While the effects of benzo[c]phenanthridine alkaloids (QBA), known mainly as sanguinarine and chelerythrine, on the inhibition of some kinds of cancer cell proliferation have been established, the effect on oral squamous cell is not known. Here, the antitumor activity of sanguinarine was demonstrated using in vitro assay systems in SAS, a human oral squamous cell carcinoma (OSCC) cell line. The anti-proliferative and -invasive effects were confirmed with IC₅₀ values in the concentration range of 0.75-1.0 μM by MTT assay and invasive assay, respectively. Sanguinarine was also able to suppress cell anchorage-independent growth, whereas it did not affect the cells' adhering capabilities. Finally, sanguinarine induced apoptotic cell death by activating caspase and altering the Bcl-2/Bax ratio. Taken together, these results indicate that sanguinarine is a potential inhibitor of tumorigenesis and suggest that it may be valuable in the development of new anticancer drugs for the treatment of OSCC.

zeta-Globin chain expression in carriers of a number of deletional alpha-thalassemias is investigated by radioimmunoassay. In a few cases, zeta-globin mRNAs are also studied. zeta-Globin chains are detected in (--SEA/), (--MED/), and (--SPAN/) deletions, but not in six other deletional mutations. These results suggest that the DNA element capable of suppressing zeta-globin expression in adult erythroid cells is present within the (--SPAN/) deletion, while the DNA fragment between the 5' breakpoints of the (--SA/) and the (--SEA/) deletions may contain sequences necessary for augmenting zeta-globin expression in adult erythroid cells. Furthermore, zeta-globin chains are shown by an immunocytologic technique to be present in all circulating erythrocytes in carriers of the (--SEA/) and (--MED/) deletions. This simple immunocytologic test is highly sensitive and specific to detect adult carriers of either the (--SEA/) or (--MED/) deletions, and can be used for the detection of couples at risk of pregnancies involving fetuses with homozygous alpha-thalassemia.

Primary aldosteronism (PA) is the most common secondary cause of hypertension that has recently been implicated in alterations of the immune system and progression of cardiovascular disease.

OBJECTIVE

To study the cytokines transforming growth factor beta1 (TGF-beta1), tumor necrosis factor alpha (TNF-alpha), and interleukin 10 (IL-10) in patients with PA and essential hypertensives (EH) and evaluate its association with the renin-angiotensin-aldosterone system.

METHODS

We studied 26 PA and 52 EH patients as controls, adjusted by their blood pressure, body mass index, age, and gender. In both groups, PA and EH, we measured serum aldosterone (SA), plasma renin activity (PRA), and cytokines TGF- beta1, TNF-alpha, and IL-10. In addition, 17 PA patients were treated for 6 months with spironolactone, a mineralocorticoid receptor (MR) antagonist.

RESULTS

PA patients had lower levels of TGF-beta1 (17.6+/-4.1 vs 34.5+/-20.5 pg/ml, p<0.001) and TNF-alpha (17.0+/-4.4 vs 35.6+/-21.7 pg/ml, p<0.001) and similar IL-10 levels (99.7+/-18.7 vs 89.4+/-49.5 pg/ml, p: ns), as compared with EH controls. TGF-beta1 and TNF-alpha levels showed a remarkable correlation with SA/PRA ratio in the total group (PA+EH). The treatment of PA patients with spironolactone increased the TGF-beta1 levels (18.3+/-5.9 to 28.4+/-6.3 pg/ml, p<0.001), while TNF-alpha, and IL-10 remained unchanged.

CONCLUSIONS

Our results showed that PA patients have lower TGF-beta1 and TNF-alpha cytokine serum levels than EH. TGF-beta1 levels were restored with spironolactone, showing a MR-dependent regulation. In this way, the chronic aldosterone excess modifies the TGF-beta1 levels, which could produce an imbalance in the immune system homeostasis that may promote an early proinflammatory cardiovascular phenotype.

Limited information is available on the identity of antigens targeted by antibodies present in cerebrospinal fluid (CSF) of patients with clinically isolated syndrome (CIS). The aim of this study was to identify novel antigens for CIS and investigate their prognostic potential to predict conversion to multiple sclerosis (MS). We applied serological antigen selection (SAS) to identify antigens interacting with antibodies present in the pooled CSF from four CIS patients, who developed MS. Antibody reactivity towards CIS antigens identified by SAS was tested in CSF and serum from patients with CIS (n = 123/n = 108), MS (n = 65/n = 44), and other (inflammatory) neurological diseases (n = 75/n = 38) as well as in healthy control sera (n = 44). Using SAS, a panel of six novel CIS candidate antigens was identified. CSF antibody reactivity was detected in both CIS and relapsing-remitting (RR) MS. Serum reactivity was significantly increased in CIS and RR-MS as compared with controls (p = 0.03). For two antigens, the frequency of antibody-positive patients was higher in CIS patients who converted to MS as compared with CIS patients without conversion. We identified novel CIS antigens to which antibody reactivity was primarily detected in CIS and RR-MS as compared to controls. Possible prognostic potential could be demonstrated for two antigens.

