Pleiotropic resistance to DNA-interactive drugs is associated with increased expression of genes involved in DNA replication, repair, and stress response.
A combination of four genetic suppressor elements (GSEs), two of which are derived from putative transcriptional regulators, was previously found to increase resistance to drugs inhibiting DNA replication in HT1080 fibrosarcoma cells. In the present study, two GSE-transduced cell lines, isolated with and without cytotoxic selection, were found to be resistant to a diverse group of DNA-interactive agents, including aphidicolin, hydroxyurea, cytarabine, etoposide, doxorubicin, and mafosfamide. Changes in gene expression associated with GSE-induced drug resistance were analyzed by cDNA array hybridization and reverse transcription-PCR. Twenty genes were found to be up-regulated in both of the resistant cell lines. These include genes involved in DNA replication and repair (e.g., PCNA, XRCC1, B-MYB, and GADD45), transcriptional regulators associated with stress response, and cell cycle checkpoint control (e.g., YB-1, DBPA, and ATF4), and genes for signal transduction proteins (e.g., protein tyrosine phosphatase 1B and regulatory subunits alpha and beta of cAMP-dependent protein kinase). The observed changes in gene expression may play a role in pleiotropic resistance to different classes of DNA-targeting drugs.


Ivermectin also inhibits the replication of bovine respiratory viruses (BRSV, BPIV-3, BoHV-1, BCoV and BVDV) in vitro.
Bovine respiratory disease (BRD) complex is an important viral infection that causes huge economic losses in cattle herds worldwide. However, there is no directly effective antiviral drug application against respiratory viral pathogens; generally, the metaphylactic antibacterial drug applications are used for BRD. Ivermectin (IVM) is currently used as a broad-spectrum anti-parasitic agent both for veterinary and human medicine on some occasions. Moreover, since it is identified as an inhibitor for importin α/β-mediated nuclear localization signal (NLS), IVM is also reported to have antiviral potential against several RNA and DNA viruses. Since therapeutic use of IVM in COVID-19 cases has recently been postulated, the potential antiviral activity of IVM against bovine respiratory viruses including BRSV, BPIV-3, BoHV-1, BCoV and BVDV are evaluated in this study. For these purposes, virus titration assay was used to evaluate titers in viral harvest from infected cells treated with non-cytotoxic IVM concentrations (1, 2.5 and 5 μM) and compared to titers from non-treated infected cells. This study indicated that IVM inhibits the replication of BCoV, BVDV, BRSV, BPIV-3 and BoHV-1 in a dose-dependent manner in vitro as well as number of extracellular infectious virions. In addition, it was demonstrated that IVM has no clear effect on the attachment and penetration steps of the replication of the studied viruses. Finally, this study shows for the first time that IVM can inhibit infection of BRD-related viral agents namely BCoV, BPIV-3, BVDV, BRSV and BoHV-1 at the concentrations of 2.5 and 5 μM. Consequently, IVM, which is licensed for antiparasitic indications, also deserves to be evaluated as a broad-spectrum antiviral in BRD cases caused by viral pathogens.


Enzymatic Oxy- and Amino-Functionalization in Biocatalytic Cascade Synthesis: Recent Advances and Future Perspectives.
Biocatalytic cascades are a powerful tool for building complex molecules containing oxygen and nitrogen functionalities. Moreover, the combination of multiple enzymes in one pot offers the possibility to minimize downstream processing and waste production. In this review, we illustrate various recent efforts in the development of multi-step syntheses involving C-O and C-N bond-forming enzymes to produce high value-added compounds, such as pharmaceuticals and polymer precursors. Both in vitro and in vivo examples are discussed, revealing the respective advantages and drawbacks. The use of engineered enzymes to boost the cascades outcome is also addressed and current co-substrate and cofactor recycling strategies are presented, highlighting the importance of atom economy. Finally, tools to overcome current challenges for multi-enzymatic oxy- and amino-functionalization reactions are discussed, including flow systems with immobilized biocatalysts and cascades in confined nanomaterials.


Molecular genetics and antisocial behavior: where do we stand?
Over the last two decades, it has become increasingly evident that control of aggressive behavior is modulated by the individual genetic profile as well. Several candidate genes have been proposed to play a role in the risk to develop antisocial behavior, and distinct brain imaging studies have shown that specific cortical areas may be functionally and/or structurally impaired in impulsive violent subjects on the basis of their genotypes. In this paper, we review the findings regarding four polymorphisms-MAOA (Monoamine oxidase A) uVNTR, SLC6A4 (solute carrier family 6 (neurotransmitter transporter), member 4) 5HTTLPR, COMT (Catechol-O-methyltransferase) Val158Met and DRD4 (dopamine D4 receptor) VNTR 1-11-that all have been found to be associated with an increased vulnerability for antisocial and impulsive behavior in response to aversive environmental conditions. These results, however, have not been replicated by other studies, likely because of crucial methodological discrepancies, including variability in the criteria used to define antisocial behavior and assessment of environmental factors. Finally, it has been recently proposed that these genetic variants may actually increase the individual susceptibility not merely to the negative environmental factors, but to the positive ones as well. In this view, such alleles would play a wider modulatory role, by acting as "plasticity" rather than "vulnerability" genes. Overall, these findings have potential important implications that span well outside of neuroscience and psychiatry, to embrace ethics, philosophy, and the law itself, as they pose new challenges to the very notion of Free Will. Novel properly controlled studies that examine multi-allelic genetic profiles, rather than focusing on distinct single variants, will make it possible to achieve a clearer understanding of the molecular underpinnings of the nature by nurture interaction.


Predictors of adverse outcomes in HIV-1-infected children receiving combination antiretroviral treatment: results from a DREAM cohort in sub-Saharan Africa.
HIV-infected children have less access to combination antiretroviral therapy as compared with adults in resource-limited settings. Growth faltering, loss to follow-up (LTFU) and high mortality are frequently seen. A retrospective cohort study was performed with parameters extracted from the Drug Resource Enhancement against AIDS and Malnutrition database for HIV-infected, antiretroviral naïve children under 15 years presenting for care at 17 Drug Resource Enhancement against AIDS and Malnutrition centers in Mozambique, Malawi and Guinea between January 2005 to December 2008. Predictors of time-to-death, time-to-LTFU and persistence of malnutrition by Cox's regression and Kaplan-Meier were determined. 2215 children presented to care with 1343 (61%) being ≤ 5 years. At baseline, stunting and malnutrition occurred in 40% and 25%, respectively; 75% of 2149 children had CD4 cell percentages less than 20; median HIV RNA, log10 cp/mL, was 4.97 in 1927 patients. Over time 238 children died (10.7%; 2.7% person-years [PY]) 63 were LTFU (2.8%; 0.7% PY). By multivariate analysis, mortality was associated with virus load (hazards ratio: 1.19; confidence interval: 1.01-1.402, P = 0.038) and reduced weight-for-age Z scores (hazards ratio: 0.590; confidence interval: 0.53-0.66, P < 0.001). LTFU was associated with low weight-for-height Z scores (hazards ratio: 0.71; confidence interval: 0.51-0.97, P = 0.031). At 12 months after combination antiretroviral therapy, anthropometric parameters significantly improved in 1226 children (P < 0.001); virus load declined to <400 copies/mL in over 60%. Despite advanced HIV disease, children initiating combination antiretroviral therapy had mortality rates of 2.7% p/PY with overall attrition rates of 11.7% p/100 PY, with significant reversal of negative anthropometric markers, and improvement of immunological and virological parameters in children with 12 months of follow-up.


Transcriptomic response of cowpea bruchids to N-acetylglucosamine-specific lectins.
Griffonia simplicifolia lectin II (GSII) and wheat germ agglutinin (WGA) are N-acetylglucosamine-binding lectins. Previous studies demonstrated that they have anti-insect activity, a property potentially useful in pest control. To gain some insight into the insect response to dietary lectins, we performed transcriptomic analysis using the cowpea bruchid (Callosobruchus maculatus) midgut microarray platform we built. Compared to the nonnutritional cellulose treatment, dietary lectins induced more profound changes in gene expression. Ingestion of relatively high doses of lectins for 24 h resulted in alteration of gene expression involved in sugar and lipid metabolism, transport, development, defense, and stress tolerance. Metabolic genes were largely downregulated. Moreover, we observed disorganized microvilli resulting from ingestion of WGA. This morphological change is consistent with the lectin-induced changes in genes related to midgut epithelial cell repair. In addition, suboptimal nutrient conditions may serve as a stress signal to trigger senescence processes, leading to growth arrest and developmental delay.


What Can Computational Modeling Tell Us about the Diversity of Odor-Capture Structures in the Pancrustacea?
A major transition in the history of the Pancrustacea was the invasion of several lineages of these animals onto land. We investigated the functional performance of odor-capture organs, antennae with olfactory sensilla arrays, through the use of a computational model of advection and diffusion of odorants to olfactory sensilla while varying three parameters thought to be important to odor capture (Reynolds number, gap-width-to-sensillum-diameter ratio, and angle of the sensilla array with respect to oncoming flow). We also performed a sensitivity analysis on these parameters using uncertainty quantification to analyze their relative contributions to odor-capture performance. The results of this analysis indicate that odor capture in water and in air are fundamentally different. Odor capture in water and leakiness of the array are highly sensitive to Reynolds number and moderately sensitive to angle, whereas odor capture in air is highly sensitive to gap widths between sensilla and moderately sensitive to angle. Leakiness is not a good predictor of odor capture in air, likely due to the relative importance of diffusion to odor transport in air compared to water. We also used the sensitivity analysis to make predictions about morphological and kinematic diversity in extant groups of aquatic and terrestrial crustaceans. Aquatic crustaceans will likely exhibit denser arrays and induce flow within the arrays, whereas terrestrial crustaceans will rely on more sparse arrays with wider gaps and little-to-no animal-induced currents.


Integrin alpha(v)beta(3)-Targeted Cancer Therapy.
Anti-angiogenesis is a promising strategy for the treatment of cancer. Integrins, consisting of two noncovalently bound transmembrane alpha and beta subunits, are an important molecular family involved in tumor angiogenesis. The blockade of integrin signaling has been demonstrated to be efficient to inhibit tumor growth, angiogenesis, and metastasis. Among all the integrins, alpha(v)beta(3) seems to be the most important one during tumor angiogenesis. The inhibition of integrin alpha(v)beta(3) signaling with antibodies, peptides, peptidomimetics, and other antagonists has great potential in the treatment of cancer. In addition, integrin alpha(v)beta(3) is highly expressed on activated endothelial cells, new-born vessels as well as some tumor cells, but is not present in resting endothelial cells and most normal organ systems, making it a suitable target for anti-angiogenic therapy. In this article we will review the role of integrin alpha(v)beta(3) in angiogenesis, present recent progress in the use of integrin alpha(v)beta(3) antagonists and integrin-targeted delivery systems as potential cancer therapeutics, and discuss future perspectives.


