Circulating tumor cells (CTCs) are promising biomarkers for diagnosis and therapy in systemic cancer. However, their infrequent and unreliable detection, especially in nonmetastatic cancer, currently ...impedes the clinical use of CTCs. Because leukapheresis (LA) targets peripheral blood mononuclear cells, which have a similar density to CTCs, and usually involves processing the whole circulating blood, we tested whether LA could substantially increase CTC detection in operable cancer patients. Therefore, we screened LA products generated from up to 25 L of blood per patient in two independent studies, and found that CTCs can be detected in more than 90% of nonmetastatic breast cancer patients. Interestingly, complete white blood cell sampling enabled determining an upper level for total CTC numbers of about 100,000 cells (median, 7,500 CTCs) per patient and identified a correlation of CTC numbers with anatomic disease spread. We further show that diagnostic leukapheresis can be easily combined with the US Food and Drug Administration-approved CellSearch system for standardized enumeration of CTCs. Direct comparison with 7.5 mL of blood revealed a significantly higher CTC frequency in matched LA samples. Finally, genomic single-cell profiling disclosed highly aberrant CTCs as therapy-escaping variants in breast cancer. In conclusion, LA is a clinically safe method that enabled a reliable detection of CTCs at high frequency even in nonmetastatic cancer patients, and might facilitate the routine clinical use of CTCs as in the sense of a liquid biopsy. Combined with technologies for single-cell molecular genetics or cell biology, it may significantly improve prediction of therapy response and monitoring of early systemic cancer.
Tris(bis(phenylmethyl)carbamodithioato-S,S'), commonly referred to as tris(N,N-dibenzyldithiocarbamato)indium(III), In(S2CNBz2)3, was synthesized and characterized by single crystal X-ray ...crystallography. The compound crystallizes in the triclinic space group P1 with two molecules per unit cell. The material was further characterized using a novel analytical system employing the combined powers of thermogravimetric analysis, gas chromatography/mass spectrometry, and Fourier transform infrared (FT-IR) spectroscopy to investigate its potential use as a precursor for the chemical vapor deposition (CVD) of thin film materials for photovoltaic applications. Upon heating, the material thermally decomposes to release CS2 and benzyl moieties in to the gas phase, resulting in bulk In2S3. Preliminary spray CVD experiments indicate that In(S2CNBz2)3 decomposed on a Cu substrate reacts to produce stoichiometric CuInS2 films.
Fetal hemoglobin (HbF) production induced by hydroxyurea is the mainstay of treatment for sickle cell anemia (SCA). Increased HbF production correlates with a higher number of HbF-containing red ...blood cells (RBCs) called F-cells. Successful treatment with hydroxyurea is associated with an increased number of F-cells, less hemolysis, improvement of anemia, and decreased frequency of vaso-occlusive crises in SCA patients. Comparison of in vitro sickling among blood specimens from sickle mouse models and from patients with different HbF levels has provided compelling evidence that increasing the percentage of circulating F-cells is associated with improvement of hemolytic biomarkers. While it has been demonstrated that higher HbF content prolongs sickle RBC survival, there is only indirect evidence of the response to hypoxia of F-cells compared to non-F-cells. We investigated the influence of HbF content on sickling through our recently developed Sickle Imaging Flow Cytometry Assay (SIFCA). SIFCA allows simultaneous analysis of both expression of intracellular proteins and morphological features of each cell in a robust, reproducible, operator-independent sickling assay. Peripheral venous blood samples were collected upon written consent from adult SCA patients with a wide range of HbF percentages (HbF range 2.0-26.9%) (n=15, nine on hydroxyurea treatment). RBC pellets were used to prepare 1% suspensions that were subjected to deoxygenation