To confirm the validity of our findings, we repeated the synthesi

To confirm the validity of our findings, we repeated the synthesis of SIPPs using the fatty amine, TDA, in a ABT-263 order 30-min reflux reaction. We fully characterized both structural and magnetic properties of the second batch of TDA-SIPPs and compared the results to those of the initial batch. Table 3 shows the comparison of the two different preparations of TDA-SIPPs. Reproducibility is seen in the size JPH203 and shape of the TDA-SIPPs. Likewise, fairly good reproducibility is also seen for the other structural characteristics such as volume, surface area, concentration, and iron/platinum stoichiometry. Table 4 compares the magnetic characterizations of the

two separate TDA-SIPP preparations. Again, the reproducibility is fairly good, and the particles had similar blocking temperatures and mass magnetizations. The average mass magnetization of the TDA-SIPPs was 108.98 A m2/kg iron ± 20.38 A m2/kg iron. This value of mass magnetization was still higher than that measured

for the other SIPPs made with all of the other fatty amines examined in this study (DDA, HDA, and ODA). Table 3 Comparison of SIPPs made with tetradecylamine and a 30-min reflux Value Description Units TDA-SIPP no. 1 TDA-SIPP no. 2 d Diameter nm 7.34 ± 1.22 7.86 ± 0.76 CV Coefficient of variation % 16.6 9.6 V p Particle volume cm3 2.07 × 10−19 2.55 × 10−19 S Surface area cm2 1.69 × 10−12 1.94 × 10−12 C p Suspension concentration mg/mL 4.29 ± 0.47 5.97 ± 0.14

C Fe Iron concentration mg/mL 0.214 ± 0.00007 0.729 ± 0.004 BIRB 796 molecular weight C Pt Platinum concentration mg/mL 0.583 ± 0.0003 2.503 ± 0.005 N a Fe Iron atoms in 1.0 mL – 2.31 × 1018 7.87 × 1018 N SIPP Nanoparticles per mL SIPP/mL 5.90 × 1014 1.83 × 1015 A Fe Atomic percent Fe at.% 56.2 50.4 A Pt Atomic percent Pt at.% 43.8 49.6 Fe/Pt Fe/Pt stoichiometry – 1.28 unless 1.02 M P FePt Mass per particle g 2.9 × 10−18 3.56 × 10−18 N a FePt Total Fe + Pt atoms per particle – 6,964 8,551 N P Fe Iron atoms per particle – 3913.8 4309.9 N P Pt Platinum atoms per particle – 3050.3 4241.5 Table 4 Average magnetic properties of TDA-SIPPs ( n  = 2) Value Description Units TDA-SIPP no. 1 TDA-SIPP no. 2 Mean T b Blocking temperature K 100 150 125 ± 35.3 M sat Saturation magnetization A m2/kg iron 123.39 94.57 108.98 ± 20.38 K Effective anisotropy J/m3 1.7 × 105 2.0 × 105 1.8 × 105 ± 2.6 × 104 Conclusions Iron-platinum particles were successfully synthesized using four different fatty amines, from 12 to 18 carbons in length. Although some iron oxide contamination was seen, this decreased with increasing reflux time and decreasing chain length. Additionally, increasing the amount of time that the particles were allowed to reflux also increased the diameter of the particles, but decreased the iron concentration.

On average, these

On average, these subjects possessed 23 teeth. The control group consisted of 20 healthy volunteers aged 46.5 ± 6 years (eight women and 12 men), matched by body mass index (BMI), without signs of pathological tooth wear. They were asked

to participate voluntary in the study as they presented at the department to make minor prosthetic procedures relating only to a single tooth (e.g., crown or inlay). On average, the reference subjects possessed CP-690550 research buy 27 teeth. All study participants demonstrated good general somatic condition, their own teeth were free of clinical signs of dental caries or periodontal disease. Tooth Wear Index (TWI) was used to categorize the participants. The inclusion criteria for the studied patients were: the presence of widespread advanced tooth wear with multiple sites

of exposed occlusal dentin (TWI on occlusal/incisal surface ≥ 2) and a considerable decrease in the occlusal vertical dimension (more than 4 mm measured in the anterior region), no significant periodontal bone loss or decay on their own teeth, no prior prosthetic rehabilitation attempting to treat the lost vertical dimension. Furthermore, the inclusion criteria for the whole group encompassed: absence of chronic metabolic, endocrine, renal, or gastrointestinal conditions, or prolonged medication known to selleckchem affect bone metabolism, selleck inhibitor oral microflora, or the salivary flow rate. No history of clinically significant fractures was reported in either of the groups. Patients who underwent prosthetic rehabilitation prior to recruitment were excluded. The participants were required to be available to be recalled multiple times during the duration of the study.