Total retinal volumetric blood flow rate in diabetic patients with poor glycemic control.
Total retinal volumetric blood flow rate was measured in 12 normal subjects and 18 poorly controlled diabetic patients with background diabetic retinopathy. Maximum or center-line erythrocyte velocity (Vmax) was assessed by bidirectional laser Doppler velocimetry in four to five major retinal veins of one eye of each subject. Venous diameter (D) was measured from monochromatic fundus photographs. Total venous cross-section and measured total retinal volumetric blood flow in the diabetic patients were significantly larger than normal (P = 0.001 and P = 0.02, respectively). A positive linear correlation was found between Vmax and D in normal and diabetic eyes. Volumetric blood flow rate, Q, varied with D at a power of 2.87 in normal eyes. Total volumetric blood flow correlated with total venous cross-section. It was found that Q in the temporal retina was significantly larger than in the nasal retina in normal subjects (P = 0.0008) and diabetic patients (P = 0.0002). A significant difference in Q was observed between the superior and inferior retina in diabetic patients (P = 0.03) but not in normal subjects. The retinal vascular regulatory response to 100% oxygen breathing was reduced (P = 0.019) in diabetic patients and correlated with the level of background diabetic retinopathy. A close estimate of total volumetric blood flow may be obtained from blood flow measurement in one major retinal vein and the determination of total venous cross-section. This may be important for clinical studies in which measurements of all individual retinal veins may not be feasible.


Characterization of human MMTV-like (HML) elements similar to a sequence that was highly expressed in a human breast cancer: further definition of the HML-6 group.
Previously, we found a retroviral sequence, HML-6.2BC1, to be expressed at high levels in a multifocal ductal breast cancer from a 41-year-old woman who also developed ovarian carcinoma. The sequence of a human genomic clone (HML-6.28) selected by high-stringency hybridization with HML-6.2BC1 is reported here. It was 99% identical to HML-6.2BC1 and gave the same restriction fragments as total DNA. HML-6.28 is a 4.7-kb provirus with a 5'LTR, truncated in RT. Data from two similar genomic clones and sequences found in GenBank are also reported. Overlaps between them gave a rather complete picture of the HML-6.2BC1-like human endogenous retroviral elements. Work with somatic cell hybrids and FISH localized HML-6.28 to chromosome 6, band p21, close to the MHC region. The causal role of HML-6.28 in breast cancer remains unclear. Nevertheless, the ca. 20 Myr old HML-6 sequences enabled the definition of common and unique features of type A, B, and D (ABD) retroviruses. In Gag, HML-6 has no intervening sequences between matrix and capsid proteins, unlike extant exogenous ABD viruses, possibly an ancestral feature. Alignment of the dUTPase showed it to be present in all ABD viruses, but gave a phylogenetic tree different from trees made from other ABD genes, indicating a distinct phylogeny of dUTPase. A conserved 24-mer sequence in the amino terminus of some ABD envelope genes suggested a conserved function.


ACOG Committee Opinion No. 758 Summary: Promoting Healthy Relationships in Adolescents.
Obstetrician-gynecologists have the opportunity to promote healthy relationships by encouraging adolescents to discuss past and present relationships while educating them about respect for themselves and mutual respect for others. Because middle school is a time when some adolescents may develop their first romantic or sexual relationships, it is an ideal timeframe for obstetrician-gynecologists and other health care providers, parents, and guardians to play a role in anticipatory guidance. Creating a nonjudgmental environment and educating staff on the unique concerns of adolescents are helpful ways to provide effective and appropriate care to this group of patients. Obstetrician-gynecologists and other health care providers caring for minors should be aware of federal and state laws that affect confidentiality. Obstetrician-gynecologists should screen patients routinely for intimate partner violence along with reproductive and sexual coercion and be prepared to address positive responses. Furthermore, obstetrician-gynecologists should be aware of mandatory reporting laws in their state when intimate partner violence, adolescent dating violence, or statutory rape is suspected. Pregnant and parenting adolescents; lesbian, gay, bisexual, transgender, queer or questioning (LGBTQ) individuals; and adolescents with physical and mental disabilities are at particular risk of disparities in the health care system. The promotion of healthy relationships in these groups requires the obstetrician-gynecologist to be aware of the unique barriers and hurdles to sexual and nonsexual expression, as well as to health care. Interventions to promote healthy relationships and a strong sexual health framework are more effective when started early and can affect indicators of long-term individual health and public health.


Cutaneous and subcutaneous metastases from bladder carcinoma.
Cutaneous metastases of bladder carcinoma are very rare. The rate of cutaneous metastasis in urologic malignancies is 1.3% and this rate was found to be 0.84% for bladder carcinomas. Cutaneous metastasis of bladder carcinoma can be confused with cellulitis. This case report presents a 60-year-old patient operated on for bladder carcinoma and undergoing treatment for cellulitis because of erythematous lesions appearing in the suprapubic region in the early postoperative period. As there was no response to antibiotic treatment, skin biopsy was performed and cutaneous metastasis was diagnosed. Subsequently, chemotherapy with carboplatin and gemcitabine was started. The prognosis of bladder carcinoma with cutaneous metastasis has been reported to be poor.


Phonomicrosurgical management of vocal fold polyps: the subepithelial microflap resection technique.
Vocal fold polyps are typically caused by acute and chronic trauma to the microvasculature of the superficial lamina propria (SLP). Shearing stresses that are induced by hyperfunctional glottal sound production lead to bleeding into the SLP and malformed neo-vascularized masses. Because the primary process does not involve the epithelium, the authors designed a technique to resect hemorrhagic polyps by epithelial cordotomy with partial or complete preservation of the vocal fold epithelium. This approach is different from the traditional microsurgical resection of hemorrhagic polyps by amputation with or without the carbon dioxide laser. Forty patients who underwent microlaryngoscopic resection of hemorrhagic polyps from 1996 through 1998 were reviewed retrospectively. Thirty-six of the 40 procedures were by epithelial cordotomy and subepithelial removal of the polyp contents. Sixteen of 36 were assisted by a subepithelial infusion of saline and epinephrine, and all were 3 mm to 6 mm. Four of 40 polyps were amputated; all of these were less than 3 mm and were pedicled on a narrow base. Cold instruments were used exclusively in all 40 patients. Postoperative laryngeal stroboscopy within 2 weeks revealed improved mucosal wave propagation and improved glottal closure in all 33 patients in whom postoperative strobovideolaryngoscopy was available. The epithelial cordotomy technique was introduced to minimize disturbance of normal SLP and epithelium. Despite the hemorrhagic nature of these lesions, cold instruments could be used exclusively with facility due to careful microdissection between the polyp and the residual normal SLP and the enhanced hemostasis provided by the subepithelial infusion of saline and epinephrine. The rapid return to improved glottal function is the result of this ultra tissue-sparing technique.


Diagnostic Performance of Plasmodium falciparum Histidine-Rich Protein-2 Antigen-Specific Rapid Diagnostic Test in Children at the Peripheral Health Care Level in Nanoro (Burkina Faso).
(1) Background: Malaria control has strongly benefited from the implementation of rapid diagnostic tests (RDTs). The malaria RDTs used in Burkina Faso, as per the recommendation of the National Malaria Control Program, are based on the detection of histidine-rich protein-2 (PfHRP2) specific to Plasmodium falciparum, which is the principal plasmodial species causing malaria in Burkina Faso. However, there is increasing concern about the diagnostic performance of these RDTs in field situations, and so constant monitoring of their accuracy is warranted. (2) Methods: A prospective study was performed in the health district of Nanoro, where 391 febrile children under 5 years with an axillary temperature ≥37.5 °C presenting at participating health facilities were subjected to testing for malaria. The HRP2-based RDT and expert microscopy were used to determine the diagnostic performance of the former. Retrospectively, the correctness of the antimalaria prescriptions was reviewed. (3) Results: Taking expert malaria microscopy as the gold standard, the sensitivity of the employed RDT was 98.5% and the specificity 40.5%, with a moderate agreement between the RDT testing and microscopy. In total, 21.7% of cases received an inappropriate antimalarial treatment based on a retrospective assessment with expert microscopy results. (4) Conclusion: Malaria remains one of the principal causes of febrile illness in Burkina Faso. Testing with HRP2-based RDTs is inaccurate, in particular, due to the low specificity, which results in an over-prescription of antimalarials, with emerging antimalarial drug resistance as an important risk and many children not being treated for potential other causes of fever.


Effect of porcine fetal enamel matrix derivative on chondrocyte proliferation, differentiation, and local factor production is dependent on cell maturation state.
Recent studies have shown that porcine fetal enamel matrix derivative (EMD) can enhance the osteoinductive ability of demineralized freeze-dried bone allograft (DFDBA) in a nude mouse muscle implantation model. This suggests that one or more components of EMD can regulate the process of endochondral ossification initiated by DFDBA. To substantiate this hypothesis, in the current study, chondrocytes in the endochondral pathway at two stages of maturation were tested for their response to EMD. Chondrocytes were isolated from the resting zone and growth zone (prehypertrophic and upper hypertrophic cell zones) of the costochondral growth plate cartilage of adolescent rats. The results showed that the relatively immature resting zone cells responded to EMD with an increase in proliferation and a decrease in differentiation as measured by alkaline-phosphatase-specific activity. In addition, EMD stimulated a fivefold increase in PGE(2) production, but was without effect on collagen synthesis, proteoglycan sulfation, and TGF-beta(1) production. The more mature growth zone cells also responded to EMD with increased proliferation, but alkaline-phosphatase-specific activity was unchanged, and there was only a modest increase in PGE(2) production. In contrast to resting zone cells, growth zone cells exhibited a decrease in collagen synthesis, proteoglycan sulfation, and TGF-beta(1) production. These observations indicate that EMD has prominent effects on cells in the endochondral pathway. In particular, EMD stimulates the production of more cells, but inhibits their maturation. This would increase the pool of cells available for subsequent differentiation in response to other factors.