for 2 hours at 2% oxygen. RBCs were then labeled for HbF using a standard protocol for F-cell quantitation and a minimum of 20,000 cells were analyzed by imaging flow cytometry (ImageStreamX Mk II, Amnis Corporation), allowing the correlation between shape change and intensity of HbF expression for each RBC. We confirmed previous observations using conventional flow cytometry that F-cell count percentages significantly correlate with mean HbF determined by HPLC (r2P=0.9700, 95% CI 0.9098-0.9902, P<0.0001). F-cell count by SIFCA correlated highly with conventional F-cell flow cytometry by an independent CLIA-certified facility (r2P =0.9976, 95% CI 0.9861-0.9996, P<0.0001). SIFCA morphological analysis showed that the percentage of non-F-cells sickling upon deoxygenation was significantly higher than among F-cells (17.75% 95% CI 12.5-23.00 vs. 12.41% 95% CI 8.67-16.15, P=0.0015), a 1.498-fold difference (95% CI 1.228-1.768). Image analysis also allowed us to identify the presence of F-cells that still sickle despite their high HbF content, as well as non-F-cells that are resistant to sickling (Figure 1). Transmission electron microscopy of F-cells enriched by fluorescence activated cell sorting confirmed that sickled F-cells contained hemoglobin S polymers. In summary, we have documented for the first time at the individual RBC level that human F-cells are less prone to sickle under hypoxia ex vivo than non-F-cells. This study also illustrates the power of imaging flow cytometry to characterize predisposition to sickling in populations of red blood cells from the same patient, and would be suitable for use as a supportive biomarker assay in clinical trials investigating the efficacy of novel HbF inducers and their anti-sickling effect in a single assay. While the finding that F-cells sickle less than non-F-cells is not unexpected, it seems surprising to us that the difference in hypoxia-induced sickling between F-cells and non-F-cells is so small. This finding emphasizes the need to characterize additional RBC features that render individual cells more susceptible or resistant to sickling. Identification of factors besides HbF that modulate sickle hemoglobin polymerization may help design novel therapies for hydroxyurea-resistant SCA patients. Display omitted
No relevant conflicts of interest to declare.
Tris(bis(phenylmethyl)carbamodithioato-
S,
S′), commonly referred to as tris(
N,
N-dibenzyldithiocarbamato)indium(III), In(S
2CNBz
2)
3, was synthesized and characterized by single crystal X-ray ...crystallography. The compound crystallizes in the triclinic space group
P
1
¯
with two molecules per unit cell. The material was further characterized using a novel analytical system employing the combined powers of thermogravimetric analysis, gas chromatography/mass spectrometry, and Fourier transform infrared (FT-IR) spectroscopy to investigate its potential use as a precursor for the chemical vapor deposition (CVD) of thin film materials for photovoltaic applications. Upon heating, the material thermally decomposes to release CS
2 and benzyl moieties in to the gas phase, resulting in bulk In
2S
3. Preliminary spray CVD experiments indicate that In(S
2CNBz
2)
3 decomposed on a Cu substrate reacts to produce stoichiometric CuInS
2 films.
The synthesis and structural characterization of a novel In(III) complex is described. The reaction between InCl
3
with sodium mercapto-acetic acid (NaSCH
2
(CO)OH) in 4-methylpyridine (CH
3
(C
5
H
5
...N), (4-Mepy)) at 25°C affords ClIn(SCH
2
(CO)O)
2
2-
(4-MepyH)
2
2+
(1). X-ray diffraction studies of (1) show it to have a distorted square-pyramidal geometry with the (
-
SCH
2
(CO)CO
-
) ligands in a trans conformation. The compound crystallizes in the P1(No. 2) space group with a = 7.8624(6) Å, b = 9.950(1) Å, c = 13.793(2) Å, α = 107.60(1)°, β = 90.336(8)°, γ = 98.983(9)°, V = 1014.3(4) Å
3
, R(F°) = 0.037 and R
w
= 0.048.
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Dostopno za:
BFBNIB, DOBA, GIS, IJS, IZUM, KILJ, KISLJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
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