As a widespread pollutant in the environment, cadmium (Cd) would be accumulated in leaves and cause phytotoxic effect on plants. Salicylic acid (SA), a natural signal molecule, plays an important role in eliciting specific responses to biotic and abiotic stresses. In our case, the effect of SA on Cd-induced photochemical damage and cell death in Arabidopsis was studied. The results illustrated that Cd could cause a series of physiological events such as chloroplast structure change (e.g. irregular mesophyll cell as well as ultrastructure change), reactive oxygen species (ROS) production and cell death. Furthermore, chlorophyll fluorescence parameters (F(v)/F(m), qN and ETR) showed a rapid decrease in wild-type (WT) Arabidopsis after treatment with 50 μM CdCl(2), identical with the change in chlorophyll delayed fluorescence (DF) intensity. The changes of these parameters showed the damage of Cd toxicity to photosynthetic apparatus. We found that cell death might be autophagic cell death, which might be caused by Cd toxicity induced oxidative stress just like photosynthetic damage. The NahG plants with lower SA accumulation level showed more sensitivity to Cd toxicity, although they exhibited a decrease both in chlorophyll fluorescence parameters and DF intensity. Exogenously SA prevented the Cd-induced photochemical efficiency decrease and mitigated Cd toxicity. Additionally, SA pretreatment could alleviate Cd-induced ROS overproduction. In conclusion, our results suggested that SA could prevent Cd-induced photosynthetic damage and cell death, which might be due to the inhibition of ROS overproduction.

Volatile, low-molecular weight terpenoids have been implicated in plant defenses, but their direct role in resistance against microbial pathogens is not clearly defined. We have examined a possible role of terpenoid metabolism in the induced defense of Arabidopsis thaliana plants against leaf infection with the bacterial pathogen Pseudomonas syringae. Inoculation of plants with virulent or avirulent P. syringae strains induces the emission of the terpenoids (E,E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene (TMTT), beta-ionone and alpha-farnesene. While the most abundant volatile, the C16-homoterpene TMTT, is produced relatively early in compatible and incompatible interactions, emission of both beta-ionone and alpha-farnesene only increases in later stages of the compatible interaction. Pathogen-induced synthesis of TMTT is controlled through jasmonic acid (JA)-dependent signaling but is independent of a functional salicylic acid (SA) pathway. We have identified Arabidopsis T-DNA insertion lines with defects in the terpene synthase gene TPS4, which is expressed in response to P. syringae inoculation. The tps4 knockout mutant completely lacks induced emission of TMTT but is capable of beta-ionone and alpha-farnesene production, demonstrating that TPS4 is specifically involved in TMTT formation. The tps4 plants display at least wild type-like resistance against P. syringae, indicating that TMTT per se does not protect against the bacterial pathogen in Arabidopsis leaves. Similarly, the ability to mount SA-dependent defenses and systemic acquired resistance (SAR) is barely affected in tps4, which excludes a signaling function of TMTT during SAR. Besides P. syringae challenge, intoxication of Arabidopsis leaves with copper sulfate, a treatment that strongly activates JA biosynthesis, triggers production of TMTT, beta-ionone, and alpha-farnesene. Taken together, our data suggest that induced TMTT production in Arabidopsis is a by-product of activated JA signaling, rather than an effective defense response that contributes to resistance against P. syringae.

Plant fatty acid alpha-dioxygenases (DOXs) catalyze the stereospecific conversion of fatty acids into the corresponding (R)-2-hydroperoxy fatty acids. In several plant species the corresponding gene was shown to be induced by pathogen infection, herbivore attack and environmental stresses. The precise signaling pathway accountable for the induction remains unidentified. In the present study, the effects of bacterial infection, oxidative- and heavy metal-stresses, and plant signaling molecules such as jasmonate, salicylic acid (SA), and ethylene (ET) on expression of a fatty acid alpha-DOX (OsDOX) gene in rice seedlings were investigated. The rice blight bacteria, Xanthomonas oryzae, elicited the accumulation of OsDOX transcripts in the leaves in both the incompatible and compatible interactions. Treating the seedling with CuSO4 also significantly enhanced the OsDOX expression. The degree of induction was shown to be mostly parallel to the level of endogenous jasmonic acid (JA) in the leaves. In contrast, SA was little effective and ET down-regulated not only the OsDOX expression but also the endogenous level of JA in rice seedlings. These results suggested that the OsDOX gene expression by a variety of stress-related stimuli was activated through jasmonate signaling and was negatively regulated by ET.