Successful treatment of delusions of parasitosis with olanzapine.
Delusional parasitosis is a rare disorder in which patients have a fixed, false belief of being infested with parasites. It is often accompanied by a refusal to seek psychiatric care. Delusions of parasitosis is classically treated with typical antipsychotic agents, the traditional dermatologic choice being pimozide. However, pimozide's adverse effect profile and the need for frequent electrocardiographic monitoring make such treatment less practical. We describe 3 patients who were diagnosed as having delusional parasitosis that was successfully treated with a recently Food and Drug Administration-approved atypical antipsychotic agent, olanzapine (5 mg/d). Olanzapine has a more benign adverse effect profile than typical antipsychotic agents and eliminates the need for electrocardiographic monitoring. Olanzapine therapy has been associated with such adverse effects as sedation, hyperlipidemia, weight gain, and insulin resistance, all of which were infrequent in our patients. Olanzapine is an atypical antipsychotic agent that can be used as a first-line agent in delusional parasitosis as a safer therapeutic option without a specialized monitoring regimen.


Quantitative assessment of the sulfuric acid contribution to new particle growth.
The Nano Aerosol Mass Spectrometer (NAMS) was deployed to rural/coastal and urban sites to measure the composition of 20-25 nm diameter nanoparticles during new particle formation (NPF). NAMS provides a quantitative measure of the elemental composition of individual, size-selected nanoparticles. In both environments, particles analyzed during NPF were found to be enhanced in elements associated with inorganic species (nitrogen, sulfur) relative to that associated with organic species (carbon). A molecular apportionment algorithm was applied to the elemental data in order to place the elemental composition into a molecular context. These measurements show that sulfate constitutes a substantial fraction of total particle mass in both environments. The contribution of sulfuric acid to new particle growth was quantitatively determined and the gas-phase sulfuric acid concentration required to incorporate the measured sulfate fraction was calculated. The calculated values were compared to those calculated by a sulfuric acid proxy that considers solar radiation and SO(2) levels. The two values agree within experimental uncertainty. Sulfate accounts for 29-46% of the total mass growth of particles. Other species contributing to growth include ammonium, nitrate, and organics. For each location, the relative amounts of these species do not change significantly with growth rate. However, for the coastal location, sulfate contribution increases with increasing temperature whereas nitrate contribution decreases with increasing temperature.


Safety and Efficacy of Oral and/or Intravenous Tedizolid Phosphate From a Randomized Phase 3 Trial in Adolescents With Acute Bacterial Skin and Skin Structure Infections.
Tedizolid phosphate is an oxazolidinone prodrug approved in 2014 for treatment of adults with acute bacterial skin and skin structure infections (ABSSSIs); however, efficacy has not previously been evaluated in children. This study compared the safety and efficacy of tedizolid (administered as tedizolid phosphate) with active antibacterial comparators for the treatment of ABSSSIs in adolescents. This was a randomized, assessor-blind, global phase 3 study of tedizolid versus active comparators for the treatment of Gram-positive ABSSSIs in adolescents (12 to <18 years of age; NCT02276482). Enrolled participants were stratified by region and randomized 3:1 to receive tedizolid phosphate 200 mg (oral and/or intravenous) once daily for 6 days or active comparator, selected by investigator from an allowed list per local standard of care, for 10 days. The primary endpoint was safety; blinded investigator's assessment of clinical success at the test-of-cure visit (18-25 days after the first dose) was a secondary efficacy endpoint. Statistical comparisons between treatment groups were not performed. Of the 121 participants enrolled, 120 were treated (tedizolid, n = 91; comparator, n = 29). Treatment-emergent adverse events were balanced between treatment groups (tedizolid, 14.3%; comparator, 10.3%). Overall, 3 participants (3.3%) in the tedizolid group and 1 (3.4%) in the comparator group experienced a single drug-related TEAE. Clinical success rates were high in both treatment groups: 96.7% and 93.1% at the test-of-cure visit for the tedizolid and comparator groups, respectively. Tedizolid demonstrated safety and efficacy similar to comparators for the treatment of ABSSSIs in adolescents.


Clinical framework for next generation sequencing based analysis of treatment predictive mutations and multiplexed gene fusion detection in non-small cell lung cancer.
Precision medicine requires accurate multi-gene clinical diagnostics. We describe the implementation of an Illumina TruSight Tumor (TST) clinical NGS diagnostic framework and parallel validation of a NanoString RNA-based ALK, RET, and ROS1 gene fusion assay for combined analysis of treatment predictive alterations in non-small cell lung cancer (NSCLC) in a regional healthcare region of Sweden (Scandinavia). The TST panel was clinically validated in 81 tumors (99% hotspot mutation concordance), after which 533 consecutive NSCLCs were collected during one-year of routine clinical analysis in the healthcare region (~90% advanced stage patients). The NanoString assay was evaluated in 169 of 533 cases. In the 533-sample cohort 79% had 1-2 variants, 12% >2 variants and 9% no detected variants. Ten gene fusions (five ALK, three RET, two ROS1) were detected in 135 successfully analyzed cases (80% analysis success rate). No ALK or ROS1 FISH fusion positive case was missed by the NanoString assay. Stratification of the 533-sample cohort based on actionable alterations in 11 oncogenes revealed that 66% of adenocarcinomas, 13% of squamous carcinoma (SqCC) and 56% of NSCLC not otherwise specified harbored ≥1 alteration. In adenocarcinoma, 10.6% of patients (50.3% if including KRAS) could potentially be eligible for emerging therapeutics, in addition to the 15.3% of patients eligible for standard EGFR or ALK inhibitors. For squamous carcinoma corresponding proportions were 4.4% (11.1% with KRAS) vs 2.2%. In conclusion, multiplexed NGS and gene fusion analyses are feasible in NSCLC for clinical diagnostics, identifying notable proportions of patients potentially eligible for emerging molecular therapeutics.


Initial coupling of binding to gating mediated by conserved residues in the muscle nicotinic receptor.
We examined functional consequences of intrasubunit contacts in the nicotinic receptor alpha subunit using single channel kinetic analysis, site-directed mutagenesis, and structural modeling. At the periphery of the ACh binding site, our structural model shows that side chains of the conserved residues alphaK145, alphaD200, and alphaY190 converge to form putative electrostatic interactions. Structurally conservative mutations of each residue profoundly impair gating of the receptor channel, primarily by slowing the rate of channel opening. The combined mutations alphaD200N and alphaK145Q impair channel gating to the same extent as either single mutation, while alphaK145E counteracts the impaired gating due to alphaD200K, further suggesting electrostatic interaction between these residues. Interpreted in light of the crystal structure of acetylcholine binding protein (AChBP) with bound carbamylcholine (CCh), the results suggest in the absence of ACh, alphaK145 and alphaD200 form a salt bridge associated with the closed state of the channel. When ACh binds, alphaY190 moves toward the center of the binding cleft to stabilize the agonist, and its aromatic hydroxyl group approaches alphaK145, which in turn loosens its contact with alphaD200. The positional changes of alphaK145 and alphaD200 are proposed to initiate the cascade of perturbations that opens the receptor channel: the first perturbation is of beta-strand 7, which harbors alphaK145 and is part of the signature Cys-loop, and the second is of beta-strand 10, which harbors alphaD200 and connects to the M1 domain. Thus, interplay between these three conserved residues relays the initial conformational change from the ACh binding site toward the ion channel.


Nutritional intervention with TGF-beta enriched food for special medical purposes (TGF-FSMP) is associated with a reduction of malnutrition, acute GVHD, pneumonia and may improve overall survival in patients undergoing allogeneic hematopoietic stem transplantation.
Malnutrition in allogeneic stem cell transplant (allo-SCT) is associated with poor outcomes. Supplementation with Foods for Special Medical Purposes may be a valid alternative to enteral nutrition or total parental nutrition to reduce malnutrition in allo-SCT. In this study, 133 patients consecutively allo-transplanted were assessed for nutritional status by Patient- Generated Subjective Global Assessment (PG-SGA) and supplemented with TGF-beta enriched Food for Special Medical Purposes (TGF-FSMP). PG-SGA, gold standard for nutritional assessment in oncologic patients, was assessed at admission and on day 0, +7, +14, +21, and + 28 from transplant and categorized as follows: A = good nutritional status; B = moderate malnutrition; C = severe malnutrition. TGF-FSMP (Modulen-IBD) is currently used in Inflammatory Bowel Diseases (IBD) as primary nutritional support and in this study the dose was calculated according to BMI and total daily energy expenditure (TDEE). The patients assuming ≥50% of the prescribed TGF-FSMP dose were classified in Group A; the patients who received < 50% were included in Group B per protocol. The primary endpoint of the study was the assessment of the malnourished patients in Group A and B at day+28 after transplantation, according to the criteria of PG-SGA C categorization. At day +28 after transplant: i) patients in Group A were significantly less severely malnourished than patients in the Group B (21/76,28% vs 42/53, 79% respectively, OR 2.86 - CI 1.94-4.23 -, p = 0.000); ii) the incidence of severe (MAGIC II-IV) aGVHD and of any grade gastrointestinal (GI) aGVHD was higher in Group B than in Group A, (43% vs 21% p = 0.003) and (34.5% vs 9.2% p = 0.001); iii) Pneumonia was more frequent in the malnourished patients of Group B than in well/moderate nourished patients of Group A (52.7% vs 27.6% p = 0.002). In group A parenteral nutrition was avoided more frequently than in group B (67.5% vs 33.3% p = 0.000) and a median hospital stay of 27 days in comparison to 32 was reported (p = 0.006). The estimated median overall survival (OS) of the population was 33 months in Group A and 25.1 months in group B (p = 0.03). By multivariate and ANN analysis, TGF-FSMP TR < 50% assumption was significantly correlated with malnutrition, severe and GI aGVHD, pneumonia and reduced OS.