Informed consent was obtained from Pregnenolone each participant prior to confirmation of their eligibility for the study. The protocol was approved by the Ethical Committee of the Medical University of Bialystok, Poland (approval # R-I-003/6/2006). Methods The dietary intakes of nutrients, including elements and vitamin D, were assessed with the validated 7-days food-frequency questionnaire based on the software DIETA 3, and the obtained data were compared to the national recommended daily intakes (RDIs). Both direct standardized interview-based questionnaire and medical records were utilized to accomplish medical history. Anthropometric measurements (body weight and height) were performed using electronic scale (Seca, Germany) and Harpenden stadiometer, whereas BMI was calculated using standard formula. Dental examination and clinical procedures All of the participants were clinically examined to evaluate tooth wear. Tooth wear was assessed according to the protocol of Smith and Knight [42]. The TWI was selected as an assessment measure because this method allowed to visually evaluate the level of wear.

The results of both methods were not significantly different and

The results of both methods were not significantly different and both methods were judged suitable for the purpose of analyzing C188-9 ic50 saliva samples for acetaldehyde. While the GC method is more precise, sensitive and selective, we used the enzymatic assay for

approximately half of the samples to be analyzed, because of its lower costs and faster analysis times. Statistics All data were evaluated using Unscrambler X version 10.0.1 (Camo Software AS, Oslo, Norway) and Origin V.7.5 (Originlab, Northampton, USA). Data are summarized as means and standard deviations between assessors for each data point. Statistical dependence between alcoholic strengths and the acetaldehyde contents of the beverages and the salivary acetaldehyde were evaluated using multiple linear PARP activity regression (MLR) and Analysis of Variance (ANOVA) for all time data points (30 sec, 2 min, 5 min, and 10 min). The regression analysis was also conducted with the area under

the curve (AUC) for the complete time period under investigation (0-10 min). Statistical significance was assumed at below the 0.05 probability level. Results selleck chemical Table 1 shows the alcoholic strengths and acetaldehyde contents of the alcoholic beverages, as well as the resulting average salivary acetaldehyde concentrations for the assessors. The assessors (up to n = 10 per beverage, see Table 1) had an average age of 27 ± 6 years and 70% were female. The highest salivary acetaldehyde concentration was found in the saliva 30 sec after using the beverages in all cases, and the average content was 353 ± 164 μM (range: 56-1074 μM). The acetaldehyde level then decreased at the 2-min sampling (156 ± 46 μM, range: 41-337 μM), the 5-min sampling (76 ± 19 μM, range 26-131 μM) and at the 10-min sampling (40 ± 18 μM, range: n.d.-94 μM). The inter-individual variation in salivary acetaldehyde content is relatively high, with an average CV of 48% between assessors. No apparent gender or age related differences

were seen, however, due to the relatively homogenous ages of the probands, the statistical Dehydratase power does not allow to make a definite conclusion on an effect of age. Similarly, no statistically significant conclusion on the effect of gender can be gathered from the data. Table 1 Alcoholic strength and acetaldehyde content of alcoholic beverages and the resulting salivary acetaldehyde concentrations         Salivary acetaldehyde [μM]a Alcoholic beverage Alcoholic strength [% vol] Acetaldehyde b [μM] Number of assessors f 0.5 min 2 min 5 min 10 min Beerc 5 210 1 98 ± 4 113 ± 13 44 ± 6 n.d.e Ciderc 5.5 2529 4 428 ± 159 202 ± 72 70 ± 41 26 ± 7 Winec 13 474 3 315 ± 288 225 ± 117 115 ± 62 39 ± 30 Calvadosd 15g 411 2 93 ± 59 51 ± 16 27 ± 10 n.d.e Sherryc 15 2583 3 291 ± 117 114 ± 77 68 ± 25 n.d.e Vodkad 16g n.d. 3 56 ± 11 59 ± 30 36 ± 27 n.d.

