of this paper are to explore (and to attempt to elucidate) if there is indeed a relationship between the artistic temperament and mental illness and to comment on the rise and recognition of Art Therapy.


Identification of cell senescence molecular subtypes in prediction of the prognosis and immunotherapy of hepatitis B virus-related hepatocellular carcinoma.
Hepatitis B virus (HBV)-infected hepatocellular carcinoma (HCC) has a high incidence and fatality rate worldwide, being among the most prevalent cancers. The growing body of data indicating cellular senescence (CS) to be a critical factor in hepatocarcinogenesis. The predictive value of CS in HBV-related HCC and its role in the immune microenvironment are unknown. To determine the cellular senescence profile of HBV-related HCC and its role in shaping the immune microenvironment, this study employed a rigorous evaluation of multiple datasets encompassing 793 HBV-related HCC samples. Two novel distinct CS subtypes were first identified by nonnegative matrix factorization, and we found that the senescence-activated subgroup had the worst prognosis and correlated with cancer progression. C1 and C2 were identified as the senescence-suppressed and senescence-activated subgroups. The immune microenvironment indicated that C2 exhibited a relatively low immune status, higher tumor purity, and lower immune scores and estimated scores, while the C1 subgroup possessed a better prognosis. The CS score signature based on five genes (CENPA, EZH2, G6PD, HDAC1, and PRPF19) was established using univariate Cox regression and the lasso method. ICGC-LIRI and GSE14520 cohorts were used to validate the reliability of the CS scoring system. In addition, we examined the association between the risk score and hallmark pathways through gene set variation analysis and gene set enrichment analysis. The results revealed a high CS score to be associated with the activation of cell senescence-related pathways. The CS score and other clinical features were combined to generate a CS dynamic nomogram with a better predictive capacity for OS at 1, 2, and 3 years than other clinical parameters. Our study demonstrated that cellular senescence patterns play a non-negligible role in shaping the characteristics of the immune microenvironment and profoundly affecting tumor prognosis. The results of this study will help predict patient prognosis more accurately and may assist in development of personalized immunotherapy for HBV-related HCC patients.


Design and operation of a portable micro-photoluminescence spectrometer for education on semiconductor quantum structures and graphene sheets.
The design and operation of a portable micro-photoluminescence spectrometer for applications in education is described. Guidelines are a compact, robust, portable, and flexible design; operation without cryogenic media for sample cooling; and a limited budget. Targeted samples are semiconductor quantum structures emitting in a wavelength range of 600-1000 nm and graphene sheets. The portable spectrometer includes a reflected-light microscope with a motorized sample stage of 156 nm step size, a thermoelectric sample cooler allowing temperatures down to 196 K, a green and a blue laser for focused excitation, a monochromator with 0.18 nm spectral resolution, and a cooled camera as the image sensor. For demonstration of the capabilities of the spectrometer, measurements of the quantized energy levels of molecular beam epitaxy grown GaAs quantum dots (QDs) are shown. Here, different sample designs are used, the sample temperature as well as the laser excitation power and energy is varied, and the respective influence on the measurements is discussed. A clear QD shell structure with four states is shown for a sample, where approximately four QDs are directly excited by a focused laser. Limitations of the spectrometer for QD characterization mainly due to the waiver of cryogenic media for sample cooling are discussed. As a further example, which does not require sample cooling, local Raman spectroscopy of a graphene sheet is demonstrated where clear Raman signatures allow the identification of a single-layer thickness.


Sibling Relations in Patchwork Families: Co-residence Is More Influential Than Genetic Relatedness.
In "patchwork" families, full siblings, maternal and paternal half-siblings, and non-related children are raised together, and sometimes, genetically related children are separated. As their number is steadily growing, the investigation of the factors that influence within-family relations is becoming more important. Our aim was to explore whether people differentiate between half- and full-siblings in their social relations as implied by the theory of inclusive fitness, and to test whether co-residence or genetic relatedness improves sibling relations to a larger extent. We administered the Sibling Relationship Questionnaire to 196 individuals who were in contact with full-, half-, or step-siblings in their childhood. We built Generalized Linear Mixed Models models to test for the effects of relatedness and co-residence on sibling relations. In general, a higher degree of relatedness was associated with better sibling relations, but only among those who did not live together during childhood. Co-resident siblings' overall pattern of relation quality was not influenced by the actual level of genetic relatedness. In contrast to this, full siblings reported having experienced more conflicts during childhood than half-siblings, possibly resulting from enhanced competition for the same parental resources. The results suggest that inclusive fitness drives siblings' relations even in recent industrial societies. However, among individuals who live together, the effect of relatedness might be obscured by fitness interdependence and the subjective feeling of kinship.