[Apoptosis-like cell death of Cryptococcus neoformans mediated by Staphylococcus aureus contact].
Co-culture of the fungal pathogen Cryptococcus neoformans with Staphylococcus aureus results in the death of the fungus, caused by the adherence to the latter. The present study found that the molecules responsible for this adherence were capsular glucuronoxylomannan (GXM) (present on C. neoformans) and a glycolytic enzyme triosephosphate isomerase (TPI) (present on S. aureus). The mannan backbone of GXM and purified TPI interacted in vitro. GXM-bound TPI molecules were identified by immunoelectron microscopy. The death of C. neoformans was accompanied by decreased actin turnover, increased accumulation of reactive oxygen species, and DNA fragmentation. This process may also be influenced by the Rho/Rho-associated coiled-coil-forming kinase (ROCK) pathway and enhanced expression of voltage-dependent ion-selective channels. Taken together, these results suggest that Rho-ROCK signaling may play a role via the mitochondrial pathway in the induction of C. neoformans apoptosis-like cell death after its adherence to S. aureus adherence.


beta 2-Glycoprotein I and anti-beta 2-glycoprotein I antibodies: where are we now?
beta 2-Glycoprotein I (beta 2-GPI), a plasma protein with in vitro anticoagulant properties, has been recognized to have an important role in the antiphospholipid syndrome (APS) as a cofactor and an (co)antigen in ELISA assays. Although beta 2-GPI levels were found to be increased in some patients with APS, the clinical value of measuring beta 2-GPI levels in APS is not known. Several reports have suggested that anti-beta 2-GPI antibodies may be a marker for the APS and might be more specific for the vascular complications of the APS than anticardiolipin antibodies. There have been major discoveries about phospholipid (PL) and antibody binding sites on beta 2-GPI, although more studies are needed. Reports of changes in cell membrane PL composition or exposure of other anionic molecules by apoptosis, cell activation and oxidative injury suggest mechanisms to explain beta 2-GPI binding and the generation of cryptic epitopes for aPL/anti-beta 2-GPI antibodies.


Tumor-associated CD75s- and iso-CD75s-gangliosides are potential targets for adjuvant therapy in pancreatic cancer.
Pancreatic adenocarcinoma confers one of the highest mortality rates in malignant human tumors with very poor prognosis. Because as yet no treatments are available that produce a substantial survival benefit for this fatal neoplasia, new therapeutic concepts are urgently required to support cancer standard treatment. In search of tumor-associated gangliosides with therapeutic background, we probed a random collection of cancerous and adjacent normal postoperative tissue samples from 38 patients for the expression of CD75s- and iso-CD75s-gangliosides. We exhaustively analyzed the expression of CD75s-1-ganglioside (IV(6)Neu5Ac-nLc4Cer) and structurally closely related iso-CD75s-1-ganglioside (IV(3)Neu5Ac-nLc4Cer) by means of immunohistology of cryosections and semiquantitative TLC of tissue lipid extracts combined with mass spectrometry. CD75s-1- and iso-CD75s-1-ganglioside showed an elevated expression in 42% and 66% of the tumors, respectively, indicating a significant association with neoplastic transformation (P = 0.001). Thus, increased expression of CD75s-1- and iso-CD75s-1-gangliosides renders these cell surface molecules promising candidates for oncologic applications. Further statistical analysis revealed a significant enhancement of CD75s-1-ganglioside in the group of less differentiated tumors (grade >2) suggesting this ganglioside as a potential marker for poor differentiation. The CD75s-specific antitumor drug rViscumin, which represents the recombinant counterpart of the ribosome-inactivating lectin viscumin, has successfully passed clinical phase I trials and provides an opportunity for treating pancreatic cancer. Consequently, if an enhanced expression is existent in malignant tissues, we propose the targeting of CD75s-gangliosides with rViscumin as a novel potential strategy in adjuvant treatment of pancreatic malignancies.


Blood and spleen lymphocytes as targets for immunotoxic effects in the rat--a comparison.
Traditionally, immunotoxicological studies in the rat have been performed by measuring the effect of chemical substances on spleen lymphocytes in vivo and in vitro. However, rat blood lymphocytes may be more relevant than spleen cells for comparison with human blood lymphocytes. Further, lymphocytes in blood may be a more sensitive indicator of immunotoxic effects than spleen lymphocytes. Finally, in longitudinal studies peripheral blood specimens can be collected repeatedly from the same animals, thereby reducing the number of animals sacrificed and, possibly, experimental variation. We compared blood and spleen lymphocyte parameters in rats treated with a single dose of the immunosuppressant cyclophosphamide (CY), monitoring effects on blood and spleen lymphocytes by immunophenotyping. We also performed repeated bleedings to demonstrate the feasibility of following the time course of induced changes in the same animals. Immunophenotyping as well as total mononuclear cell counts consistently showed as large or lager effects of CY in blood lymphocytes than in spleen cells. Further, the measured effects in blood lymphocytes became statistically significant at an earlier time point, compared to spleen cells. Repeated bleedings of the same animals illustrated that blood specimens drawn from a peripheral vein give sufficient numbers of cells to perform immunotoxicological tests.


Primary hyperparathyroidism in adults-(Part I) assessment and medical management: Position statement of the endocrine society of Australia, the Australian & New Zealand endocrine surgeons, and the Australian & New Zealand bone and mineral society.
To formulate clinical consensus recommendations on the presentation, assessment, and management of primary hyperparathyroidism (PHPT) in adults. Representatives from relevant Australian and New Zealand Societies used a systematic approach for adaptation of guidelines (ADAPTE) to derive an evidence-informed position statement addressing nine key questions. PHPT is a biochemical diagnosis. Serum calcium should be measured in patients with suggestive symptoms, reduced bone mineral density or minimal trauma fractures, and in those with renal stones. Other indications are detailed in the manuscript. In patients with hypercalcaemia, intact parathyroid hormone, 25-hydroxy vitamin D, phosphate, and renal function should be measured. In established PHPT, assessment of bone mineral density, vertebral fractures, urinary tract calculi/nephrocalcinosis and quantification of urinary calcium excretion is warranted. Parathyroidectomy is the only definitive treatment and is warranted for all symptomatic patients and should be considered for asymptomatic patients without contraindications to surgery and with >10 years life expectancy. In patients who do not undergo surgery, we recommend annual evaluation for disease progression. Where the diagnosis is not clear or the risk-benefit ratio is not obvious, multidisciplinary discussion and formulation of a consensus management plan is appropriate. Genetic testing for familial hyperparathyroidism is recommended in selected patients. These clinical consensus recommendations were developed to provide clinicians with contemporary guidance on the assessment and management of PHPT in adults. It is anticipated that improved health outcomes for individuals and the population will be achieved at a decreased cost to the community.


Can brief behavioral health interventions reduce suicidal and self-harm ideation in primary care patients?
We examined whether brief behavioral health visits reduced suicidal and self-harm ideation among primary care patients and compared the effectiveness of interventions that targeted ideation directly (i.e., safety planning) with those that targeted ideation indirectly through management of underlying mental illness (e.g., behavioral activation). We examined first- and last-visit data from 31 primary care patients with suicidal or self-harm ideation seen by behavioral health consultants. Patients reported significantly lower frequencies of suicidal and self-harm ideation at their final visit than at their initial visit. Patients whose ideation was targeted directly showed greater improvements than patients whose ideation was targeted indirectly. Although preliminary, results suggest mild to moderate suicidal ideation could be addressed in primary care through integration of behavioral health consultants into the medical team. (PsycINFO Database Record


Personal recognition using hand shape and texture.
This paper proposes a new bimodal biometric system using feature-level fusion of hand shape and palm texture. The proposed combination is of significance since both the palmprint and hand-shape images are proposed to be extracted from the single hand image acquired from a digital camera. Several new hand-shape features that can be used to represent the hand shape and improve the performance are investigated. The new approach for palmprint recognition using discrete cosine transform coefficients, which can be directly obtained from the camera hardware, is demonstrated. None of the prior work on hand-shape or palmprint recognition has given any attention on the critical issue of feature selection. Our experimental results demonstrate that while majority of palmprint or hand-shape features are useful in predicting the subjects identity, only a small subset of these features are necessary in practice for building an accurate model for identification. The comparison and combination of proposed features is evaluated on the diverse classification schemes; naive Bayes (normal, estimated, multinomial), decision trees (C4.5, LMT), k-NN, SVM, and FFN. Although more work remains to be done, our results to date indicate that the combination of selected hand-shape and palmprint features constitutes a promising addition to the biometrics-based personal recognition systems.


Capsule endoscopy: past, present, and future.
A capsule endoscope is a swallowable wireless miniature camera for getting images of the gastrointestinal (GI) mucosa. The initial capsule endoscope model was developed by Given Imaging and approved in Western countries in 2001. Before the introduction of capsule endoscopy (CE) and double-balloon endoscopy (DBE), there was no effective modality for the evaluation and management of patients with obscure GI bleeding. Obscure GI bleeding is defined as bleeding of unknown origin that persists or recurs after a negative initial or primary endoscopy (colonoscopy or upper endoscopy) result. The first capsule endoscope model, which is now regarded as a first-line tool for the detection of abnormalities of the small bowel, was the PillCam SB. It was approved in Japan in April 2007. The main indication for use of the PillCam SB is obscure GI bleeding. Almost the only complication of CE is capsule retention, which is the capsule remaining in the digestive tract for a minimum of 2 weeks. A retained capsule can be retrieved by DBE. There are some limitations of CE in that it cannot be used to obtain a biopsy specimen or for endoscopic treatment. However, the combination of a PillCam SB and DBE seems to be the best strategy for management of obscure GI bleeding. Recently, several new types of capsule endoscope have been developed, such as Olympus CE for the small bowel, PillCam ESO for investigation of esophageal diseases, and PillCam COLON for detection of colonic neoplasias. In the near future, CE is expected to have a positive impact on many aspects of GI disease evaluation and management.


Risk determination for pancreatic cancer.
Pancreatic cancer represents one of the leading causes of cancer related deaths worldwide and constitutes a major public health problem. Despite the advances in diagnosis and treatment, the overall five-year survival remains low, thus leading the focus of medical research towards the identification and modification of potential risk factors. This year, in ASCO Annual Meeting two interesting studies were presented. Ghani et al. (abstract #e15183) sought to investigate the effect of smoking on chemotherapy response in patients with metastatic pancreatic cancer, while Walker et al. (abstract #4117) presented the results of their study regarding the effect of statin use in the prevention of pancreatic cancer. Both studies concluded to useful results that along with the existing literature may further stimulate medical research towards better recognition of risk factors and the application of this knowledge in the clinical practice.


