Caspase-dependent apoptosis is induced by Artemisia afra Jacq. ex Willd in a mitochondria-dependent manner after G2/M arrest

Artemisia afra is one of the oldest, most well known and widely used traditional medicinal plants in South Africa. It is used to treat many different medical conditions, particularly respiratory and inflammatory ailments (Liu et al., 2009). There is no reported evidence of its use for the treatment of cancer but due to its reported cytotoxicity (Fouche et al., 2008, Mativandlela et al., 2008), we investigated the mode of cell death induced by an ethanolic A. afra extract by using two cancer cell lines. IC₅₀ values of 18.21 and 31.88 μg/mL of ethanol extracts were determined against U937 and HeLa cancer cells, respectively. An IC₅₀ value of the aqueous extract was greater than 250 μg/mL. The effect of the cytotoxic ethanolic A. afra extract on U937 and HeLa cells and their progression through the cell cycle, apoptosis and mitochondrial membrane potential were investigated. Melphalan was used as a positive control. After 12 h of treatment with A. afra a delay in G2/M phase of the cell cycle was evident. Apoptosis was confirmed by using the TUNEL assay for DNA fragmentation, as well as fluorescent staining with annexin V-FITC. Apoptosis was evident with the positive control and A. afra treatment at 24 and 48 h. JC-1 staining showed a decrease in mitochondrial membrane potential at 24 h. The results obtained suggest that A. afra potentially has medicinal anticancer properties.     

Kinetic study on enzymatic esterification of tuna fish oil fatty acids with butanol

Docosahexaenoic acid (DHA) is an important polyunsatured fatty acid (PUFA) which can be purified from tuna fish oil fatty acids by selective enzymatic esterification. The present paper investigates the kinetic study for selective esterification of tuna fish oil fatty acids with butanol catalyzed by Rhizopus oryzae lipase (ROL) in biphasic solvent system. Under the most suitable reaction conditions, 76.2% esterification was achieved in 24 h. Different kinetic models for esterification given by Segel [1], Oliveira et al. [2], Gogoi et al. [3], and Kraai et al. [4] were tested for fitting the esterification data and the model given by Oliveira et al. [2] was found to be most suitable. The model given by Prazeres et al. [5] for hydrolysis was also tested for esterification and the model with second order product inhibition was found to provide better match between the predicted and experimental values than that of model by Oliveira et al. [2]. The kinetic model was fitted using MATLAB^® to determine the best kinetic parameters. The average value of kinetic constants using the model given by Prazeres et al. were estimated as K_m = 23.6 μmoles FFA/ml, K_i1 = 4.6 × 10⁻⁵ μmoles FFA/mg enzyme h, K_i2 = 0.0062 μmoles FFA/mg enzyme h and K₂ = 149.5 μmoles FFA/mg enzyme h.     

Triterpene glycosides from Astragalus icmadophilus

Six cycloartane-type triterpene glycosides were isolated from Astragalus icmadophilus along with two known cycloartane-type glycosides, five known oleanane-type triterpene glycosides and one known flavonol glycoside. The structures of the six compounds were established as 3-O-[α-L-arabinopyranosyl-(1 → 2)-O-3-acetoxy-α-L-arabinopyranosyl]-6-O-β-D-glucopyranosyl-3β,6α,16β,24(S),25-pentahydroxycycloartane, 3-O-[α-L-rhamnopyranosyl-(1 → 2)-O-α-L-arabinopyranosyl-(1 → 2)-O-β-D-xylopyranosyl]-6-O-β-D-glucopyranosyl-3β,6α,16β,24(S),25-pentahydroxy cycloartane, 3-O-[α-L-arabinopyranosyl-(1 → 2)-O-3,4-diacetoxy-α-L-arabinopyranosyl]-6-O-β-D-glucopyranosyl-3β,6α,16β,24(S),25-pentahydroxycycloartane, 3-O-[α-L-arabinopyranosyl-(1 → 2)-O-3-acetoxy-α-L-arabinopyranosyl]-6-O-β-D-glucopyranosyl-3β,6α,16β,25-tetrahydroxy-20(R),24(S)-epoxycycloartane, 3-O-[α-L-arabinopyranosyl-(1 → 2)-O-β-D-xylopyranosyl]-6-O-β-D-glucopyranosyl-3β,6α,16β,24α-tetrahydroxy-20(R),25-epoxycycloartane, 3-O-[α-L-rhamnopyranosyl-(1 → 2)-O-α-L-arabinopyranosyl-(1 → 2)-O-β-D-xylopyranosyl]-6-O-β-D-glucopyranosyl-3β,6α,16β,24α-tetrahydroxy-20(R),25-epoxycycloartane by the extensive use of 1D- and 2D-NMR experiments along with ESIMS and HRMS analysis.The first four compounds are cyclocanthogenin and cycloastragenol glycosides, whereas the last two are based on cyclocephalogenin as aglycone, more unusual in the plant kingdom, so far reported only from Astragalus spp.     