Effect of Endosymbiotic Bacteria on Fungal Resistance Toward Heavy Metals.
Most studies on metal removal or tolerance by fungi or bacteria focus on single isolates, without taking into consideration that some fungi in nature may be colonized by endobacteria. To address this knowledge gap, we investigated the tolerance and removal of diverse metals with two fungal species: Linnemannia elongata containing Burkholderia-related endobacteria and Benniella erionia containing Mollicute-related endobacteria. Isogenic lines of both species were generated with antibiotic treatments to remove their respective endobacteria. Experiments involved comparing the isogenic lines and wild type fungi in relation to the minimum inhibitory concentration for the metals, the fungal ability to remove these different metals via atomic adsorption spectroscopy, and the interaction of the metals with specific functional groups of the fungi and fungi-bacteria to determine the role of the bacteria via attenuated total reflection fourier transformed infrared (ATR-FTIR). Finally, we determined the influence of different metal concentrations, associated with moderate and high fungal growth inhibition, on the presence of the endobacteria inside the fungal mycelium via quantitative real-time PCR. Results showed that the presence of the endosymbiont increased B. erionia resistance to Mn2+ and increased the removal of Fe2+ compared to isogenic lines. The absence of the endosymbiont in L. elongata increased the fungal resistance toward Fe2+ and improved the removal of Fe2+. Furthermore, when the bacterial endosymbiont was present in L. elongata, a decrease in the fungal resistance to Ca2+, Fe2+, and Cr6+was noticeable. In the ATR-FTIR analysis, we determined that C-H and C = O were the major functional groups affected by the presence of Cu2+, Mn2+, and Fe2+ for L. elongata and in the presence of Cu2+ and Ca2+ for B. eronia. It is noteworthy that the highest concentration of Pb2+ led to the loss of endobacteria in both L. elongata and B. eronia, while the other metals generally increased the concentration of endosymbionts inside the fungal mycelium. From these results, we concluded that bacterial endosymbionts of fungi can play a fundamental role in fungal resistance to metals. This study provides the first step toward a greater understanding of symbiotic interactions between bacteria and fungi in relation to metal tolerance and remediation.


Real-time Precise Point Positioning with a Xiaomi MI 8 Android Smartphone.
The Global Navigation Satellite System (GNSS) positioning technology using smartphones can be applied to many aspects of mass life, and the world's first dual-frequency GNSS smartphone Xiaomi MI 8 represents a new trend in the development of GNSS positioning technology with mobile phones. The main purpose of this work is to explore the best real-time positioning performance that can be achieved on a smartphone without reference stations. By analyzing the GNSS raw measurements, it is found that all the three mobile phones tested have the phenomenon that the differences between pseudorange observations and carrier phase observations are not fixed, thus a PPP (precise point positioning) method is modified accordingly. Using a Xiaomi MI 8 smartphone, the modified real-time PPP positioning strategy which estimates two clock biases of smartphone was applied. The results show that using multi-GNSS systems data can effectively improve positioning performance; the average horizontal and vertical RMS positioning error are 0.81 and 1.65 m respectively (using GPS, BDS, and Galileo data); and the time required for each time period positioning errors in N and E directions to be under 1 m is less than 30s.


Testing the role of the cell-surface molecule Thy-1 in regeneration and plasticity of connectivity in the CNS.
Thy-1 is a cell-surface signaling molecule of the Ig superfamily implicated in the regulation of neurite outgrowth, synaptic function and plasticity. There is, however, no consensus as to its precise function in the nervous system, and it remains unclear or untested as to what its role is in the development, maintenance and plasticity of neuronal connectivity in the intact brain and whether it is essential for any of the purported functions which have been attributed to it based largely on in vitro bioassays. Here, we have engineered transgenic mice with a targeted deletion of the Thy-1 gene and, after characterizing the development of their corticospinal and thalamocortical pathways, subjected them at adulthood to paradigms of axonal regeneration and plasticity which can be readily induced during development. Quantitative analyses of the brains and spinal cords of adult null mutants showed normal cellular organization, normal anatomical features of the corticospinal and thalamocortical pathways, and basic neurophysiological properties of thalamocortical synaptic transmission which were quantitatively indistinguishable from wild-type mice. Despite the absence of Thy-1, corticospinal axons in adult mutants failed to exhibit overt regeneration following spinal cord lesion; likewise, the terminal arbors of ventrobasal thalamocortical axons also failed to reorganize in adult barrel cortex in response to whisker cautery, although they did so during a developmental critical period identical to that displayed by wild-type mice.Taken together, these results suggest that Thy-1 is not essential for the normal development and maintenance of major axon pathways and functional synaptic connections, nor would it appear to be critically important for inhibiting or promoting axonal growth, regeneration and plasticity in the developing and mature CNS.


Improvement of granulomatous skin conditions with tofacitinib in three patients: A case report.
Granulomatous skin conditions are poorly understood inflammatory skin diseases consisting predominantly of macrophages. Granuloma annulare (GA) is the most common granulomatous skin disease and the generalized variant is particularly difficult to treat due to the prolonged course and lack of efficacious treatment options. Necrobiosis lipoidica (NL) is another granulomatous disorder of uncertain etiology. There is a growing body of evidence for the use of Janus kinase (JAK) inhibitors in the management of inflammatory skin diseases. In our report, we describe three patients with recalcitrant granulomatous disease including NL and generalized GA who responded favourably to treatment with the JAK inhibitor tofacitinib. JAK inhibitors may be a beneficial therapeutic option for patients with granulomatous skin diseases that are unresponsive to conventional therapies. Further research is required to determine the long-term efficacy and safety of JAK inhibitors in treating granulomatous skin conditions.