Nonconventional biomacromolecular luminogens have attracted extensive interest due to their fundamental importance and potential applications in diverse areas. To explore novel luminogens and, moreover, to gain deeper insights into their emission mechanism, we study the emission behaviors of sodium alginate (SA), a natural anionic polysaccharide composed of mannuronic (M) and guluronic acids (G). We find that the luminescence from aqueous SA solutions exhibits distinct concentration enhanced emission and aggregation-induced emission (AIE) characteristics. Meanwhile, the ratio of M/G also matters. Rheological measurements reveal the distinct regimes of the solutions, which are consistent with the observed emission, indicative of strong association between the chain entanglement and emission. Moreover, we observe persistent room temperature phosphorescence (RTP) in the amorphous SA solids, which is a rare case even in pure organic aromatic luminogens. Such unique emission can be remarkably enhanced via coordination with Caaviors can be well rationalized by the clustering-triggered emission (CTE) mechanism. Namely, the emission is caused by the electron cloud overlap due to the clustering of oxygen atoms and carboxylate units, together with conformation rigidification. Owing to its biocompatibility, intrinsic emission, and, moreover, persistent RTP, SA shows great potential for anticounterfeiting, encryption, intracellular imaging, and so on.

Background: Liver cancer stem cells (LCSCs) are responsible for the initiation, progression and chemoresistance of liver cancer. However, no agent targeting LCSC is available in the clinic to date. Here, we investigated the effects of targeting protein arginine methyltransferase 5 (PRMT5), an epigenetic regulator, on LCSCs and HCC using a novel PRMT5 inhibitor DW14800. Methods: Tumor spheroid formation culture was used to enrich LCSCs and assess their self-renewal capability. Human alpha-1-antitrypsin (A1AT) ELISA, acetylated low-density lipoprotein (ac-LDL) uptake, periodic acid-Schiff (PAS) reactions and senescence associated β-galactosidase (SA-β-gal) activity assays were performed to examine the differentiation status of HCC cells. The effects of DW14800 on HCC malignancy were assessed in HCC cell lines and on an HCC xenograft model in mice. Chromatin immunoprecipitation was applied to clarify the transcriptional regulation of HNF4α by PRMT5-mediated Histone H4 arginine-3 symmetrical dimethylation (H4R3me2s). Results: Quantitative real-time PCR revealed that the expression of PRMT5 was upregulated in LCSCs. DW14800 specifically decreased the symmetrical dimethylation of arginine residues in HCC cells. Treatment of DW14800 suppressed the self-renewal capacity of LCSCs while re-establishing hepatocyte-specific characteristics in HCC cells. DW14800 displayed antitumor effects in HCC cells in vitro and in xenograft HCC in vivo. Importantly, ChIP assay showed that PRMT5 and H4R3me2s bound to the promoter region of HNF4α gene, and DW14800 increased the expression of HNF4α via reducing the H4R3me2s levels and enhancing the transcription of HNF4α. Conclusions: Our data revealed the significance of targeting PRMT5 activity in LCSC elimination and HCC differentiation, and proposed that DW14800 may represent a promising therapeutic agent for HCC in the clinic.

Antibiotic use has been limited in U.S. swine production. Therefore, the objective was to determine whether supplementing l-glutamine at cost-effective levels can replace dietary antibiotics to improve piglet welfare and productivity following weaning and transport. Based on previous research, we hypothesized that withholding dietary antibiotics would negatively affect pigs while diet supplementation with 0.20% l-glutamine (GLN) would have similar effects on pig performance and health as antibiotics. Mixed sex piglets (N = 480; 5.62 ± 0.06 kg BW) were weaned (18.4 ± 0.2 d of age) and transported for 12 h in central Indiana, for 2 replicates, during the summer of 2016 and the spring of 2017. Pigs were blocked by BW and allotted to 1 of 3 dietary treatments (n = 10 pens/dietary treatment/replicate [8 pigs/pen]); antibiotics (A; chlortetracycline [441 ppm] + tiamulin [38.6 ppm]), no antibiotics (NA), or GLN fed for 14 d. On days 15 to 34, pigs were provided common antibiotic-free diets in 2 phases. Data were analyzed using PROC MIXED in SAS 9.4. Day 14 BW and days 0 to 14 ADG were greater (P = 0.01) for A (5.6% and 18.5%, respectively) and GLN pigs (3.8% and 11.4%, respectively) compared with NA pigs, with no differences between A and GLN pigs. Days 0 to 14 ADFI increased for A (P < 0.04; 9.3%) compared with NA pigs; however, no differences were detected when comparing GLN with A and NA pigs. Once dietary treatments ceased, no differences (P > 0.05) in productivity between dietary treatments were detected. On day 13, plasma tumor necrosis factor alpha (TNF-α) was reduced (P = 0.02) in A (36.7 ± 6.9 pg/mL) and GLN pigs (40.9 ± 6.9 pg/mL) vs. NA pigs (63.2 ± 6.9 pg/mL). Aggressive behavior tended to be reduced overall (P = 0.09; 26.4%) in GLN compared with A pigs, but no differences were observed between A and GLN vs. NA pigs. Huddling, active, and eating/drinking behaviors were increased overall (P < 0.02; 179%, 37%, and 29%, respectively) in the spring replicate compared with the summer replicate. When hot carcass weight (HCW) was used as a covariate, loin depth and lean percentage were increased (P = 0.01; 4.0% and 1.1%, respectively) during the spring replicate compared with the summer replicate. In conclusion, GLN supplementation improved pig performance and health after weaning and transport similarly to A across replicates; however, the positive effects of A and GLN were diminished when dietary treatments ceased.