The somatotopic organization of the cat trigeminal ganglion as determined by the horseradish peroxidase technique.
The somatotopic organization of the cat trigeminal ganglion has been investigated in the present study by using the horseradish peroxidase (HRP) technique. In separate animals, the corneal, supraorbital, infraorbital, inferior alveolar, or mental branches of the trigeminal nerve have been transected and then soaked in concentrated solutions of HRP. Retrogradely labeled corneal and supraorbital neurons have been found, with extensive overlap between the two cell populations, in the anteromedial region of the trigeminal ganglion. Inferior alveolar and mental neurons have been found to possess similar distributions within the posterolateral part of the trigeminal ganglion. Infraorbital cells have been localized in a central position. The cell bodies of any given nerve are found in at least minimal numbers in all dorsoventral levels of the trigeminal ganglion. However, cell bodies of origin of the supraorbital nerve and the lateral branch of the infraorbital nerve, innervating more posterior or lateral areas of the head and face, are found in greater numbers dorsally. Conversely, cell bodies of origin of the medial branch of the infraorbital nerve, the inferior alveolar nerve, and the mental nerve, supplying more rostral or intraoral areas of the orofacial region, are present in greater numbers ventrally. In contrast, corneal neurons are distributed uniformly in the dorsoventral axis. The ophthalmic and maxillary regions of the trigeminal ganglion appear to be well segregated, whereas the maxillary and mandibular regions exhibit a somewhat greater degree of overlap. Cell bodies of corneal afferent neurons range from 20 to 50 micrometer in diameter, whereas those of supraorbital, infraorbital, inferior alveolar and mental neurons measure from 20 to 85 micrometer. It is concluded from the findings of the present work that much of the cat trigeminal ganglion is organized somatotopically in not only the mediolateral axis but also in the dorsoventral axis.


Haploinsufficiency of Parp1 accelerates Brca1-associated centrosome amplification, telomere shortening, genetic instability, apoptosis, and embryonic lethality.
The breast tumor associated gene-1 (BRCA1) and poly(ADP-ribose) polymerase-1 (PARP1) are both involved in DNA-damage response and DNA-damage repair. Recent investigations have suggested that inhibition of PARP1 represents a promising chemopreventive/therapeutic approach for specifically treating BRCA1- and BRCA2-associated breast cancer. However, studies in mouse models reveal that Parp1-null mutation results in genetic instability and mammary tumor formation, casting significant doubt on the safety of PARP1 inhibition as a therapy for the breast cancer. To study the genetic interactions between Brca1 and Parp1, we interbred mice carrying a heterozygous deletion of full-length Brca1 (Brca1(+/Delta11)) with Parp1-null mice. We show that Brca1(Delta11/Delta11);Parp1(-/-) embryos die before embryonic (E) day 6.5, whereas Brca1(Delta11/Delta11) embryos die after E12.5, indicating that absence of Parp1 dramatically accelerates lethality caused by Brca1 deficiency. Surprisingly, haploinsufficiency of Parp1 in Brca1(Delta11/Delta11) embryos induces a severe chromosome aberrations, centrosome amplification, and telomere dysfunction, leading to apoptosis and accelerated embryonic lethality. Notably, telomere shortening in Brca1(Delta11/Delta11);Parp1(+/-) MEFs was correlated with decreased expression of Ku70, which plays an important role in telomere maintenance. Thus, haploid loss of Parp1 is sufficient to induce lethality of Brca1-deficient cells, suggesting that partial inhibition of PARP1 may represent a practical chemopreventive/therapeutic approach for BRCA1-associated breast cancer.


Are undocumented migrants' entitlements and barriers to healthcare a public health challenge for the European Union?
Undocumented migrants (UMs) are at higher risk for health problems because of their irregular status and the consequences of economic and social marginalization. Moreover, the emergent reality of undocumented migration in Europe calls for action in the field of management of UM's health demands as their access to health services has become a sensitive political and social issue. In this light, this paper aims to address UMs' entitlement and barriers to healthcare and related policies citing evidence from peer-reviewed and grey literature concerning people living in a country within the European Union without the legal right to be/remain in the destination country. A systematic review was performed using several databases and websites, and a total of 54 publications in English, with full text available, were taken into consideration. Between 2000 and 2015, Europe hosted the second largest number of international migrants (20 million, 1.3 million per year) after Asia. Even though there is limited evidence specifically focused on UMs' health, it is possible to state that infectious diseases, chronic illnesses, mental disorders, maternal-child conditions, dental issues, acute illnesses and injuries are the most common pathologies. In most cases across Europe, UMs have access only to emergency care. Even in countries where they are fully entitled to healthcare, formal and informal barriers hinder them from being or feeling entitled to this right. Socio-cultural barriers, such as language and communication problems, lack of formal and informal social and healthcare networks and lack of knowledge about the healthcare system and about informal networks of healthcare professionals are all common impediments. From the healthcare providers' perspective, there can be difficulties in providing appropriate care and in dealing with cultural and language barriers and false identification. Communication strategies play a central role in addressing the inequalities in access to healthcare services, and the definition and provision of specific training, focused on UMs' health needs, would be desirable. Improving access to healthcare for UMs is an urgent priority since the lack of access is proven to have serious consequences for UMs' health and wellbeing. Notwithstanding, few available examples of policies and best practices aimed at overcoming barriers in the delivery of healthcare to UMs are available.


Interaction between gonadotropin-releasing hormone and bone morphogenetic protein-6 and -7 signaling in LβT2 gonadotrope cells.
It is known that bone morphogenetic proteins (BMPs) regulate gonadotropin transcription and production by pituitary gonadotrope cells. However, the role of BMPs in gonadotropin-releasing hormone (GnRH)-induced FSH production remains uncertain. Here, we describe a functional link between BMP-6 and BMP-7 signals and FSH transcriptional activity induced by GnRH using mouse gonadotrope LβT2 cells. In LβT2 cells, BMP-6 and BMP-7 increased mouse FSHβ-promoter activity in a concentration-dependent manner. The induction by BMP-6 and BMP-7 was inhibited by treatment with extracellular domains of ActRII but not BMPRII. These findings suggest that the type II receptor ActRII participates in BMP-induced FSHβ transcription regulation. Notably, BMP-6, but not BMP-7, enhanced GnRH-induced FSHβ-promoter activity in LβT2 cells. Since GnRH stimulated MAPK phosphorylation in LβT2 cells, a functional link between MAPK and FSHβ transcription was examined. Inhibition of the ERK pathway, but not that of p38 or SAPK/JNK signaling, suppressed GnRH-induced FSHβ transcription, suggesting that ERK is functionally involved in GnRH-induced FSHβ transcription. Co-treatment with BMP-7, but not with BMP-6, suppressed GnRH-induced MAPK phosphorylation in LβT2 cells. Thus, the difference between BMP-6 and BMP-7 in enhancing GnRH-induced FSHβ transcription may be due to the differential effects of BMP ligands on GnRH-induced ERK signaling. On the other hand, GnRH reduced Smad1/5/8 phosphorylation but increased Smad6/7 expression. These findings imply the presence of a functional link between GnRH action, MAPK signaling and the BMP system in pituitary gonadotropes for fine-tuning of FSH gene expression.


Impact of Pandemics/Epidemics on Emergency Department Utilization for Mental Health and Substance Use: A Rapid Review.
Background: A prolonged COVID-19 pandemic has the potential to trigger a global mental health crisis increasing demand for mental health emergency services. We undertook a rapid review of the impact of pandemics and epidemics on emergency department utilization for mental health (MH) and substance use (SU). Objective: To rapidly synthesize available data on emergency department utilization for psychiatric concerns during COVID-19. Methods: An information specialist searched Medline, Embase, Psycinfo, CINAHL, and Scopus on June 16, 2020 and updated the search on July 24, 2020. Our search identified 803 abstracts, 7 of which were included in the review. Six articles reported on the COVID-19 pandemic and one on the SARS epidemic. Results: All studies reported a decrease in overall and MH related ED utilization during the early pandemic/epidemic. Two studies found an increase in SU related visits during the same period. No data were available for mid and late stage pandemics and the definitions for MH and SU related visits were inconsistent across studies. Conclusions: Our results suggest that COVID-19 has resulted in an initial decrease in ED visits for MH and an increase in visits for SU. Given the relative paucity of data on the subject and inconsistent analytic methods used in existing studies, there is an urgent need for investigation of pandemic-related changes in ED case-mix to inform system-level change as the pandemic continues.


Effect of pH on phosphate transport into intestinal brush-border membrane vesicles.
The effect of pH on the rate of phosphate (Pi) uptake was studied in rabbit duodenal brush-border membrane vesicles. Pi uptake was found to be sodium dependent at all pH values tested (5.7-8.1). Further, the rate of Pi uptake depended on pH; for instance, with 100 mM external sodium, reducing the pH from 8.1 to 6.8 or 5.7 doubled the rate of Pi influx. At 100 mM external sodium, experiments under initial rate conditions, carried out with varying Pi concentrations and at pH values of 6, 6.8, or 7.6, showed that sodium-dependent Pi uptake was saturable at the three pH values tested; the apparent Km expressed in function of total Pi was not dependent on pH. Vmax was not affected between pH 6 and 6.8 but was significantly reduced at pH 7.6. Lowering external sodium lowered Vmax at all pH values investigated. At acid and alkaline pH the rate of Pi uptake was a sigmoidal function of the external sodium concentration. Hill coefficients, calculated from these experiments, exceeded unity and were unaffected by pH. At saturating sodium concentrations, the rate of Pi uptake was higher at pH 6 than at pH 7.6. The [Na]0.5 was lower at pH 7.6 than at pH 6. Further, sodium-dependent Pi uptake appeared to be electrogenic at acid and alkaline pH. It is concluded that the pH dependence of intestinal Pi transport is not an expression of preferential transport of monovalent or divalent phosphate. The pH dependence appears to reflect properties of the sodium-phosphate cotransport mechanism and is in part related to changes in the affinity of the transport system for sodium.