Pre-exposure to Fe2O3 or TiO2 Nanoparticles Inhibits Subsequent Biological Uptake of 55Fe-Labeled Fe2O3 Nanoparticles.
Aquatic organisms are frequently exposed to various nanoparticles (NPs) in the natural environment. Thus, studies of NP bioaccumulation should include organisms that have been previously exposed to NPs. Our study investigated the effects of pre-exposure of Tetrahymena thermophila (T. thermophila) to Fe2O3 or TiO2 NPs on the protozoan's subsequent uptake of 55Fe-labeled Fe2O3 (55Fe2O3) NPs. Molecular mechanisms underlying the pre-exposure effects were explored in transcriptomic and metabolomic experiments. Pre-exposure to either NPs inhibited the subsequent uptake of 55Fe2O3 NPs. The results of the transcriptomic experiment indicated that NP pre-exposure influenced the expression of genes related to phagosomes and lysosomes and physiological processes such as glutathione and lipid metabolism, which are closely associated with the endocytosis of 55Fe2O3 NPs. The differentially expressed metabolites obtained from the metabolomic experiments showed an enrichment of energy metabolism and antioxidation pathways in T. thermophila pre-exposed to NPs. Together, these results demonstrate that the pre-exposure of T. thermophila to Fe2O3 or TiO2 NPs inhibited the protozoan's subsequent uptake of 55Fe2O3 NPs, possibly by mechanisms involving the alteration of endocytosis-related organelles, the induction of oxidative stress, and a lowering of the intracellular energy supply. Thus, NP pre-exposure represents a scenario which can inform increasingly realistic estimates of NP bioaccumulation.


Structure of the human activated spliceosome in three conformational states.
During each cycle of pre-mRNA splicing, the pre-catalytic spliceosome (B complex) is converted into the activated spliceosome (Bact complex), which has a well-formed active site but cannot proceed to the branching reaction. Here, we present the cryo-EM structure of the human Bact complex in three distinct conformational states. The EM map allows atomic modeling of nearly all protein components of the U2 small nuclear ribonucleoprotein (snRNP), including three of the SF3a complex and seven of the SF3b complex. The structure of the human Bact complex contains 52 proteins, U2, U5, and U6 small nuclear RNA (snRNA), and a pre-mRNA. Three distinct conformations have been captured, representing the early, mature, and late states of the human Bact complex. These complexes differ in the orientation of the Switch loop of Prp8, the splicing factors RNF113A and NY-CO-10, and most components of the NineTeen complex (NTC) and the NTC-related complex. Analysis of these three complexes and comparison with the B and C complexes reveal an ordered flux of components in the B-to-Bact and the Bact-to-B* transitions, which ultimately prime the active site for the branching reaction.


Loss of function of NCOR1 and NCOR2 impairs memory through a novel GABAergic hypothalamus-CA3 projection.
Nuclear receptor corepressor 1 (NCOR1) and NCOR2 (also known as SMRT) regulate gene expression by activating histone deacetylase 3 through their deacetylase activation domain (DAD). We show that mice with DAD knock-in mutations have memory deficits, reduced anxiety levels, and reduced social interactions. Mice with NCOR1 and NORC2 depletion specifically in GABAergic neurons (NS-V mice) recapitulated the memory deficits and had reduced GABAA receptor subunit α2 (GABRA2) expression in lateral hypothalamus GABAergic (LHGABA) neurons. This was associated with LHGABA neuron hyperexcitability and impaired hippocampal long-term potentiation, through a monosynaptic LHGABA to CA3GABA projection. Optogenetic activation of this projection caused memory deficits, whereas targeted manipulation of LHGABA or CA3GABA neuron activity reversed memory deficits in NS-V mice. We describe de novo variants in NCOR1, NCOR2 or HDAC3 in patients with intellectual disability or neurodevelopmental disorders. These findings identify a hypothalamus-hippocampus projection that may link endocrine signals with synaptic plasticity through NCOR-mediated regulation of GABA signaling.


Sequencing-based typing of HLA-B*51 alleles and the significant association of HLA-B*5101 and -B*5108 with Behçet's disease in Greek patients.
Behçet's disease (BD) is widely known to be strongly associated with human leukocyte antigen (HLA) B51 in many different ethnic groups.Recently, HLA-B51 allele typing of Greek BD patients was performed to study the distribution of B*5101-B*5107 alleles in this Greek population, the B51 antigen strongly associated with BD was found to be predominantly encoded by allele B*5101. As it is now known that the B51 antigen can be encoded by 21 alleles, B*5101-B*5121, we performed HLA-B*51 allele genotyping among 58 Greek patients with BD. After serological HLA typing, typing of HLA-B*51 alleles was performed using the polymerase chain reaction-sequencing-based typing (PCR-SBT) method. The frequency of the B51 antigen was found to be significantly higher in the patient group as compared with the control group (75.9% of patients vs 22.0% of controls. In the genotyping of B51 alleles, 34 out of 44 B51-positive patients possessed B*5101, 13 out of the 44 carried B*5108. In contrast, all of the 9 B51-positive normal controls carried B*5101. This study revealed a strong association between Greeks with BD, both B*5101, B*5108, provided important insights into the molecular mechanism underlying the association between HLA status, this disease.


[Sensitivity of Salmonella species to four "classical" antibiotics and seven new beta-lactamic antibiotics (author's transl)].
A study of the minimal inhibitory concentration (MIC) for 200 strains of Salmonella sp. has been performed with the following beta-lactamic antibiotics: mezlocillin and azlocillin (penicillins), cephaclor, cephamandole, cephuroxime and cephotaxime (cephalosporins), and cephoxytine (cephamycin). The MIC has been compared with that of chloramphenicol, ampicillin, amoxicillin and cotrimoxazole, all of them widely used antibiotics for the treatment of Salmonella infections in Spain. The different species and serotypes of Salmonella studied were all sensitive to all the beta-lactamic antibiotics tested. Of particular relevance is the fact that cephotaxime (HR 756) MIC was extraordinarily low even for strains resistent to the penicillins. The MIC of cephaclor, and oral cephalosporin, was similar to that of the parenteral cephalosporins.