We fabricate ultrasmall phosphorene quantum dots (PQDs) with an average size of 2.6 ± 0.9 nm using a liquid exfoliation method involving ultrasound probe sonication followed by bath sonication. By coupling the as-prepared PQDs with microfiber evanescent light field, the PQD-based saturable absorber (SA) device exhibits ultrafast nonlinear saturable absorption property, with an optical modulation depth of 8.1% at the telecommunication band. With the integration of the all-fiber PQD-based SA, a continuous-wave passively mode-locked erbium-doped (Er-doped) laser cavity delivers stable, self-starting pulses with a pulse duration of 0.88 ps and at the cavity repetition rate of 5.47 MHz. Our results contribute to the growing body of work studying the nonlinear optical properties of ultrasmall PQDs that present new opportunities of this two-dimensional (2D) nanomaterial for future ultrafast photonic technologies.

OBJECTIVE

To evaluate the clinical performance of Scotchbond Universal (3M Oral Care) and Prime & Bond Elect (Dentsply Sirona) in the restoration of noncarious cervical lesions (NCCLs).

METHODS

This was a randomized controlled clinical trial involving 63 subjects. Two hundred and three NCCLs were restored using Scotchbond Universal and Prime & Bond Elect using both an etch-and-rinse and a self-etch technique. Lesions were notch-shaped NCCLs, and the restorations were placed without any mechanical retention. Restorations were finished immediately after placement and scored with regard to retention, marginal discoloration, marginal adaptation, and secondary caries. Similar assessment of the restorations was performed 18 months after placement. Logistic regression was performed for each outcome separately with a compound symmetric variance-covariance structure assumed to consider a correlation of restorations within subjects. All analyses were conducted using SAS version 9.4 (SAS Inc).

RESULTS

One hundred and fifty-eight teeth (77.8% of the restorations placed) in 46 subjects (73% of subjects enrolled) were available for the 18-month follow-up. A statistically significant difference was reached only for the comparison Scotchbond Universal/self-etch (SU_SE) and Prime & Bond Elect/etch-and-rinse (PBE_E&R) groups ( p=0.01), where a restoration with SU_SE was 66% less likely to maintain a score of Alpha for marginal discoloration than a restoration performed with PBE_E&R.

CONCLUSIONS

Scotchbond Universal and Prime & Bond Elect presented acceptable clinical performance after 18 months of clinical service. However, Scotchbond Universal, when applied with a self-etch approach, did demonstrate a relatively high level of marginal discoloration when compared to the other groups.

A cell-type saturable absorber has been demonstrated by filling the single mode photonic crystal fiber (SMPCF) with tungsten disulfide (WS2) nanosheets. The modulation depth, saturable intensity, and non-saturable loss of this SA are measured to be 3.53%, 159 MW/cm(2) and 23.2%, respectively. Based on this SA, a passively mode-locked EDF laser has been achieved with pulse duration of 808 fs and repetition rate of 19.57 MHz, and signal-noise-ratio (SNR) of 60.5 dB. Our results demonstrate that the cell-type WS2 nanosheets SA can serve as a good candidate for short-pulse mode locker.

Acute osteoarticular infection (OAI) is a potentially severe disease. The aim of this study was to evaluate the etiology, clinical characteristics and therapeutic approach of OAI in children in Spain.

Medical records from children <14 years with OAI from 25 hospitals between 2008 and 2012 were reviewed. Confirmed osteomyelitis (OM) and septic arthritis (SA) required a positive bacterial isolate; otherwise, they were considered probable. Probable SA with <40,000 cells/mm in joint fluid was not included.

A total of 641 children were evaluated. Two hundred and ninety-nine cases (46%) were OM, 232 (36%) SA, 77 (12%) osteoarthritis and 33 (5%) spondylodiscitis. Children with OM were older (63 vs. 43 months for SA; P < 0.001). Magnetic resonance imaging and bone scintigraphy had the highest yield for OM diagnosis (94%). Arthrocentesis was performed in 96% of SA. A microorganism was isolated in 246 patients (38%: 33% OM vs. 55% SA; P < 0.001): Staphylococcus aureus was the most common (63%), followed by Kingella kingae (15%) and Streptococcus pyogenes (9%). Ninety-five percent of children initially received IV antibiotics, mostly cefotaxime + cloxacillin (60%) or cloxacillin (40%). Total treatment duration was 38 (±31) days for OM and 28 (±16) days for SA (P < 0.0001). Twenty percent of children with OM (46% because of complications) and 53% with SA (95% initial arthrotomy) underwent surgery. Patients with SA were compared according to initial arthrotomy (n = 123) versus arthrocentesis (n = 109), and no clinical differences were observed, except for higher rate of hip SA in the former (50% vs. 9%; P < 0.001). Children with arthrocentesis had less sequelae [6.6% vs. 1%; P = 0.03, odds ratio = 0.58 (95% confidence interval: 0.45-0.76)], but not in the multivariate analysis.