Effects of Gelatin Methacrylate Hydrogel on Corneal Repair and Regeneration in Rats.
This study investigates the repairing process of rat cornea after surgery of lamellar keratoplasty (LKP) and evaluates the effects of gelatin methacrylate (GelMA) hydrogel. In the LKP group, the lamellar stroma matrixes of Sprague-Dawley rats were transplanted to enhanced green fluorescent protein rats, whereas those in the GelMA group were also embedded with a GelMA hydrogel during the corneal transplantation. Grafted eyes were harvested on days seven, 30, and 90. Hematoxylin and eosin staining, immunofluorescence staining, scanning electron microscopy, optical coherence tomography, and a slit-lamp microscope were used to study the process of corneal restoration and regeneration. A total of 42 rats were analyzed, including 18 rats in each of the experimental group and six rats in the control group. After three months, the infiltration degree of inflammatory cells differed between the LKP group and the GelMA group (P < 0.001). Moreover, in multiple comparisons in corneal thickness, significant difference was observed between the LKP group and the GelMA group. There was also divergence in the results between the LKP group and the control group (P < 0.001, P < 0.001). At the same time, the expression of α-smooth muscle actin (α-SMA) and transforming growth factor (TGF)-β1 varied distinctly between the LKP group and the GelMA group (P < 0.05, P < 0.001). Significant differences were demonstrated between the LKP group and the GelMA group in inflammatory cell infiltration, corneal thickness, as well as the expression of α-SMA and TGF-β1. Those differences indicate the ability of GelMA hydrogel to support alleviation in corneal stroma fibrosis and show the influences of fibrosis in the dysfunction of corneal refractive power. Our research provides new ideas for the future development of LKP and tissue-engineered corneas.


Characterization of tetratricopeptide repeat-containing proteins critical for cilia formation and function.
Cilia formation and function require a special set of trafficking machinery termed intraflagellar transport (IFT), consisting mainly of protein complexes IFT-A, IFT-B, BBSome, and microtubule-dependent molecular motors. Tetratricopeptide repeat-containing (TTC) proteins are widely involved in protein complex formation. Nine of them are known to serve as components of the IFT or BBSome complexes. How many TTC proteins are cilia-related and how they function, however, remain unclear. Here we show that twenty TTC genes were upregulated by at least 2-fold during the differentiation of cultured mouse tracheal epithelial cells (MTECs) into multiciliated cells. Our systematic screen in zebrafish identified four novel TTC genes, ttc4, -9c, -36, and -39c, that are critical for cilia formation and motility. Accordingly, their zebrafish morphants displayed typical ciliopathy-related phenotypes, including curved body, abnormal otolith, hydrocephalus, and defective left-right patterning. The morphants of ttc4 and ttc25, a known cilia-related gene, additionally showed pronephric cyst formation. Immunoprecipitation indicated associations of TTC4, -9c, -25, -36, and -39c with components or entire complexes of IFT-A, IFT-B, or BBSome, implying their participations in IFT or IFT-related activities. Our results provide a global view for the relationship between TTC proteins and cilia.


Dynamically Evolving Surface Patterns through Light-Triggered Wrinkling Erasure.
For many applications, it is imperative that changes in polymer surface topography, especially periodic patterns, can be triggered on command by a well-defined remote signal. In this contribution, we report a light-induced cascade of changes in wrinkling wavelengths on thin polymer layers supported by an elastomeric substrate under tensile stress. Through the applied supramolecular design, the effect of varying the ratio of light-active and light-passive components can be easily assessed, and it is shown that both the cascade type as well as the rate of the progress of the dynamic light-induced changes can be tuned by this ratio as well as by the light intensity. Furthermore, for the reported phenomena to occur, nominally only every 20th polymer repeat unit needs to be occupied by a chromophore, which makes the conversion of the sub-nanometer photoisomerization reaction into 10 μm scale changes of periodic surface patterns extremely efficient.


Between life and death: strategies to reduce phototoxicity in super-resolution microscopy.
Super-resolution microscopy (SRM) enables non-invasive, molecule-specific imaging of the internal structure and dynamics of cells with sub-diffraction limit spatial resolution. One of its major limitations is the requirement for high-intensity illumination, generating considerable cellular phototoxicity. This factor considerably limits the capacity for live-cell observations, particularly for extended periods of time. Here, we give an overview of new developments in hardware, software and probe chemistry aiming to reduce phototoxicity. Additionally, we discuss how the choice of biological model and sample environment impacts the capacity for live-cell observations.


Mutations in BRCA1, BRCA2 and other breast and ovarian cancer susceptibility genes in Central and South American populations.
Breast cancer (BC) is the most common malignancy among women worldwide. A major advance in the understanding of the genetic etiology of BC was the discovery of BRCA1 and BRCA2 (BRCA1/2) genes, which are considered high-penetrance BC genes. In non-carriers of BRCA1/2 mutations, disease susceptibility may be explained of a small number of mutations in BRCA1/2 and a much higher proportion of mutations in ethnicity-specific moderate- and/or low-penetrance genes. In Central and South American populations, studied have focused on analyzing the distribution and prevalence of BRCA1/2 mutations and other susceptibility genes that are scarce in Latin America as compared to North America, Europe, Australia, and Israel. Thus, the aim of this review is to present the current state of knowledge regarding pathogenic BRCA variants and other BC susceptibility genes. We conducted a comprehensive review of 47 studies from 12 countries in Central and South America published between 2002 and 2017 reporting the prevalence and/or spectrum of mutations and pathogenic variants in BRCA1/2 and other BC susceptibility genes. The studies on BRCA1/2 mutations screened a total of 5956 individuals, and studies on susceptibility genes analyzed a combined sample size of 11,578 individuals. To date, a total of 190 different BRCA1/2 pathogenic mutations in Central and South American populations have been reported in the literature. Pathogenic mutations or variants that increase BC risk have been reported in the following genes or genomic regions: ATM, BARD1, CHECK2, FGFR2, GSTM1, MAP3K1, MTHFR, PALB2, RAD51, TOX3, TP53, XRCC1, and 2q35.


The emergence of maternal health as a political priority in Madhya Pradesh, India: a qualitative study.
Politics plays a critical role in agenda setting in health affairs; therefore, understanding the priorities of the political agenda in health is very important. The political priority for safe motherhood has been investigated at the national level in different countries. The objective of this study was to explore why and how maternal health became a political priority at sub-national level in the state of Madhya Pradesh in India. This study followed a qualitative design. Data were collected by carrying out interviews and review of documents. Semi-structured interviews were carried out with twenty respondents from four stakeholder groups: government officials, development partners, civil society and academics. Data analysis was performed using thematic analysis. The analysis was guided by Kingdon's multiple streams model. The emergence of maternal health as a political priority in Madhya Pradesh was the result of convergence in the developments in different streams: the development of problem definition, policy generation and political change. The factors which influenced this process were: emerging evidence of the high magnitude of maternal mortality, civil society's positioning of maternal mortality as a human rights violation, increasing media coverage, supportive policy environment and launch of the National Rural Health Mission (NRHM), the availability of effective policy solutions, India's aspiration of global leadership, international influence, maternal mortality becoming a hot debate topic and political transition at the national and state levels. Most of these factors first became important at national level which then cascaded to the state level. Currently, there is a supportive policy environment in the state for maternal health backed by greater political will and increased resources. However, malnutrition and population stabilization are the competing priorities which may push maternal health off the agenda. The influence of the events and factors evolving from international and national levels significantly contributed to the development of maternal health as a priority in Madhya Pradesh. This led to several opportunities in terms of policies, guidelines and programmes for improving maternal health. These efforts were successful to some extent in improving maternal health in the state but several implementation challenges still require special attention.


Novel FCN2 Variants and Haplotypes are Associated with Rheumatic Heart Disease.
Ficolins are pattern recognition molecules that are involved in innate immune defense. Ficonin-2 (FCN2) has a specific affinity for lipoteichoic acid present in the cell wall of Streptococcus pyogenes, an etiological agent for rheumatic heart disease (RHD). We have estimated FCN2 serum levels and analyzed the functional variants of FCN2 in 400 RHD patients, 617 healthy controls, and 581 individuals belonged to various ethnic populations, who are inhabited in various geographical regions of India. Our study revealed that the FCN2 -986A and +6359T alleles were the risk factors for RHD susceptibility (p = 0.0007 for -986G>A; p = 0.0004 for +6359C>T). The haplotype AGGT (p = 0.0024) was observed to be a risk factor for RHD susceptibility, and the haplotype GGAC (p = 0.002) was found to confer protection against RHD. The level of serum FCN2 was significantly higher in controls (p < 0.0001) and in controls with GGAC haplotypes (p < 0.0001). The frequency of the risk alleles -986A and +6359T was found to be more prevalent in Northern and North-Western (Indo-European) India. The protective GGAC haplotype was found more prevalent in Eastern (Tibeto-Burman) and South-Western (Dravidian) India. Alleles -986A and +6359T were in positive correlation with the prevalence of RHD (regression coefficient = 1.84 and 1.94, respectively), whereas GGAC haplotype was in negative correlation with prevalence of RHD (regression coefficient = -1.71). In conclusion, we found that low level of serum ficolin-2 is significantly associated with RHD. Further, FCN2 -986A and +6359T alleles and AGGT haplotype are associated with increased susceptibility to RHD, while GGAC haplotype is associated with moderate protection against RHD.


Bacterial Detection Rate and Surgical Outcome in Povidone-Iodine Irrigation After Nasolacrimal Duct Intubation.
In various ophthalmic surgical procedures, 0.25% povidone-iodine (PI) solution is used to prevent infections. This study examined the bacterial colonisation before and after endoscopic nasolacrimal duct intubation irrigated with PI solution and investigated the relationship between bacterial detection and surgical failure. A retrospective study at Saitama Medical University Hospital. We enrolled 113 patients (33 men, 80 women) who underwent lacrimal intubation surgery between April 2016 and December 2018. Preoperatively, the lacrimal pathways were washed with normal saline solution, which was subsequently cultured. The sites of obstruction in the lacrimal pathway were endoscopically determined, perforated, and intubated. The tubes were afterward either irrigated with 0.25% PI or normal saline every 2 weeks. After 8 weeks, a piece of the removed lacrimal tube was cultured. We characterized the pre- and postoperative bacterial cultures. We enrolled 54 and 52 patients in PI and saline groups (mean age: 69.1±9.2 and 73.2±7.2 years), respectively. In the PI group, Staphylococcus epidermidis was less frequently detected postoperatively than preoperatively. S. oralis and Candida parapsilosis were identified more often postoperatively (P=0.02, P=0.01, respectively). The PI group had significantly lower bacterial detection rate than the saline group (P=0.01). However, the surgical failure rates did not differ between groups. In surgical failure patients, the postoperative rate of Pseudomonas aeruginosa was very high (75%). PI reduces the bacterial detection rate but does not improve the surgical success rate. P. aeruginosa is associated with an increased risk of surgical failure.