Prediction of hepatic plasma clearance in vivo from gel-entrapped rat and human hepatocytes.
This paper aimed to evaluate the applicability of gel-entrapped rat and human hepatocytes in the prediction of hepatic plasma clearance (CLh,plasma) in vivo. The in vitro intrinsic clearances (CLint,in vitro) for the selected compounds were determined from the substrate disappearance rate, and further used to predict CLh,plasma using 3 classical mathematical models (well-stirred, parallel-tube, and dispersion) and disregarding drug binding. As a result, the predicted values from gel-entrapped rat hepatocytes were mostly within 2 SE of the literature data with a high correlation coefficient (R(2)) of 0.88-0.91. The predicted data with human hepatocytes also fitted well with the clinical data, indicating a high accuracy in prediction of in-vivo clearance. With respect to the mathematical model for predicting CLh,plasma, the parallel-tube and dispersion models produced a better prediction than the well-stirred model, and we suggest using the parallel-tube model because it is less complex mathematically. In conclusion, gel-entrapped hepatocytes predicted the drug clearance well and seemed to be a useful tool in the process of drug discovery.


Association Between Preoperative Factors and In-hospital Mortality in Neonates After Cardiac Surgery in China.
Background: Little is known about preoperative factors affecting cardiac surgery outcomes of neonates in China. We sought to examine the association between characteristics of neonates with congenital heart disease (CHD) and early postoperative outcomes after cardiac repair in a tertiary care paediatric hospital. Methods: A single-centre retrospective cohort study of neonates who underwent cardiac surgery between January 2006 and December 2019 was performed. Demographic, institutional, and surgical characteristics of neonates were examined and their association with in-hospital mortality was analysed using multivariable logistic regression models. Results: During the study period, we analysed the outcomes of 1,078 neonates. In-hospital mortality decreased to 13.8% in the era 2017-2019. The overall in-hospital mortality rate was 16.3%. Normal weight at surgery [odds ratio (OR), 0.63; 95% confidence interval (CI), 0.47-0.85; P = 0.003] was associated with lower mortality risk. Poor health status (emergent: OR, 3.11; 95% CI, 1.96-4.94; P < 0.001; elective: OR, 1.63; 95% CI, 1.11-2.40; P = 0.013), higher Society of Thoracic Surgeons-European Association for Cardio-Thoracic Surgery (STAT) categories (STAT 5 category: OR, 2.58; 95% CI, 1.04-6.43; P = 0.042), and limited individual surgeon experience (surgeon with 5-10 operations per year: OR, 1.43; 95% CI, 1.06-1.95; P = 0.021) were associated with higher odds of early death. Conclusion: In-hospital mortality after neonatal cardiac surgery remained high in our centre over the past 10 years. Some preoperative aspects, including low-weight at surgery, poor health status, increased surgical complexity, and limited surgeon experience were significantly associated with higher mortality. Based on the observed associations, the necessary practises to be modified, especially in preoperative care, should be identified and assessed in future research.


Initial experience and sizing considerations using the three lobed amplatzer ductal occluder.
The recently released Amplatzer Ductal Occluder 2 (ADO2) was designed specifically for use in small children with moderate-sized shunts and larger children with small patent ductus arterioses (PDA). We report our initial experience with this device. Patients referred with PDA underwent occlusion using the ADO2. This is a fabric-free nitinol wire 3-lobed device. All cases underwent pre-, intra- and post-procedural echocardiography, with follow up at 1 day and one month. Device sizing for device waist diameter and width was based on aortography. Seven patients with a median age of 3 years and 4 months (range 7 months-23 years) and a median weight of 12 kg (range 7-56 kg) underwent successful PDA closure. The median ductal diameter was 1.5 mm (range 0.4-4 mm). Both transpulmonary (6/7) and transaortic approaches (1/7) were used. Recurrent patency at 24 hours with complete occlusion at 1 month was noted in a single case. A specific device-based length assessment applied resulted in shorter than recommended device selection. The ADO2 broadens the spectrum of PDAs that can be simply and safely treated with devices. The flexibility of the articulations, coupled with the alternative deployment techniques, allow for increased ease of treatment in a range of small patients and specific ductal anatomies. An alternative device-specific length measurement of the duct length may result in less retaining disc protrusion. Broader experience is required to further delineate device and patient selection as well as to document its long-term efficacy and safety.


The action of phorbol ester on steroidogenesis in cultured human fetal adrenal cells.
The potent mitogen and tumor promoter, phorbol 12-myristate 13-acetate (PMA), has a primary action via activation of calcium-dependent protein kinase C. The treatment of monolayer cultures of human fetal adrenal neocortex (HFA) cells with PMA (50-250 nM) stimulated basal dehydroepiandrosterone sulfate (DS) secretion 2-3 fold. ACTH-treated HFA cells secreted amounts of DS and cortisol (F) 10-50 fold greater than basal secretions. PMA (250 nM) addition with ACTH to HFA cells decreased DS and F secretions at least 75% on days 2 and 3 of treatment. Treatment of HFA cells with 4 alpha-phorbol, which does not activate calcium-dependent protein kinase C, did not inhibit steroidogenesis. The attenuated rates of steroidogenesis after PMA treatment correlated with the decreased amounts of steroid 11 beta, 17 alpha- and 21-hydroxylase cholesterol side-chain cleavage steroid dehydrogenase and sulfotransferase activities. The decrease of steroid 17 alpha-hydroxylase activity correlated with the decreased amount of cytochrome P-450(17) alpha as determined after protein immunoblotting of NaDodSO4 cell lysates. After PMA treatment the ACTH-promoted increases of hydroxysteroid sulfotransferase and dehydrogenase activities of HFA cells were suppressed. PMA (50 nM) inhibited cAMP accumulation in ACTH-treated HFA cells, while 4 alpha-phorbol had no effect. Importantly, dibutyryl cAMP (0.2 mM) treatment of HFA cells did not reverse phorbol ester-promoted attenuation of steroidogenesis. We conclude that, in the presence of ACTH, phorbol ester chronically inhibits both cAMP synthesis and cAMP-dependent protein kinase action with resultant decreased steroidogenic enzyme synthesis and steroid production. This may be a consequence of activation, migration and a slow degradation of protein kinase C activity. These multifaceted actions of phorbol ester and associated protein kinase C activation may have critical effects on the ontogeny of fetal adrenal function.