This is the largest pediatric cohort of OAI in Spain. S. aureus was the most common isolate, although K. kingae was recovered in a high proportion of cases. Conservative management was applied in half of the patients. There was a low rate of sequelae, even with nonsurgical approaches.

Preservation and development of life depend on the adequate segregation of sister chromatids during mitosis and meiosis. This process is ensured by the cohesin multi-subunit complex. Mutations in this complex have been associated with an increasing number of diseases, termed cohesinopathies. The best characterized cohesinopathy is Cornelia de Lange syndrome (CdLS), in which intellectual and growth retardations are the main phenotypic manifestations. Despite some overlap, the clinical manifestations of cohesinopathies vary considerably. Novel roles of the cohesin complex have emerged during the past decades, suggesting that important cell cycle regulators exert important biological effects through non-cohesion-related functions and broadening the potential pathomechanisms involved in cohesinopathies. This review focuses on non-cohesion-related functions of the cohesin complex, gene dosage effect, epigenetic regulation and TGF-β in cohesinopathy context, especially in comparison to Chronic Atrial and Intestinal Dysrhythmia (CAID) syndrome, a very distinct cohesinopathy caused by a homozygous Shugoshin-1 (SGO1) mutation (K23E) and characterized by pacemaker failure in both heart (sick sinus syndrome followed by atrial flutter) and gut (chronic intestinal pseudo-obstruction) with no intellectual or growth delay. We discuss the possible impact of SGO1 alterations in human pathologies and the potential impact of the SGO1 K23E mutation in the sinus node and gut development and functions. We suggest that the human phenotypes observed in CdLS, CAID syndrome and other cohesinopathies can inform future studies into the less well-known non-cohesion-related functions of cohesin complex genes. Abbreviations: AD: Alzheimer Disease; AFF4: AF4/FMR2 Family Member 4; ANKRD11: Ankyrin Repeat Domain 11; APC: Anaphase Promoter Complex; ASD: Atrial Septal Defect; ATRX: ATRX Chromatin Remodeler; ATRX: Alpha Thalassemia X-linked intellectual disability syndrome; BIRC5: Baculoviral IAP Repeat Containing 5; BMP: Bone Morphogenetic Protein; BRD4: Bromodomain Containing 4; BUB1: BUB1 Mitotic Checkpoint Serine/Threonine Kinase; CAID: Chronic Atrial and Intestinal Dysrhythmia; CDK1: Cyclin Dependent Kinase 1; CdLS: Cornelia de Lange Syndrome; CHD: Congenital Heart Disease; CHOPS: Cognitive impairment, coarse facies, Heart defects, Obesity, Pulmonary involvement, Short stature, and skeletal dysplasia; CIPO: Chronic Intestinal Pseudo-Obstruction; c-kit: KIT Proto-Oncogene Receptor Tyrosine Kinase; CoATs: Cohesin Acetyltransferases; CTCF: CCCTC-Binding Factor; DDX11: DEAD/H-Box Helicase 11; ERG: Transcriptional Regulator ERG; ESCO2: Establishment of Sister Chromatid Cohesion N-Acetyltransferase 2; GJC1: Gap Junction Protein Gamma 1; H2A: Histone H2A; H3K4: Histone H3 Lysine 4; H3K9: Histone H3 Lysine 9; HCN4: Hyperpolarization Activated Cyclic Nucleotide Gated Potassium and Sodium Channel 4;p HDAC8: Histone deacetylases 8; HP1: Heterochromatin Protein 1; ICC: Interstitial Cells of Cajal; ICC-MP: Myenteric Plexus Interstitial cells of Cajal; ICC-DMP: Deep Muscular Plexus Interstitial cells of Cajal; I_f: Pacemaker Funny Current; IP3: Inositol trisphosphate; JNK: C-Jun N-Terminal Kinase; LDS: Loeys-Dietz Syndrome; LOAD: Late-Onset Alzheimer Disease; MAPK: Mitogen-Activated Protein Kinase; MAU: MAU Sister Chromatid Cohesion Factor; MFS: Marfan Syndrome; NIPBL: NIPBL, Cohesin Loading Factor; OCT4: Octamer-Binding Protein 4; P38: P38 MAP Kinase; PDA: Patent Ductus Arteriosus; PDS5: PDS5 Cohesin Associated Factor; P-H3: Phospho Histone H3; PLK1: Polo Like Kinase 1; POPDC1: Popeye Domain Containing 1; POPDC2: Popeye Domain Containing 2; PP2A: Protein Phosphatase 2; RAD21: RAD21 Cohesin Complex Component; RBS: Roberts Syndrome; REC8: REC8 Meiotic Recombination Protein; RNAP2: RNA polymerase II; SAN: Sinoatrial node; SCN5A: Sodium Voltage-Gated Channel Alpha Subunit 5; SEC: Super Elongation Complex; SGO1: Shogoshin-1; SMAD: SMAD Family Member; SMC1A: Structural Maintenance of Chromosomes 1A; SMC3: Structural Maintenance of Chromosomes 3; SNV: Single Nucleotide Variant; SOX2: SRY-Box 2; SOX17: SRY-Box 17; SSS: Sick Sinus Syndrome; STAG2: Cohesin Subunit SA-2; TADs: Topology Associated Domains; TBX: T-box transcription factors; TGF-β: Transforming Growth Factor β; TGFBR: Transforming Growth Factor β receptor; TOF: Tetralogy of Fallot; TREK1: TREK-1 K(+) Channel Subunit; VSD: Ventricular Septal Defect; WABS: Warsaw Breakage Syndrome; WAPL: WAPL Cohesin Release Factor.