07) Figure 2c demonstrates that there was no difference in the o

07). Figure 2c demonstrates that there was no difference in the overall length of stay (Mann-Whitney U test, p = 0.072), duration of delay to surgery (Mann-Whitney U test, p = 0.35) and length of postoperative stay in hospital (Mann-Whitney

U test, p = 0.25). Figure 2 Comparison of time from admission to surgery (a), postoperative length of stay (b) and total length of stay (c) between the two groups. Box and whisker graphs represent median ± inter-quartile range. Discussion Our audit in a comparable cohort of patients over two different time periods, after a change in theatre prioritisation policy, did not demonstrate any significant differences in the outcome after appendicectomy. The intention of implementing this change was to effectively reduce waiting times to emergency surgery and hence length of hospital stay – but clearly the present study has failed E7080 cell line to demonstrate this effect. There could be numerous reasons for this finding. Foremost, this could be due to the small sample size, which will require CP673451 cell line a multi-centre study.

Such a study could be hampered by AZD5582 mouse non-homogeneity of the profile of emergency workload. Our hospital is one of the premier trauma units in the UK and the only site of the only Helicopter emergency medical service (HEMS) in London. Despite this, numerically at least emergency general surgery accounts for 64.2% of all the emergency surgical workload with abscesses and acute appendicitis being the two most frequent reasons for requiring theatre [11]. Of course, trauma as well as vascular operations, because of the complexity of pre-operative and operative work and multiple team involvement, take longer duration and therefore occupy a prominent part of the emergency theatre schedule. Some authors have suggested an increase in post-appendicectomy complications and longer hospital stay associated to the delay to surgery [12, 13], whilst others have failed to demonstrate this trend [14–17]; although, LY294002 of course most patients would

prefer immediate surgical procedure [18]. In our cohort only four patients had a complication; of those, three were operated within 10 hours from admission and only one after 18 hours. Our data doesn’t demonstrate significant changes in outcome after the appendicectomy, despite changes in theatre prioritisation. The median length of hospital stay was 76 hours, comparable to other publications [13, 14]. Delay to surgery is associated with an increased incidence of complications and length of hospital stay after appendicectomy [12, 13, 19]. Analyzing a large series of 1081 patients, Ditillo et al[12] from the Yale University, USA demonstrated that in adult patients with acute appendicitis, the risk of developing advanced pathology and postoperative complications increases with time; particularly, those risks rise proportional to delay.

Embryos were collected and chilled every 15 minutes until approxi

Embryos were collected and chilled every 15 minutes until approximately 200 μl of packed embryos

were obtained per replicate. The eggs were stored at -80C until DNA extractions could be performed. Synchronization of larvae was accomplished by allowing several hundred females to oviposit on egg-laying dishes for one hour. The eggs were collected and seeded onto standard media. From these, third instar (3′) larvae were collected and stored at -80C until DNA extraction. DNA was extracted from all tissues and flies with the DNeasy Blood and Tissue Kit (Qiagen) using the manufacturer’s protocol with an extended, overnight Nepicastat proteinase K digestion. DNA purity and concentration was determined using a Nanodrop ND1000. Quantitative PCR for relative copy number Relative copy numbers of Wolbachia and WO phage in .D. simulans were obtained using the MiniOpticon System (Bio-Rad). The relative Wolbachia infection level was measured by comparing the copy number of the gene for Wolbachia JPH203 surface protein, wsp, to a single copy gene in the Drosophila genome, CuZn superoxide dismutase (sod). Phage copy numbers were measured by comparing the adenine methyltransferase (wMTase) (WORiB), lyzozyme (WORiA), and tail tube protein (WORiC) genes to wsp in wRi (see table 1 for

locus tags and primer sequences). Table 1 Primer sequences used in this study ORF Product Locus Tag Specificity Sequence (5′-3′) Superoxide Dsim GD12822 D. simulans F – GTCGACGAGAATCGTCACCT Dismutase (SOD)     R – GGAGTCGGTGATGTTGACCT Surface Antigen WRi 010990 Wolbachia F – ATCAGGGTTGATGTTGAAGG