Effects of laparoscopic surgery on serum anti-Müllerian hormone levels in reproductive-aged women with endometrioma.
A few studies have investigated the effects of laparoscopic cystectomy for endometrioma on ovarian reserve using serum anti-Müllerian hormone (AMH) levels and have shown inconsistent results. This prospective study was performed to estimate early changes in serum AMH after laparoscopic surgery for endometrioma. In 27 women aged 21-46 years who underwent laparoscopic unilateral cystectomy (n = 13) or oophorectomy (n = 14) for endometrioma, we measured pre- and postoperative serum AMH levels at baseline and 1 week, 1 month, and 3 months after surgery. Serial changes in serum AMH levels were analyzed according to the operation type using analysis of variance. Mean levels of serum AMH decreased significantly immediately after surgery and remained for up to 3 months in both operation types (P = 0.002 for cystectomy group and P <0.001 for oophorectomy group). In addition, the serum AMH levels showed similar patterns of change after cystectomy or oophorectomy. In conclusion, laparoscopic ovarian cystectomy for endometrioma appears to decrease serum AMH levels immediately after surgery.


Relatively increased von Willebrand factor activity after off-pump coronary artery bypass graft surgery.
High shear forces can induce structural changes in the shape of the von Willebrand factor (VWF) molecule, making it more sensitive for ADAMTS-13, a specific VWF protease. This leads to proteolysis of the highest-molecular-weight multimers, which are the most effective in platelet-mediated hemostasis under conditions of high shear stress. The use of cardiopulmonary bypass (CPB) is accompanied with high shear forces and could therefore lead to diminished VWF activity. Therefore, we studied VWF activity in patients undergoing myocardial revascularization. We enrolled 60 patients undergoing coronary artery bypass graft (CABG) surgery with and without CPB (30 in each group). ADAMTS-13 activity, VWF antigen (VWF:Ag) and propeptide levels were measured directly before and after the procedure. VWF activity was determined using both the ristocetin cofactor activity (VWF:Rcof) and collagen binding (VWF:CB) assays. VWF:Rcof and VWF:CB, both corrected for VWF:Ag, were significantly increased after the procedure in the off-pump group (p < 0.001 and p = 0.05, respectively), but not in the CPB group. Postoperative VWF:Ag and VWF-propeptide levels significantly increased in both groups. ADAMTS-13 activity increased after both types of surgery after correction for hemodilution. Postoperative VWF:Rcof levels correlated with postoperative D-dimer levels (p = 0.025) and were associated with early cognitive decline in the off-pump group (p = 0.025). Our data indicate that VWF activity is increased after off-pump CABG, but not after CABG with CPB. Since release of VWF is comparable in both groups, shear stress-induced proteolysis due to the use of CPB may be responsible for the decreased VWF activity.


Microbiological oxidation of antimony(III) with oxygen or nitrate by bacteria isolated from contaminated mine sediments.
Bacterial oxidation of arsenite [As(III)] is a well-studied and important biogeochemical pathway that directly influences the mobility and toxicity of arsenic in the environment. In contrast, little is known about microbiological oxidation of the chemically similar anion antimonite [Sb(III)]. In this study, two bacterial strains, designated IDSBO-1 and IDSBO-4, which grow on tartrate compounds and oxidize Sb(III) using either oxygen or nitrate, respectively, as a terminal electron acceptor, were isolated from contaminated mine sediments. Both isolates belonged to the Comamonadaceae family and were 99% similar to previously described species. We identify these novel strains as Hydrogenophaga taeniospiralis strain IDSBO-1 and Variovorax paradoxus strain IDSBO-4. Both strains possess a gene with homology to the aioA gene, which encodes an As(III)-oxidase, and both oxidize As(III) aerobically, but only IDSBO-4 oxidized Sb(III) in the presence of air, while strain IDSBO-1 could achieve this via nitrate respiration. Our results suggest that expression of aioA is not induced by Sb(III) but may be involved in Sb(III) oxidation along with an Sb(III)-specific pathway. Phylogenetic analysis of proteins encoded by the aioA genes revealed a close sequence similarity (90%) among the two isolates and other known As(III)-oxidizing bacteria, particularly Acidovorax sp. strain NO1. Both isolates were capable of chemolithoautotrophic growth using As(III) as a primary electron donor, and strain IDSBO-4 exhibited incorporation of radiolabeled [(14)C]bicarbonate while oxidizing Sb(III) from Sb(III)-tartrate, suggesting possible Sb(III)-dependent autotrophy. Enrichment cultures produced the Sb(V) oxide mineral mopungite and lesser amounts of Sb(III)-bearing senarmontite as precipitates.