The objective of this study was to examine the expression of Toll-like receptor (TLR) 3 at the maternal-fetal interface and determine whether exposure to TLR3 agonist would induce an innate immune response and trigger pregnancy loss. To address this, abortion-prone male DBA/2J mated-CBA/J female mice were given polyinosinic-polycytidylic acid (poly I:C; 10 microg/g body weight, i.p.) or PBS at gestation day (gd) 6.5. All implantation sites appeared viable at gd 7.5 when endometrium was dissected for immunohistological examination. It was noted that poly I:C treatment increased fetal losses to 40.2+/-1.7% at midgestation stage compared with control animals (11.0+/-3.0%). It was observed also that the ratio of vessel to lumen area significantly increased at gd 10.5 and gd 12.5 after poly I:C treatment, indicating that the spiral artery (SA) modification was impaired. Meanwhile, 24h after poly I:C injection, expression of TLR3 was markedly elevated within decidua basalis (DB), and endometrial TNF-alpha increased 2.7-fold but IFN-gamma remained unchanged in homogenized endometrium. These results suggest that enhanced TNF-alpha expression in endometrial stroma may play a critical role in inflammatory factor production and impairment of uterine spiral artery remodeling in the pregnancy failure of CBA x DBA/2 mating.

BACKGROUND

Autophagy enables cells to digest endogenous/exogenous waste products, thus potentially prolonging the cellular lifespan. Early endothelial progenitor cells (eEPCs) protect mice from ischemic acute kidney injury (AKI). The mid-term prognosis in AKI critically depends on vascular rarefication and interstitial fibrosis with the latter partly being induced by mesenchymal transdifferentiation of endothelial cells (EndoMT). This study aimed to determine the impact of eEPC preconditioning with different autophagy inducing agents [suberoylanilide hydroxamic acid (SAHA)/temsirolimus] in ischemic AKI.

METHODS

Male C57/Bl6 N mice were subjected to bilateral renal ischemia (40 min). Animals were injected with either untreated, or SAHA- or temsirolimus-pretreated syngeneic murine eEPCs at the time of reperfusion. Mice were analyzed 48 h and 4 weeks later. In addition, cultured eEPCs were treated with transforming growth factor (TGF)-β ± SAHA, autophagy (perinuclear LC3-II), and stress-induced premature senescence (SIPS-senescence-associated β-galactosidase, SA-β-Gal), and were evaluated 96 h later.

RESULTS

Cultured eEPCs showed reduced perinuclear density of LC3-II + vesicles and elevated levels of SA-β-Gal after treatment with TGF-β alone, indicating impaired autophagy and aggravated SIPS. These effects were completely abrogated by SAHA. Systemic administration of either SAHA or tems pretreated eEPCs resulted in elevated intrarenal endothelial p62 at 48 h and 4 weeks, indicating stimulated endothelial autophagy. This effect was most pronounced after injection of SAHA-treated eEPCs. At 4 weeks endothelial expression of mesenchymal alpha-smooth muscle actin (αSMA) was reduced in animals receiving untreated and SAHA-pretreated cells. In addition, SAHA-treated cells reduced fibrosis at week 4. Tems in contrast aggravated EndoMT. Postischemic renal function declined after renal ischemia and remained unaffected in all experimental cell treatment groups.

CONCLUSIONS

In ischemic AKI, intrarenal endothelial autophagy may be stabilized by systemic administration of pharmacologically preconditioned eEPCs. Early EPCs can reduce postischemic EndoMT and fibrosis in the mid-term. Autophagy induction in eEPCs either increases or decreases the mesenchymal properties of intrarenal endothelial cells, depending on the substance being used. Thus, endothelial autophagy induction in ischemic AKI, mediated by eEPCs is not a renoprotective event per se.