Wsp (Wsp)   w Ri R – CAGTATCTGGGTTAAATGCTG Lyzozyme M1 WRi 012650 WORiA F – GACTTTATGGCAGTATACCGA (Lyz)     R – TGTTCCGTTGAATTTGTTCC DNA WRi 005640 WORiB F – CTTAAATGACCATCAACCACAG Methyltransferase (MTase)     R – GCTTCAATCAGGGAATTTGG Metalloexopeptidase Contractile Tail WRi 006970 WORiC F- GTTGATGGTAGAGGTTATGCAG Tube Protein     R – GAATATCCATACCACCAGCTC Reactions were performed in low profile 48-well white plates with flat cap strips (Bio-Rad). Ten microliter reactions included 400nM of each forward and reverse primer, 5 μl of 2× Dynamite qPCR mastermix (YH25448 in vivo Molecular Biology Service Unit – University of Alberta) which included SYBR green (Molecular Probes) and Platinum Taq (Invitrogen), and 125ng of DNA. The thermal cycling conditions were 95°C for 2 minutes, 40 cycles of 95°C, 55°C, and 72°C for 30 seconds each, and a final 2 minute 72°C extension. Fluorescent data were acquired after every 72°C extension. A 60-95°C melting curve was performed to confirm the specificity of the products. No template controls were included to account for DNA contamination. All samples were analyzed in technical and biological triplicates.

Continuous, uniform, and crack/void-free CoFe2O4/polymer films wi

Continuous, uniform, and crack/void-free CoFe2O4/polymer films with thicknesses in the range 200 nm to 1.6 μm were systematically this website prepared by multiple spin/cast coating followed by thermal treatment to dry the film. Figure  3 shows SEM images with a CFO weight fraction of 25%

where the white dots are the CFO nanoparticles and the dark background is the P(VDF-HFP) copolymer. The top surface view of the microstructure of the nanocomposite film demonstrates that monodisperse, ultrafine cobalt ferrite HCS assay nanoparticles are well embedded in the polymer matrix, forming typical 0–3, particulate type nanocomposites. Loose agglomeration occurs locally due to the magnetic interaction among the nanopowders. Defects, pores, or phase separation unfavorable for device fabrication was not observed. The cross-sectional image (Figure  3b) confirms the thickness of the free standing film of approximately 1.5 μm. The observation of intimate physical contact between the CFO and P(VDF-HFP) phase components is a good starting point for attempting to generate mechanical, magnetic, or electrical coupling between them. Figure 3 SEM images of CoFe 2 O 4 / P ( VDF-HFP ) thin-films deposited on Si substrate. With cobalt ferrite

fraction of 25 wt.% and film thickness of 1.5 μm. (a) Top surface view; (b) cross-sectional view. The effective permittivity (ϵ eff) and loss tangent (tan δ) of the ferrites/polymer thin films (thickness of approximately 1 μm) were measured over the frequency range from 100 Hz to 1 MHz (Figure  4). Both the effective permittivity and loss tangent of the nanostructured films selleck products show a systemic increase as a function

of the loading of CFO nanocrystals. The dielectric constant of the pure P(VDF-HFP) film is measured to be 8 at 100 Hz (Figure  4a), consistent with the reported data [24, 25], and increases to 44 in the case of the 30 wt.% CFO samples due to the inclusion of the higher dielectric constant magnetic component (k(CoFe2O4) ≈ 400) [26]. The polarization in ferrites originates from the electronic exchange Fe2+ ⇔ Fe3+ and hole transfer between Co2+ ⇔ Co3+ in the spinel phase, which cannot follow the alternating external field beyond a certain frequency [27]. When Selleckchem Gemcitabine the space charge carriers fail to keep up with the field and lag behind the alternation of its direction, the composites’ permittivity and loss tangent decrease monotonically with frequency. Once the frequency is over 10 kHz, the relaxation mechanism associated with the P(VDF-HFP) phase dominates the overall dielectric behavior [20]. The decrease in loss (Figure  4b) with frequency at low frequencies (<1 kHz) is attributed to the ionic DC conduction contribution from the P(VDF-HFP) copolymer phase, which yields interfacial or spatial charge polarization [28]. The increase in loss at high frequencies (>10 kHz) results from the β relaxation associated with the glass transition of the copolymer.