Itaconimides as Novel Quorum Sensing Inhibitors of Pseudomonas aeruginosa.
Pseudomonas aeruginosa is known as an opportunistic pathogen that often causes persistent infections associated with high level of antibiotic-resistance and biofilms formation. Chemical interference with bacterial cell-to-cell communication, termed quorum sensing (QS), has been recognized as an attractive approach to control infections and address the drug resistance problems currently observed worldwide. Instead of imposing direct selective pressure on bacterial growth, the right bioactive compounds can preferentially block QS-based communication and attenuate cascades of bacterial gene expression and production of virulence factors, thus leading to reduced pathogenicity. Herein, we report on the potential of itaconimides as quorum sensing inhibitors (QSI) of P. aeruginosa. An initial hit was discovered in a screening program of an in-house compound collection, and subsequent structure-activity relationship (SAR) studies provided analogs that could reduce expression of central QS-regulated virulence factors (elastase, rhamnolipid, and pyocyanin), and also successfully lead to the eradication of P. aeruginosa biofilms in combination with tobramycin. Further studies on the cytotoxicity of compounds using murine macrophages indicated no toxicity at common working concentrations, thereby pointing to the potential of these small molecules as promising entities for antimicrobial drug development.


Calculated and measured [Ca(2+)] in buffers used to calibrate Ca(2+) macroelectrodes.
The ionized concentration of calcium in physiological buffers ([Ca(2+)]) is normally calculated using either tabulated constants or software programs. To investigate the accuracy of such calculations, the [Ca(2+)] in EGTA [ethylene glycol-bis(β-aminoethylether)-N,N,N|,N|-tetraacetic acid], BAPTA [1,2-bis(o-aminophenoxy) ethane-N,N,N|,N|-tetraacetic acid], HEDTA [N-(2-hydroxyethyl)-ethylenediamine-N,N|,N|-triacetic acid], and NTA [N,N-bis(carboxymethyl)glycine] buffers was estimated using the ligand optimization method, and these measured values were compared with calculated values. All measurements overlapped in the pCa range of 3.51 (NTA) to 8.12 (EGTA). In all four buffer solutions, there was no correlation between measured and calculated values; the calculated values differed among themselves by factors varying from 1.3 (NTA) to 6.9 (EGTA). Independent measurements of EGTA purity and the apparent dissociation constants for HEDTA and NTA were not significantly different from the values estimated by the ligand optimization method, further substantiating the method. Using two calibration solutions of pCa 2.0 and 3.01 and seven buffers in the pCa range of 4.0-7.5, calibration of a Ca(2+) electrode over the pCa range of 2.0-7.5 became a routine procedure. It is proposed that such Ca(2+) calibration/buffer solutions be internationally defined and made commercially available to allow the precise measurement of [Ca(2+)] in biology.


The adverse effects of comorbid pain on depression outcomes in primary care patients: results from the ARTIST trial.
To explore the effect of pain symptoms and improvements in pain on depression outcomes. We analyzed data from A Randomized Trial Investigating SSRI Treatment (ARTIST), a randomized longitudinal effectiveness study comparing selective serotonin reuptake inhibitors (SSRIs) for the treatment of depression in primary care (n = 573). Depression outcome at month 6, defined as remission, partial response, and nonresponse using the Symptom Checklist-20, was the primary outcome. Compared to patients with no pain at baseline, those with severe pain were less likely to achieve remission (OR = 0.11, 95% CI 0.05-0.25) and partial response (OR = 0.24, 95% CI 0.10-0.59) vs nonresponse. Patients with moderate pain were less likely to achieve remission vs nonresponse (OR = 0.25, 95% CI 0.13-0.48). Patients with early improvement in pain were more likely to achieve remission (OR = 1.90, 95% CI 1.03-3.49). Accounting for missing data with last observation carried forward or multiple imputation yielded similar results. Pain symptoms are present in the majority of depressed primary care patients beginning antidepressant therapy. Pain symptoms are associated with worse depression outcomes, while improvement in pain is associated with significantly better depression outcomes. Attention to comorbid pain may be important in enhancing depression care.


Clinical utility of computed tomography in the diagnosis of pulmonary embolism.
This prospective clinical pilot study describes the clinical utility and cost effectiveness of computed tomography (CT) with contrast in the diagnosis and management of pulmonary embolism. The setting is a university teaching hospital, and the 20 patients, 26 to 81 years old, were found to have CT findings consistent with pulmonary embolism. Intraluminal pulmonary artery clots were observed on CT and contributed to clinical management, often obviating pulmonary arteriography. CT, particularly spiral CT, may demonstrate pulmonary embolism and offers advantages over ventilation-perfusion lung scanning and pulmonary arteriography in making the diagnosis of pulmonary embolism in high-risk patients or patients with preexisting parenchymal lung disease.


Lung Transplantation for Patients With COVID-19.
The COVID-19 pandemic has caused acute lung injury in millions of individuals worldwide. Some patients develop COVID-related acute respiratory distress syndrome (CARDS) and cannot be liberated from mechanical ventilation. Others may develop post-COVID fibrosis, resulting in substantial disability and need for long-term supplemental oxygen. In both of these situations, treatment teams often inquire about the possibility of lung transplantation. In fact, lung transplantation has been successfully employed for both CARDS and post-COVID fibrosis in a limited number of patients worldwide. Lung transplantation after COVID infection presents a number of unique challenges that transplant programs must consider. In those with severe CARDS, the inability to conduct proper psychosocial evaluation and pretransplantation education, marked deconditioning from critical illness, and infectious concerns regarding viral reactivation are major hurdles. In those with post-COVID fibrosis, our limited knowledge about the natural history of recovery after COVID-19 infection is problematic. Increased knowledge of the likelihood and degree of recovery after COVID-19 acute lung injury is essential for appropriate decision-making with regard to transplantation. Transplant physicians must weigh the risks and benefits of lung transplantation differently in a post-COVID fibrosis patient who is likely to remain stable or gradually improve in comparison with a patient with a known progressive fibrosing interstitial lung disease (fILD). Clearly lung transplantation can be a life-saving therapeutic option for some patients with severe lung injury from COVID-19 infection. In this review, we discuss how lung transplant providers from a number of experienced centers approach lung transplantation for CARDS or post-COVID fibrosis.