Comparison of Human Surrogate Responses in Underbody Blast Loading Conditions.
Impact biomechanics research in occupant safety predominantly focuses on the effects of loads applied to human subjects during automotive collisions. Characterization of the biomechanical response under such loading conditions is an active and important area of investigation. However, critical knowledge gaps remain in our understanding of human biomechanical response and injury tolerance under vertically accelerated loading conditions experienced due to underbody blast (UBB) events. This knowledge gap is reflected in anthropomorphic test devices (ATDs) used to assess occupant safety. Experiments are needed to characterize biomechanical response under UBB relevant loading conditions. Matched pair experiments in which an existing ATD is evaluated in the same conditions as a post mortem human subject (PMHS) may be utilized to evaluate biofidelity and injury prediction capabilities, as well as ATD durability, under vertical loading. To characterize whole body response in the vertical direction, six whole body PMHS tests were completed under two vertical loading conditions. A series of 50th percentile hybrid III ATD tests were completed under the same conditions. Ability of the hybrid III to represent the PMHS response was evaluated using a standard evaluation metric. Tibial accelerations were comparable in both response shape and magnitude, while other sensor locations had large variations in response. Posttest inspection of the hybrid III revealed damage to the pelvis foam and skin, which resulted in large variations in pelvis response. This work provides an initial characterization of the response of the seated hybrid III ATD and PMHS under high rate vertical accelerative loading.


Comparative analysis of the MIR319a microRNA locus in Arabidopsis and related Brassicaceae.
MicroRNAs (miRNAs) are important regulators of gene expression in multicellular organisms. Yet, little is known about their molecular evolution. The 20- to 22-nt long miRNAs are processed in plants from foldbacks that are a few hundred base pairs in size. Often, these foldbacks are embedded in much larger precursor transcripts. To investigate functional constraints on sequence evolution of miRNA precursor genes, we have studied sequence variation in the precursor of miR319a, MIR319a, between species from the Brassicaceae. We compared the genomic context in Arabidopsis thaliana, Arabidopsis halleri, and Capsella rubella, using bacterial artificial chromosome clones, and analyzed precursor sequences obtained by polymerase chain reaction from 13 additional species. Phylogenetic shadowing identifies a conserved motif around the transcription start site, which we demonstrate to be functionally important. We further assessed the functionality of MIR319a orthologs from several Brassicaceae species in A. thaliana. The ortholog from kale (Brassica oleracea var. acephala) was found to be largely inactive, at least partially due to mutations in the miRNA itself, but experimental evidence suggests that loss of miR319a function is compensated by other members of the miR319 family. More broadly, we find that the foldback diverges less rapidly than the remainder of the primary transcript. To understand the molecular evolution of miRNA genes, investigations at different levels of phylogenetic divergence are required.


Community-Based Recreation Therapy and Mental Health Recovery: A Mixed-Media Participatory Action Research Study.
Personal recovery is an individualized process through which people develop a positive identity and live a meaningful life, with symptoms of mental illness. Few studies have explored the role of recreation therapy in the recovery process from the perspectives of individuals with lived experience of mental illness. To understand how community-based recreation therapy can support mental health recovery, from the perspectives of people diagnosed with mental illness, and to guide the development, delivery and evaluation of recovery-oriented mental health services. Guided by the principles of participatory action research (PAR) and photovoice, six participant researchers (PRs) generated arts-based media and narrative data in response to the research question: How can therapeutic recreation, in a community mental health center, support the recovery of individuals diagnosed with mental illness? The research group analyzed the qualitative data through a participatory data analysis process. The PRs produced and analyzed 24 pieces of arts-based media and 5 hours of transcribed narrative data describing their artworks' relationship to therapeutic recreation and recovery. The PRs identified seven salient themes through the participatory data analysis process: providing a safe place, promoting hope, finding balance, developing self-wisdom, increasing enjoyment, building confidence, and encouraging self-determination. Service recipients' unique preferences and perspectives must be integral to service development to deliver therapeutic recreation interventions that are truly recovery-oriented. The research findings will be used to guide the delivery of innovative, collaborative, person-centered programming in community-based mental health settings.


Inactivation of Transcriptional Regulators during Within-Household Evolution of Escherichia coli.
We analyzed the within-household evolution of two household-associated Escherichia coli strains from pandemic clonal group ST131-H30, using isolates recovered from five individuals within two families, each of which had a distinct strain. Family 1's strain was represented by a urine isolate from the index patient (older sister) with recurrent cystitis and a blood isolate from her younger sister with fatal urosepsis. Family 2's strain was represented by a urine isolate from the index patient (father) with pyelonephritis and renal abscesses, blood and kidney drainage isolates from the daughter with emphysematous pyelonephritis, and urine and fecal isolates from the mother with cystitis. Collectively, the several variants of each family's strain had accumulated a total of 8 (family 1) and 39 (family 2) point mutations; no two isolates were identical. Of the 47 total mutations, 36 resulted in amino acid changes or truncation of coded proteins. Fourteen such mutations (39%) targeted genes encoding transcriptional regulators, and 9 (25%) involved DNA-binding transcription factors (TFs), which significantly exceeded the relative contribution of TF genes to the isolates' genomes (∼6%). At least one-half of the transcriptional regulator mutations were inactivating, based on phenotypic and/or transcriptional analysis. In particular, inactivating mutations in the global regulator LrhA (repressor of type 1 fimbriae and flagella) occurred in the blood isolates from both households and increased the virulence of E. coli strains in a murine sepsis model. The results indicate that E. coli undergoes adaptive evolution between and/or within hosts, generating subpopulations with distinctive phenotypes and virulence potential.IMPORTANCE The clonal evolution of bacterial strains associated with interhost transmission is poorly understood. We characterized the genome sequences of clonal descendants of two Escherichia coli strains, recovered at different time points from multiple individuals within two households who had different types of urinary tract infection. We found evidence that the E. coli strains underwent extensive mutational diversification between and within these individuals, driven disproportionately by inactivation of transcriptional regulators. In urosepsis isolates, the mutations observed in the global regulator LrhA increased bacterial virulence in a murine sepsis model. Our findings help in understanding the adaptive dynamics and strategies of E. coli during short-term natural evolution.


Adolescents in wartime US military families: a developmental perspective on challenges and resources.
Adolescents in wartime US military families are a unique group of young people who are experiencing the usual milestones of adolescent development, including establishing their identities and becoming autonomous, while they face the challenges of military life such as multiple frequent moves, relocation, and parent deployment to combat settings. This paper reviews research on adolescents in wartime US military families, within the context of adolescent development, to identify their behavioral, emotional and academic risk status, and challenges and resources. Recommendations for future research and interventions to foster the healthy development of these adolescents are also provided.


Purification and characterization of an extracellular heme-binding protein, HasA, involved in heme iron acquisition.
Many bacterial hemoproteins involved in heme acquisition have been isolated recently, comprising outer membrane receptors and extracellular heme-binding protein. The mechanisms by which these proteins extract heme have not been described up to now. One such protein, HasA, which can bind free heme as well as capture it from hemoglobin, is secreted by the Gram-negative bacteria Serratia marcescens under iron deficiency conditions. The fact that HasA does not present sequence similarities with other known hemoproteins suggests that it possesses a new type of heme binding site. This work describes the main physicochemical properties of HasA, essential for understanding its function. HasA is a monomer of 19 kDa that binds one b heme per molecule with high affinity. The electron paramagnetic resonance spectra indicate that the heme iron is in a low-spin ferric state and that the two iron axial ligands are His and His-. The low oxidation-reduction potential value (-550 mV vs standard hydrogen electrode) of the heme bound to HasA suggests that heme could be exposed to the solvent. According to circular dichroism data, the binding of heme does not seem to modify the conformation of HasA.


Hyperbaric oxygen: B-level evidence in mild traumatic brain injury clinical trials.
First, to demonstrate that B-level evidence exists for the use of hyperbaric oxygen therapy (HBOT) as an effective treatment in mild to moderate traumatic brain injury/persistent postconcussion syndrome (mTBI/PPCS). Second, to alert readers and researchers that currently used pressurized air controls (≥21% O2, >1.0 ATA) are therapeutically active and cannot be utilized as sham controls without further validation. Review of published, peer-reviewed articles of HBOT prospective and controlled clinical trials of mTBI/PPCS symptoms. Published results demonstrate that HBOT is effective in the treatment of mTBI/PPCS symptoms. Doses of oxygen that are applied at ≥21% O2 and at pressures of >1.0 ATA produce improvements from baseline measures. Some of the recently published clinical trials are mischaracterized as sham-controlled clinical trials (i.e., sham = 21% O2/1.2-1.3 ATA), but are best characterized as dose-varying (variation in oxygen concentration, pressure applied, or both) clinical trials. Hyperbaric oxygen and hyperbaric air have demonstrated therapeutic effects on mTBI/PPCS symptoms and can alleviate posttraumatic stress disorder symptoms secondary to a brain injury in 5 out of 5 peer-reviewed clinical trials. The current use of pressurized air (1.2-1.3 ATA) as a placebo or sham in clinical trials biases the results due to biological activity that favors healing.


Targeting ferroptosis in hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is a common malignant tumor with complex survival mechanism and drug resistance, resulting in cancer-related high mortality in the world. Ferroptosis represents a form of regulated cell death, typically distinguished by iron-dependent lipid peroxidation. Cancer cells often employ antioxidant defenses to evade the harmful effects of excess iron. Recent research has proposed that directing interventions towards ferroptosis could serve as an effective strategy in curbing the proliferation and invasion of HCC. Immunotherapy has made some preliminary progress in the remodeling of immune microenvironment, but it has not completely inhibited HCC growth, invasion and drug resistance. Furthermore, ferroptosis is widely observed in the formation of immune microenvironment of HCC and mediates the response of many targeted drugs and immunotherapy. Clarifying the role of ferroptosis in these complex processes is expected to provide a new prospect for HCC treatment. In this review, we outline the mechanisms by which HCC develops invasiveness and drug resistance by evading iron-dependent death, and paint a comprehensive landscape of ferroptosis in different cell types in the HCC immune microenvironment.