The role of the C2 protein in the pathogenicity of tomato yellow leaf curl Sardinia virus (TYLCSV) was investigated. Here we report that Agrobacterium-mediated transient expression of TYLCSV C2 resulted in a strong hypersensitive response (HR) in Nicotiana benthamiana, N. tabacum, and Arabidopsis thaliana, with induction of plant cell death and production of H2O2. Since HR is not evident in plants infected by TYLCSV, it is expected that TYLCSV encodes a gene (or genes) that counters this response. HR was partially counteracted by co-agroinfiltration of TYLCSV V2 and Rep, leading to chlorotic reaction, with no HR development. Considering that the corresponding C2 protein of the closely related tomato yellow leaf curl virus (TYLCV) did not induce HR, alignment of the C2 proteins of TYLCSV and TYLCV were carried out and a hypervariable region of 16 amino acids was identified. Its role in the induction of HR was demonstrated using TYLCSV-TYLCV C2 chimeric genes, encoding two TYLCSV C2 variants with a complete (16 aa) or a partial (10 aa only) swap of the corresponding sequence of TYLCV C2. Furthermore, using NahG transgenic N. benthamiana lines compromised in the accumulation of salicylic acid (SA), a key regulator of HR, only a chlorotic response occurred in TYLCSV C2-infiltrated tissue, indicating that SA participates in such plant defense process. These findings demonstrate that TYLCSV C2 acts as a pathogenicity determinant and induces host defense responses controlled by the SA pathway.

Glibenclamide (GLIB) is a poorly soluble drug with formulation-dependent bioavailability. Therefore, we attempted in this study to improve GLIB dissolution rate by preparing drug solid dispersions by solvent evaporation (SE) and supercritical fluid solvent-antisolvent techniques (SCF-SAS). A D-optimal mixture design was used to investigate the effects of different ratios of HPMCE5 (50-100%), PEG6000 (0-40%), and Poloxamer407 (0-20%) on drug dissolution from different solid dispersion (SD) formulations prepared by SE. The ratios of carriers used in SCF-SAS method were HPMCE5 (fixed at 60%), PEG6000 (20-40%), and Poloxamer407 (0-20%). A constant drug: carrier weight ratio of 1:10 was used in all experiments. The SDs obtained were physically characterized and subjected to the dissolution study. The major GLIB bands in FTIR spectra were indicative of drug integrity. The reduced intensity and the fewer number of peaks observed in X-ray diffractograms (XRD) of GLIB formulations was the indicative of at least partial transformation of crystalline to amorphous GLIB. This change and/or dilution of drug in much higher amounts of carriers present caused disappearance of distinctive endothermic peaks in differential scanning calorimetry thermograms of GLIB formulations. The model generated according to the results of the D-optimal mixture design indicated that GLIB formulations comprising HPMC (50%-60%), PEG (34-40%), and poloxamer (6-10%) had enhanced dissolution performances. As compared to SE method, the SCF-SAS technique produced formulations of higher dissolution performances, likely due to the effects of solution and the supercritical CO2 (SC-CO2) on enhanced plasticization of polymers and thus increased diffusion of the drug into the polymer matrix.

The aim of this study was to assess the influence of the handgrip test (HGT) on: (1) pial artery pulsation (cc-TQ), (2) subarachnoid space (SAS) width (sas-TQ) and (3) the relationship between peripheral blood pressure (BP), heart rate (HR), cerebral blood flow velocity (CBFV), resistive index (RI), cc-TQ and sas-TQ. The study was performed on 29 healthy volunteers (11 men and 18 women) with a mean age of 29.3 ± SE 4.0. HGT was performed in the sitting position at 30% of maximal voluntary contraction. cc-TQ and sas-TQ were registered using near-infrared transillumination/backscattering sounding (NIR-T/BSS); BP and heart rate (HR) were measured using a Finapres monitor. CBFV and RI were recorded using a transcranial Doppler. A significant reduction in cc-TQ (-34.3%, P<0.0001) and sas-TQ (-12.9%, P<0.001) were observed, while mean arterial pressure and HR increased (+34.8%, P<0.0001 and +7.9%, P<0.0001, respectively). There was no significant change in CBFV (+1.0 percent) while RI increased (+12.0 percent, P<0.05). Correlation and regression analysis did not reveal any interdependencies between the investigated variables. HGT evoked a significant increase in pial artery resistance, with a simultaneous decrease in the width of the SAS. A decrease in pial artery compliance should be seen as protective mechanism against acute BP elevation, most likely mediated by sympathetic activation. NIR-T/BSS recordings allowed for non-invasive assessments of changes in pial artery compliance, and were consistent with data from the literature and physiological knowledge.

OBJECTIVE

The aim of the CEntralised Pan-South African survey on tHE Under-treatment of hypercholeSterolaemia (CEPHEUS SA) was to evaluate the current use and efficacy of lipidlowering drugs (LLDs), and to identify possible patient and physician characteristics associated with failure, if any, to achieve low-density lipoprotein cholesterol (LDL-C) targets.

METHODS

The survey was conducted in 69 study centres in South Africa and recruited consecutive consenting patients who had been prescribed LLDs for at least three months. One visit was scheduled for data collection, including fasting plasma lipid and glucose levels. Physicians and patients completed questionnaires regarding their knowledge, awareness and perceptions of hypercholesterolaemia and the treatment thereof.

RESULTS

Of the 3 001 patients recruited, 2 996 were included in the final analyses. The mean age was 59.4 years, and 47.5% were female. Only 60.5 and 52.3% of patients on LLDs for at least three months achieved the LDL-C target recommended by the NCEP ATP III/2004 updated NCEP ATP III and the Fourth JETF/South African guidelines, respectively. Being male, older than 40 years, falling into the lower-risk categories, compliance with the medication regimen, and patient knowledge that the LDL-C goal had been reached, were associated with the highest probability of attaining LDL-C goals.