One of our sequences affiliated with Crenarchaea cluster 1 1b, wh

One of our sequences affiliated with Crenarchaea cluster 1.1b, which includes several putative AOA [54–56]. However, it has recently been shown that not all amoA-carrying Thaumarchaeota are ammonia-oxidizing autotrophs [57]. The presence

of AOA at the Rya WWTP can therefore not be confirmed, and as has been suggested for other WWTPs [14, 16], AOA are most likely of minor or no importance for ammonia-oxidation at the Rya WWTP. One clone affiliated with Crenarchaea cluster 1.3. There are no cultured representatives of cluster 1.3, but spatial co-localization [58] and a relation between the abundance of cluster 1.3 and Methanosaeta-like species has been reported [42]. In other aggregate structures, such as anaerobic sludge Fedratinib price selleck chemicals granules, Methanosaeta are important for structure and stability and they form dense aggregates which act as nuclei for granule formation [20]. In the activated sludge the Methanosaeta did not appear to have this function as they were mostly detected as small colonies or single cells (Figure  11) and there was no apparent difference

in structure between flocs with high and low numbers of Methanosaeta. The lowest relative abundances of the Methanosaeta-like TRFs were observed in RSL3 order January and February 2004 (Figures  7 and 8). In October 2003 the two main Methanosaeta TRFs also decreased in relative abundance but it cannot be ruled out that the TRFs that appeared in those samples were also Methanosaeta (Table 4). The lowest water temperatures of the period were recorded during January and February 2004, which could have

reduced the survival or proliferation of Methanosaeta-like species and allowed other Archaea to increase. In anaerobic sludge, a decrease in Methanosaeta abundance has mafosfamide been linked to granule disintegration [18, 19]. Although the flocs had high shear sensitivity and a more open structure in January and February 2004 when the Methanosaeta TRFs decreased and although there was a significant negative correlation between Methanosaeta TRFs and effluent non-settleable solids (Table 6) it cannot be concluded that the Archaea are important for the floc structure. The increased shear sensitivity and changed floc structure in January and February 2004 could be due to the reduced general microbial activity, which has been shown to decrease floc stability [5]. Furthermore, increased shear sensitivity and changed floc structure was also observed from June to August 2004, after the primary settlers were bypassed, but during this period the relative abundance of the Methanosaeta TRFs was 100%. Thus, if the composition of the Archaea community has any effect on floc structure or stability it is certainly only one of many other factors. Conclusions By sequencing and T-RFLP analysis of 16S rRNA genes and FISH we showed that Archaea were present in the activated sludge of a full-scale WWTP.

Lymph node metastasis (pN+,

Lymph node metastasis (pN+, Selleck LY2874455 p < 0.0001, Hazard Ratio (HR) = 12.1940, 95% CI = 5.9509 - 24.9867), pT-category (pT3/4, p < 0.0001, HR = 3.8447, 95% CI = 1.5309 - 9.6553) and grading (G3/4, p < 0.0001, HR = 4.0652, 95% CI = 1.7123 - 9.6514) were shown to be unfavorable factors in univariate analysis in the whole RAD001 purchase population of all EACs (n = 60). Survival in subgroup with high LgR5 expression in BE (n = 41, p = 0.0278, HR = 3.5145, 95% CI = 1.5050 – 8.2073, Figure 4a), adjacent EACs (n = 41, p = 0.039, HR = 2.8408, 95% CI = 1.2496 – 6.4582) and all EACs (n = 60, p = 0.0325, HR = 2.4175, 95% CI = 1.1719 – 4.9872, Figure 4b) was significantly