Pretreatment total lymphocyte count as a prognostic factor of survival in patients with recurrent cervical cancer after definitive radiation-based therapy: a retrospective study.
This study evaluated the association between pretreatment total lymphocyte count (TLC) and overall survival (OS) in patients with recurrent cervical cancer. We retrospectively reviewed 290 patients with recurrent cervical cancer with definite complete responses to either definitive radiotherapy or concurrent chemoradiotherapy between January 2009 and December 2022. The associations between pretreatment TLC and progression-free survival (PFS) and OS rates were evaluated. Ninety-three patients (32%) had a pretreatment TLC <1,000 cells/mm3. Patients with a pretreatment TLC <1,000 cells/mm3 had lower treatment response rates than their counterparts (P=0.045). The OS and PFS rates were significantly higher in patients with pretreatment TLC ≥1,000 cells/mm3 than in those with pretreatment TLC <1,000 cells/mm3 (10.74 vs. 3.89 months, P<0.0001; 8.32 vs. 4.97 months, P=0.042; respectively). Moreover, pretreatment TLC ≥1,000 cells/mm3 was identified as an independent prognostic factor for OS in both univariate analysis (hazard ratio [HR], 0.57; 95% conficence interval [CI], 0.44-0.74; P<0.001) and multivariate analysis (HR, 0.64; 95% CI, 0.47-0.86; P=0.003). However, TLC ≥1,000 cells/mm3 was identified as a prognostic factor for PFS only in univariate analysis (HR, 0.71; 95% CI, 0.51-0.99; P=0.043) but not in the multivariate analysis (HR, 0.81; 95% CI, 0.55-1.18; P=0.3). Pretreatment TLC was associated with treatment response and was identified as an independent prognostic factor associated with the survival outcomes of patients with recurrent cervical cancer.


Giardia duodenalis genotypes among schoolchildren and their families and pets in urban and rural areas of Sinaloa, Mexico.
Giardiasis is a human health concern worldwide, especially among schoolchildren. Giardia duodenalis genotypes A and B are infective to humans, but their zoonotic potential remains controversial. In Mexico, the most prevalent genotype is A, but B was also detected in southeastern Mexico. In Sinaloa state, northwestern Mexico, giardiasis is highly prevalent, but Giardia genotypes have been poorly studied. This study aimed to investigate the distribution and clinical-epidemiological correlation of G. duodenalis genotypes in schoolchildren and their families and pets in urban and rural areas of Sinaloa state, Mexico. Among 395 schoolchildren (274 urban, 121 rural), 76 (49 urban, 27 rural) were infected with G. duodenalis. In total, 22 families (15 urban, 7 rural) of infected schoolchildren, consisting of 60 family members (41 urban, 19 rural) and 21 pet dogs (15 urban, 6 rural) were examined; 10 family members (5 urban, 5 rural) and 5 pet dogs (3 urban, 2 rural) of 10 families (6 urban, 4 rural) were infected. After PCR-RFLP analyses of vsp417 and gdh genes, genotype prevalence among infected urban schoolchildren was 79.5% AI, 12.8% AII, and 7.7% mixed AI+B. However, only AI genotype was found in family members and pets. In the rural area, only the AI genotype was detected. Genotypes were not correlated with clinical manifestations. This paper shows the presence of B genotype in northwestern Mexico for the first time. Detection of AI genotype in dogs suggested the possible role of dogs as the reservoir for human giardiasis in Sinaloa, Mexico.


In vivo functional characterization of the aldolase B gene enhancer.
A 400-bp intronic enhancer fragment in conjunction with the proximal promoter of the aldolase B gene provided correct tissue-specific expression in transgenic mice together with hormonal regulation in the liver. We investigated in vivo and in cultured cells the contribution of the intronic regulatory sequences and their interaction with the promoter elements in controlling aldolase B gene expression. Transgene activity was completely abolished by disruption of the two hepatocyte nuclear factor 1 (HNF1) binding sites in the enhancer, whereas mutation of one HNF1 site had no effect in the liver but strongly decreased activity in the kidney. Our data show that the HNF1 binding site(s) in the enhancer were key regulators of aldolase B transgene expression both in the liver and kidney. Deletion of the CCAAT/enhancer-binding protein site in the promoter completely abolished the enhancer function in HepG2 cells. These results suggest that expression of the aldolase B gene in the liver requires cooperative interactions between CCAAT/enhancer-binding protein and HNF1. Deletion of the HNF4 binding site in the enhancer suppressed expression in both liver and kidney in half of the transgenic lines, suggesting that this element might play a role in chromatin opening at the insertion site. We firmly establish that the endogenous aldolase B gene's first response to glucagon or cyclic AMP exposure was a transient increase in the expression in the liver, followed by a secondary decline in the transcription, as previously reported. This response was reproduced by all transgenes studied, indicating that neither HNF1 nor HNF4 binding sites in the enhancer were involved in this biphasic cyclic AMP response.