CONCLUSIONS

The results of this survey highlight the sub-optimal lipid control achieved in many South African patients taking lipid-lowering therapy.

This study investigated whether the myocardium is involved in the acute inflammatory reaction associated with bursts of unstable angina (UA). We looked for the presence of activated DR+ inflammatory cells and the expression patterns, localization, and immunostaining identification of genes for cytokines (IL-1beta, TNF-alpha, IL-6, and IFN-gamma), MCP-1, and iNOS in the left ventricle biopsies from 2-vessel disease anginal patients, 24 with UA and 12 with stable angina (SA), who underwent coronary bypass surgery. Biopsy specimens from 6 patients with mitral stenosis who underwent valve replacement were examined as control hearts (CHs). Plasma levels of IL-2 soluble receptor (sIL-2R) were measured as a marker of systemic immune reaction. In CHs, DR+ cells were undetectable, and cytokine and iNOS mRNA expression were negligible. UA patients had higher sIL-2R levels than SA patients (P<0.01), and their biopsy specimens showed both numerous DR+ cells identified as lymphocytes, macrophages, endothelial cells, and elevated expression levels of cytokine and iNOS genes (from 2.4- to 6.1-fold vs SA; P<0.01). Cytokine and iNOS genes and proteins were localized in endothelial cells without involvement of myocytes. IL-1beta and MCP-1 mRNAs were nearly undetectable. No significant differences were found in the number of DR+ cells, levels of cytokine, and iNOS genes between potentially ischemic and nonischemic left ventricle areas. In SA specimens, DR+ cells were very rare and only mRNAs for TNF-alpha and iNOS genes were overexpressed versus CHs. These results indicated that an acute immunomediated inflammatory reaction, essentially involving coronary microvessels, is demonstrable in UA patients.

Electroretinogramms (ERG) of isolated crayfish retinas in salines differing in their Caation were recorded to monitor changes in the ERG induced by changes in the extracellular Caation. Laser microprobe mass spectroscopy and electron microscopy of (a) shock-frozen and (b) chemically fixed retinas were used to analyse the distribution of Ca in the photoreceptor cells. For quantitative analysis a new standardisation procedure using vacuum deposition onto the specimen of thin films as an internal standard was developed. For the first time stable isotopes were used in microbeam analysis allowing direct measurements of Ca transport and metabolism on the cellular level. The major portion of Ca was found in the black distal shielding pigment granules (DP) within the retinular photoreceptor cells. Untreated retinas and retinas preincubated in physiological saline (with 10 mmol/1 Caained up to 100 mmol/1 Ca in the DP, while in DP-free places within the same cell Ca was as low as < 40 mumol/1. If the Ca-concentration of the saline was increased (decreased), a rise (fall) of Ca in the DP was observed. Careful Ca-depletion of the DP under ERG control allowed removal of estimated 60--70% of the 49Ca originally present and refilling with 44Ca. The maximum amplitude of the ERG-response decreased under these conditions to 50% in low Ca saline, but could be reestablished to some 70% in physiological saline containing 44Ca. We conclude, that in the living cell the DP acts sa a Ca store possibly regulating the intracellular and/or extracellular Ca level.

BACKGROUND

Concerns have been raised about childhood obesity and its long-term impact on the health of children. The objective of this study was to investigate rural-urban differences in body composition, energy intake, physical activity and screen time in New Zealand children.

METHODS

This study reports on data collected in a large national cross-sectional population survey of 5-15 year-old New Zealanders (the 2002 National Children's Nutrition Survey, CNS02). Schools were randomly selected to participate, as were pupils from the selected schools. Measurements of body composition were taken at school. Energy intake, physical activity and screen time information were taken from interviews and questionnaires undertaken by the child and parent/guardian. Means and standard deviations were calculated in the Statistical Analysis System (SAS Inst; Cary, NC, USA). Differences between groups were analysed using Proc Mixed after adjusting for socio-economic status and ethnicity. Data indicating differences between groups were presented as least square means +/- 95% confidence limits (CL); unless otherwise stated, and the alpha was set at p< 0.05.

RESULTS

Rural children had a significantly lower BMI, smaller waist circumferences and thinner skinfold measurements than urban children. The differences in skinfold thicknesses remained after controlling for ethnicity and socioeconomic status. Furthermore, urban boys were 1.3 times more likely to be overweight or obese than rural boys (95% confidence limits 1.1-1.6, p <0.01) and urban girls were 1.4 times more likely to be overweight or obese than rural girls (95% CL 1.2-1.7, p <0.01).There was no significant difference in the energy intake per day of rural and urban children. Similarly, there was no significant difference in the frequency of bouts of physical activity undertaken by rural and urban children.

CONCLUSIONS

Differences were found in body composition with rural children being leaner than urban children. This finding is different from other Western countries and may be due to differences in the physical and social environment in New Zealand. More research is required to understand these potential environmental rural-urban differences.