poorer in comparison to the subgroup of patients with low expression of LgR5

(Table 1 and 2). Data suggest that LgR5 expression in BE and adjacent EACs is associated with clinical pathological STA-9090 research buy features which may predict worse clinical outcome of related (adjacent) adenocarcinomas. Multivariate analysis using the Cox Proportional Hazards Model demonstrate lymph node metastasis and grading but not LgR5 expression as independent prognostic factors in all (n = 60) EACs (LN positive: Exp (b) 9.1861; 95% CI of Exp (b) 2.0665 – 40.8346; p = 0.003746. Grading G3/4: Exp (b) 2.2593; 95% CI of Exp (b) 1.0171 – 5.0186; p = 0.4643). Figure 4 Kaplan-Meier survival curves. Overall survival curves calculated by Kaplan-Meier Farnesyltransferase method in Barrett-associated EACs (Figure 4a) and the whole population of all EACs (Figure 4b), respectively. Survival of patients with EAC was better when BE showed low LgR5 expression compared to high LgR5 expression. This was shown for BE in association with EAC (p = 0.0278) (a) and the whole population of EACs (b), respectively (p = 0.0325). The times of the censored data are indicated

by short vertical lines. Discussion Similar to other solid tumor entities [8], a stem cell hypothesis has been proposed for Barrett’s esophagus (BE) and its association with EAC [13]. However, this hypothesis has not undergone thorough investigation so far. An intestinal stem cell marker, LgR5 has been proposed [13], but have also not been thoroughly addressed in histogenetic studies. Our results of LgR5 expression in EAC with and without BE, as well as the adjacent Barrett mucosa suggest that LgR5 might be a promising marker to further address the stem cell hypothesis. In esophageal SCC – as expected no LgR5 expression was found, which is due to the fact that ESCC is not derived from an intestinal (glandular) type epithelium. Several studies have already focused on the effects of different LgR5 expression in the context of tumor development and progression.

Recently, we have shown that the extent of systemic inflammation

Recently, we have shown that the extent of systemic inflammation of innate immune cells can be visualized by measuring the expression of activation markers on blood PMNs [9]. The most sensitive marker turned out to be the responsiveness of active FcγRII (CD32) on PMN’s for the innate immune stimulus fMLP [9, 10]. The most commonly used marker is MAC-1 (CD11b), which peaks between 6 and 18 hours after insult (i.e. trauma or surgery)[11]. In contrast to PMN’s, changes in activation of the systemic 3-deazaneplanocin A monocyte compartment can be determined by analyzing the percentage of circulating Bafilomycin A1 HLA-DR positive monocytes [7]. Blood samples

were taken at two distinct time points: one hour prior to IMN and 18 hours after the intramedullary nail was introduced. To investigate the influence of IMN, patients were stratified by isolated femur fracture and femur fractures

in multitrauma. Patients were compared with healthy, age and gender matched controls as described previously (see Table 1)[9]. Table 1 Patient demographics.   Median (+ range) Number of patients (n) 38 Male/Female (n) 22/16 Combretastatin A4 datasheet Age (years) 30 (16-80) Injury Severity Score 13 (9-43) – Femur fracture (n = 23) 10 (9-19) – Multitrauma (n = 15) 29 (16-43) APACHE II Score 5 (0-24) Time on ICU (days) 0 (0-60) Time on ventilation (days) 0 (0-55) Packed red blood cells before first blood sample (units) 0 (0-22) Fresh frozen plasma before first blood sample (units) 0 (0-20) Trauma mechanism (n)      - MVA 29    - Fall of height 8    - Direct impact 1 Complications (n)      - No SIRS symptoms 14    - SIRS 17    - ALI/ARDS 7 Materials For analysis of PMN receptor expression