Sex differences in the reciprocal relationship between glycemic control and depressive symptoms among older adults with diabetes: Using a nationwide population-based sample.
As life expectancy increases worldwide, the prevalence and the disease burden of diabetes in older adults are also increasing. This study aimed to examine sex differences in the reciprocal relationship between glycemic control and depressive symptoms among older adults with diabetes by using longitudinal data at two timepoints. Wave 1 (W1, 2016-2017) and wave 2 (W2, 2018-2019) data from the Korean Frailty and Aging Cohort Study were used. Finally, 416 older adults with diabetes who satisfied the inclusion criteria were analyzed (215 males, 201 females). The reciprocal relationship between depressive symptoms and glycosylated hemoglobin A1c (HbA1c) levels was examined using a cross-lagged panel model. HbA1c levels and depressive symptoms at earlier time points were the most significant factors contributing to HbA1c levels and depressive symptoms at later timepoints. The relationship between HbA1c level and depressive symptoms differed according to sex. The cross-path from depressive symptoms (W1) to HbA1c levels (W2) was positively statistically significant in males (β = 0.18, SD = 0.05, P = 0.001). The path from HbA1c levels (W1) to depressive symptoms (W2) was positively statistically significant in females (β = 0.12, SD = 0.06, P = 0.032). Early management of blood glucose levels and depressive symptoms is important in older adults with diabetes. Moreover, glycemic control through social activities in the community could be effective in relieving depressive symptoms in older females with diabetes, and managing depressive symptoms and glucose levels together could be effective in glycemic control in older males with diabetes. Geriatr Gerontol Int 2023; ••: ••-••.


[Value of informed consent in tympanoplasty and FESS procedures].
More than one third of all lawsuits against surgeons include statements of insufficient or lacking preoperative informed consent which is mandatory in the german jurisprudence. Almost nothing is known about the postoperative patient's memory of risk factors which were explained prior to typical surgical procedures in ENT. The objectives of this prospective study in 201 patients were to elucidate the actual clinical routine of obtaining informed consent for tympanoplasty (n=105) and FESS procedures (n=89), collect information on active and passive recall 6 weeks, 6 months and 1 year post-operatively, and to investigate whether patient age, sex, education, the time from obtaining consent to interview, recurrent vs. non-recurrent procedures, would influence the patient's recall capabilities. In clinical routine, obtaining informed consent included 5 main items for tympanoplasty and 5 for FESS procedures made by the physician. Of the patients, 18.9% recalled actively and 65.3% passively. They named 1.5 items on average, with "deafness" as the leading complication in tympanoplasty and "amaurosis" in FESS procedures. Patient's memory was depending on the time from obtaining consent to interview and education exclusively. In conclusion, the quantity of patient/physician interaction does not guarantee an increased effect on patient's recall. Improving patient interaction may reduce the probability of imminent accusations.


Serum antibody responses to individual viral polypeptides in human rotavirus infections.
A radioimmunoprecipitation assay (RIPA) was used to study the serum antibody responses to individual polypeptides that developed after infection with viruses from human rotavirus subgroups I and II. Paired sera from eight children (1 to 8.5 years of age) were used in the study. Although all of the eight acute sera were negative by the complement fixation test, four of them were positive by RIPA, indicating a previous infection by rotavirus. A significant difference in the number of polypeptides immunoprecipitated was seen among the convalescent sera. The number of polypeptides immunoprecipitated was found to be related to previous infection experience. At most, ten different polypeptides were immunoprecipitated: seven structural polypeptides VP1 to VP7 and three non-structural polypeptides, NS1, NS2 and NS3. No sera immunoprecipitated VP8 or VP9. Acute sera positive by RIPA immunoprecipitated up to five polypeptides, VP1, VP2, VP3, VP4 and VP6. One of the non-structural proteins (NS2) was found to be particularly immunogenic, since antibodies to this polypeptide were detected in several convalescent sera. Among the structural proteins VP2 and VP6 were found to be the two immunodominant polypeptides which were recognized by all convalescent sera. Only three convalescent sera immunoprecipitated VP7, the major type-specific antigen responsible for inducing neutralizing antibodies. Three of four originally seronegative children with no reactivity in the convalescent sera to VP7 developed neutralizing antibodies to a single serotype. One child developed antibodies to two serotypes.


Mss51p promotes mitochondrial Cox1p synthesis and interacts with newly synthesized Cox1p.
The post-transcriptional role of Mss51p in mitochondrial gene expression is of great interest since MSS51 mutations suppress the respiratory defect caused by shy1 mutations. SHY1 is a Saccharomyces cerevisiae homolog of human SURF1, which when mutated causes a cytochrome oxidase assembly defect. We found that MSS51 is required for expression of the mitochondrial reporter gene ARG8(m) when it is inserted at the COX1 locus, but not when it is at COX2 or COX3. Unlike the COX1 mRNA-specific translational activator PET309, MSS51 has at least two targets in COX1 mRNA. MSS51 acts in the untranslated regions of the COX1 mRNA, since it was required to synthesize Arg8p when ARG8(m) completely replaced the COX1 codons. MSS51 also acts on a target specified by the COX1 coding region, since it was required to translate either COX1 or COX1:: ARG8(m) coding sequences from an ectopic COX2 locus. Mss51p was found to interact physically with newly synthesized Cox1p, suggesting that it could coordinate Cox1p synthesis with insertion into the inner membrane or cytochrome oxidase assembly.


Human complement activation and anaphylatoxins generation induced by snake venom toxins from Bothrops genus.
Snake venoms are a complex mixture of components, which have a wide range of actions both on prey and human victims. The genus Bothrops causes the vast majority of snakebites in Central and South America, being responsible for 80% of snake envenomations in Brazil. Envenomations are characterized by prominent local effects, including oedema, haemorrhage and necrosis, which can lead to permanent disability. Systemic manifestations such as haemorrhage, coagulopathy, shock and acute renal failure may also occur. In the present study we have investigated the action of venoms from 19 species of snakes from the genus Bothrops, occurring in Brazil, on the complement system in in vitro studies. All venoms were able to activate the classical complement pathway, in the absence of sensitizing antibody. This activation was in part associated with the cleavage of C1-Inhibitor by proteases present in these venoms, which disrupts complement activation control. No modification of the membrane bound complement regulators, such as DAF, CR1 and CD59 was detected, after treatment of human erythrocytes with the snake venoms. Some of the Bothrops venoms were also able to activate alternative and lectin pathways, as measured in haemolytic and ELISA assays. C3a, C4a and C5a were generated in sera treated with the venoms, not only through C-activation, but also by the direct cleavage of complement components, as determined using purified C3 and C4. Metallo- and/or serine-protease inhibitors prevented cleavage of C3 and C4. These results suggest that Bothrops venoms can activate the complement system, generating a large amount of anaphylatoxins, which may play an important role in the inflammatory process presented in humans after snake envenomations, and they may also assist, due to their vasodilatory effects, to enhance the spreading of other venom components.


Topological phases of monolayer and bilayer depleted Lieb lattices.
Existence of nontrivial topological phases in a tight binding Haldane-like model on the depleted Lieb lattice is reported. This two-band model is formulated by considering the nearest-neighbor, next-nearest-neighbor and next-next-nearest-neighbor hopping terms along with complex phase which breaks the time reversal symmetry of this semi-metallic system. Topological feature of this model is studied along with the presence of sublattice symmetry breaking staggered onsite energy. Combined effect of these two broken symmetries is found crucial to lead an additional transition between nontrivial to trivial phases. System exhibits two types of phase transitions, say, between two nontrivial phases and nontrivial to trivial phases. Nonzero Chern numbers, existence of Hall plateau and symmetry protected edge states confirm the presence of the nontrivial phases. This two-band system hosts four different types of phases where two are topological. Additionally topological properties of stacked bilayer of the depleted Lieb lattices is also studied with similar Haldane-like Hamiltonian. This four-band system is found to host Chern insulating phases, with higher values of Chern numbers supported by ingap edge states.&#xD.


Evidence against direct involvement of cyclic GMP or cyclic AMP in bacterial chemotactic signaling.
Defects in phosphotransferase chemotaxis in cya and cpd mutants previously cited as evidence of a cyclic GMP or cyclic AMP intermediate in signal transduction were not reproduced in a study of chemotaxis in Escherichia coli and Salmonella typhimurium. In cya mutants, which lack adenylate cyclase, the addition of cyclic AMP was required for synthesis of proteins that were necessary for phosphotransferase transport and chemotaxis. However, the induced cells retained normal phosphotransferase chemotaxis after cyclic AMP was removed. Phosphotransferase chemotaxis was normal in a cpd mutant of S. typhimurium that has elevated levels of cyclic GMP and cyclic AMP. S. typhimurium crr mutants are deficient in enzyme III glucose, which is a component of the glucose transport system, and a regulator of adenylate cyclase. After preincubation with cyclic AMP, the crr mutants were deficient in enzyme II glucose-mediated transport and chemotaxis, but other chemotactic responses were normal. It is concluded that cyclic GMP does not determine the frequency of tumbling and is probably not a component of the transduction pathway. The only known role of cyclic AMP is in the synthesis of some proteins that are subject to catabolite repression.


Impaired glucose tolerance is associated with increased serum concentrations of interleukin 6 and co-regulated acute-phase proteins but not TNF-alpha or its receptors.
A population-based sample was studied to define immune abnormalities in individuals at risk of Type II (non-insulin-dependent) diabetes mellitus because of impaired glucose tolerance. A total of 1653 individuals aged 55 to 74 years participated in a population based survey in Southern Germany (KORA Survey 2000). Those without a history of diabetes were subjected to an OGTT. Randomly selected subjects with IGT ( n=80) were compared with non-diabetic control subjects ( n=77) and patients with Type II diabetes ( n=152) of the same population-based sample after matching for age and sex. Immune parameters were analysed in serum with rigidly evaluated ELISA. Serum pro-inflammatory cytokine interleukin 6 (IL-6) concentrations were higher in subjects with IGT and Type II diabetes than in the control subjects (median 1.8 and 2.5 vs 0.8 pg/ml, p<0.0001). Soluble IL-6 receptors potentiate IL-6 bioactivity and their concentrations were mildly increased in Type II diabetes ( p<0.05). These immune changes seem relevant because IL-6 dependent acute-phase proteins C-reactive protein, serum amyloid A protein and fibrinogen were also increased in IGT and Type II diabetes. Circulating concentrations of TNF-alpha and its two receptors sTNF-R60 and sTNF-R80 were not increased in IGT subjects compared with the control subjects. Our study shows systemic up-regulation of selected inflammatory mediators in patients with Type II diabetes and IGT. The pattern observed is non-random and fits with an IL-6 associated rather than TNF-alpha associated response.