[Variability of the deep femoral venous system].
Deep venous system is known for its extreme variability but in anatomy it receives only marginal interest. Although a few previous anatomical studies have already pointed out the fact of a significant discrepancy between the autopsy findings and the literary description, it has not had any particular output so far. Our findings confirmed the deep femoral vein to be an alternative collateral vein connecting the popliteal with the femoral vein.


Leaf Treatments with a Protein-Based Resistance Inducer Partially Modify Phyllosphere Microbial Communities of Grapevine.
Protein derivatives and carbohydrates can stimulate plant growth, increase stress tolerance, and activate plant defense mechanisms. However, these molecules can also act as a nutritional substrate for microbial communities living on the plant phyllosphere and possibly affect their biocontrol activity against pathogens. We investigated the mechanisms of action of a protein derivative (nutrient broth, NB) against grapevine downy mildew, specifically focusing on the effects of foliar treatments on plant defense stimulation and on the composition and biocontrol features of the phyllosphere microbial populations. NB reduced downy mildew symptoms and induced the expression of defense-related genes in greenhouse- and in vitro-grown plants, indicating the activation of grapevine resistance mechanisms. Furthermore, NB increased the number of culturable phyllosphere bacteria and altered the composition of bacterial and fungal populations on leaves of greenhouse-grown plants. Although, NB-induced changes on microbial populations were affected by the structure of indigenous communities originally residing on grapevine leaves, degrees of disease reduction and defense gene modulation were consistent among the experiments. Thus, modifications in the structure of phyllosphere populations caused by NB application could partially contribute to downy mildew control by competition for space or other biocontrol strategies. Particularly, changes in the abundance of phyllosphere microorganisms may provide a contribution to resistance induction, partially affecting the hormone-mediated signaling pathways involved. Modifying phyllosphere populations by increasing natural biocontrol agents with the application of selected nutritional factors can open new opportunities in terms of sustainable plant protection strategies.


In situ silver nanoparticle development for molecular-specific biological imaging via highly accessible microscopies.
In biological studies and diagnoses, brightfield (BF), fluorescence, and electron microscopy (EM) are used to image biomolecules inside cells. When compared, their relative advantages and disadvantages are obvious. BF microscopy is the most accessible of the three, but its resolution is limited to a few microns. EM provides a nanoscale resolution, but sample preparation is time-consuming. In this study, we present a new imaging technique, which we termed decoration microscopy (DecoM), and quantitative investigations to address the aforementioned issues in EM and BF microscopy. For molecular-specific EM imaging, DecoM labels proteins inside cells using antibodies bearing 1.4 nm gold nanoparticles (AuNPs) and grows silver layers on the AuNPs' surfaces. The cells are then dried without buffer exchange and imaged using scanning electron microscopy (SEM). Structures labeled with silver-grown AuNPs are clearly visible on SEM, even they are covered with lipid membranes. Using stochastic optical reconstruction microscopy, we show that the drying process causes negligible distortion of structures and that less structural deformation could be achieved through simple buffer exchange to hexamethyldisilazane. Using DecoM, we visualize the nanoscale alterations in microtubules by microtubule-severing proteins that cannot be observed with diffraction-limited fluorescence microscopy. We then combine DecoM with expansion microscopy to enable sub-micron resolution BF microscopy imaging. We first show that silver-grown AuNPs strongly absorb white light, and the structures labeled with them are clearly visible on BF microscopy. We then show that the application of AuNPs and silver development must follow expansion to visualize the labeled proteins clearly with sub-micron resolution.


Phototransposition reactions of arylboronate esters in acetonitrile and 2,2,2-trifluoroethanol.
The phototransposition (para, meta, ortho) reactions of the arylboronate esters 4-, 3-, and 2-(4',4',5',5'-tetramethyl-1',3',2'-dioxaborolanyl)toluenes (1, 2, and 3, respectively) in both acetonitrile and 2,2,2-trifluoroethanol (TFE) using 254 nm irradiation have been examined. The irradiations resulted in steady-state compositions of para (5%), meta (19%), and ortho (76%) isomers in acetonitrile starting from the ortho isomer and para (12%), meta (54%), and ortho (35%) isomers in TFE starting from the para isomer. Analysis of the (13)C NMR spectrum of the product mixture obtained from the photochemistry of the para isomer selectively deuterated at C3 and C5 (1d(2)()) revealed that the boron-substituted carbon is the active one in the phototransposition reactions in both acetonitrile and TFE. Similar results were observed for irradiations of 1 in cyclohexane. Fluorescence spectra, singlet-state lifetimes, and Stern-Volmer quenching of fluorescence with 2,3-dimethyl-1,3-butadiene indicated that the excited singlet states of these three isomers were spectroscopic minima and that the excited singlet state was the reactive one for 3 in acetonitrile.


Aggregation of chemotactic organisms in a differential flow.
We study the effect of advection on the aggregation and pattern formation in chemotactic systems described by Keller-Segel-type models. The evolution of small perturbations is studied analytically in the linear regime complemented by numerical simulations. We show that a uniform differential flow can significantly alter the spatial structure and dynamics of the chemotactic system. The flow leads to the formation of anisotropic aggregates that move following the direction of the flow, even when the chemotactic organisms are not directly advected by the flow. Sufficiently strong advection can stop the aggregation and coarsening process that is then restricted to the direction perpendicular to the flow.