by flowcytometry the following monoclonal antibodies were commercially purchased: FITC-labeled IgG1 negative control (clone DD7, Chemicon, Hampshire, United Kingdom), RPE-labeled IgG2a negative control (clone MRC OX-34, 4-Aminobutyrate aminotransferase Serotec, Dusseldorf, Germany) and RPE-labelled CD11b (clone 2LPM19c, DAKO, Glostrup, Denmark). An antibody, which recognizes an active FcyRII/CD32 (designated FcyRII*), is manufactured at the Department of Pulmonary Science at the University Medical Center Utrecht (MoPhab A27, UMCU, Utrecht, The Netherlands)[12, 13]. For analysis of monocyte HLA-DR expression by flowcytometry the following monoclonal antibody was commercially purchased: FITC-labeled HLA-DR (YE2/36-HLK, Serotec, Dusseldorf, Germany). Pmn and monocyte receptor expression The inflammatory status of a patient can be assessed by analyzing the expression of active FcyRII (FcyRII*) on PMNs in the peripheral blood [9]. A low expression of fMLP induced FcyRII* correlates with increased inflammation. This approach has been validated in a previous study [9]. The expression of fMLP induced FcyRII* was compared with a more common activation marker MAC-1 (CD11b)[14].

The similar reaction of diquinodithiin 1 with hydrochlorides of 1

The similar reaction of diquinodithiin 1 with hydrochlorides of 1-naphthylamine, 2-naphthylamine, and 6-aminoquinoline gave pentacyclic 7H-quinonaphthothiazine 4, 14H-quinonaphthothiazine 5, and 7H-diquinothiazine 6. The reaction of isomeric diquinodithiin 7 with acetamide and p-fluoroaniline hydrochloride gave linearly condensed pentacyclic 6H-diquinothiazines 9a and 6-(p-fluorophenyl)diquinothiazine 9b (Scheme 2). Analogous reaction of another isomeric diquinodithiin 10 with p-fluoroaniline hydrochloride led to angularly condensed diquinothiazine 12c. Better yields of the fluoroaniline products 9b and 12c were achieved when x,x’-dichloro-3,3′-diquinolinyl sulfides 8 and 11 (x = 2 and

4) were used. Sulfide 11 reacted also with ammonia or methylamine in hot phenol to give diquinothiazines

12a, b. Scheme 1 Reactans: a C6H5NH2·HCl CX-6258 concentration (p-ClC6H4NH2·HCl, p-CH3OC6H4NH2·HCl), 200–205 °C, 4 h; b p-CH3OC6H4NH2, MEDG, reflux, 3 h; c 1-naphthylamine·HCl, 200–205 °C, 4 h; d 2-naphthylamine·HCl, 200–205 °C, 4 h; e 6-aminoquinoline·HCl, 200–205 °C, 4 h Scheme 2 Reactans: a CH3CONH2, K2CO3, 180 °C, 0.5 h; b pF-C6H4NH2·HCl), 200–205 °C, 3 h; c p-FC6H4NH2, MEDG, reflux, 3 h; d NH3 (CH3NH2), phenol, 180 °C, 1 h The described syntheses were monitored by TLC analysis. All chromatograms of new compounds showed characteristic for azaphenothiazines (Jeleń et al., 2011) color changing during irradiation with UV light from blue to yellow (4, Adenosine triphosphate 9b), from yellow to green (5, 6), from orange to yellow (12c), and from yellow to orange (7c). Structure It is well Nutlin-3a mw known that the synthesis of phenothiazines can proceed via the Smiles rearrangement of the S–N type of the appropriate sulfide (Pluta et al., 2009). The identification of the product structures was based on the spectroscopic 1H NMR and MS analysis. In the case of the reactions of sulfides 7 and 11, the products 9 and 12 possessed the C2v symmetry (the left part was a mirror image of the right one) what excluded the stage of rearrangement. The reactions of diquinodithiin 1 and disulfide 2 with anilines proceeded similarly

without the stage of rearrangement to give tetracyclic quinobenzothiazines 3a–c (Jeleń and Pluta, 2009). The reaction with 1-naphthylamine gave pentacyclic quinonaphthothiazine 4. On the contrary, the reactions with 2-naphthylamine and 6-aminoquinoline were more complex as there were two possibilities of the thiazine ring formation. The 1H NMR analysis of the reaction products pointed at compounds 5 and 6 excluding compounds 13 and 14, as evidenced from coupling Wortmannin constants; the H-5 and H-6 protons in compounds 5 and 6 showed a coupling constant J ortho, whereas analogous protons in compounds 13 and 14 (H-7/H-12 and H-5/H-14, respectively) would have shown a coupling constant J para, which is very small (i.e., J 1,4 = 0.6-0.