The dinosaur tracksite of Loulle (early Kimmeridgian; Jura,France)

More than 1500 sauropod and theropod tracks have been discovered and excavated on the floor of a disused 3800 m² quarry, near the village of Loulle (French Jura). These levels correspond to a tidal-flat environment, from intertidal to supratidal zone, dated from the earliest Kimmeridgian (∼157 M.a.). Eighteen sauropod trackways have been recognized, ranging from 3.8 m to 51.5 m in length. These trackways do not correspond to a unique herd passage, because trackways are diversely oriented, most of them are crossing others and prints are more or less deeply marked, depending of different stages of substratum competence. According to the trackway gauges, the three types (narrow-, medium and wide-gauge) are represented. Some trackways could be referred to the ichnogenus Brontopodus Farlow et al., 1989, and others to Parabrontopodus Lockley et al., 1994a. Based on footprint size, the body-sizes of these 18 sauropod trackways range from tiny (Pes Length < 25 cm) to large individuals (PL > 75 cm), showing a regular continuum of sizes. Several biometrical ratios classically used in sauropod ichnology appear to be closely related to size (e.g., the smallest individuals apparently moved faster than the largest; large individuals are all narrow-gauge while small individuals are all wide-gauge; large specimens are characterized by a high heteropody ratio when the smallest show a low heteropody ratio). Associated with these sauropod trackways, four theropod trackways have also been recorded. Three small to medium-sized theropod are referred to the ichnogenus Carmelopodus Lockley et al., 1998a, while a large individual (PL = 77 cm) is tentatively referred to Megalosauripus Lessertisseur, 1955.     

A new anaxyelid wood wasp (Hymenoptera: ‘Symphyta’) from the mid-Cretaceous Burmese amber

A new genus and species of anaxyelid wasp is described and figured from the mid-Cretaceous Burmese amber as Curvitexis kopylovi gen. et sp. nov. The placement of this new genus within the Anaxyelidae is corroborated by its wing venation and the configuration of its mesosoma. This new genus differs from all other Syntexinae from the mid-Cretaceous Burmese amber biota, inter alia, owing to its forewing with the vein 1Rs curved and longer than 1M, the crossvein 1r-rs absent, the vein 2Rs + M present; the hind wing with the abscissa 2M + Cu present, the vein m-cu absent, and the cell r closed. The recently described Paraxiphydria resinata Gao et al., 2022 is transferred to Anaxyelidae: Syntexinae, and the subfamily Paraxiphydriinae Gao et al., 2022 is synonymized under Syntexinae.     

Triterpenoid saponins from Astragalus wiedemannianus Fischer

Three cycloartane-type triterpene glycosides were isolated from Astragalus wiedemannianus together with eight known secondary metabolites namely cycloastragenol, cycloascauloside B, astragaloside IV, astragaloside VIII, brachyoside B, astragaloside II, astrachrysoside A, and astrasieversianin X. The structures were established mainly by a combination of 1D and 2D-NMR techniques as 3-O-[α-L-rhamnopyranosyl-(1 → 2)-β-D-glucopyranosyl]-25-O-β-D-glucopyranosyl-20(R),24(S)-epoxy-3β,6α,16β,25-tetrahydroxycycloartane, 3-O-[α-L-rhamnopyranosyl-(1 → 2)-β-D-xylopyranosyl]-6-O-β-D-glucopyranosyl-24-O-α-(4’-O-acetoxy)-L-arabinopyranosyl-16-O-acetoxy-3β,6α,16β,24(S),25-pentahydroxycycloartane, 3-O-[α-L-rhamnopyranosyl-(1 → 2)-β-D-xylopyranosyl]-6-O-β-D-glucopyranosyl-24-O-α-L-arabinopyranosyl-16-O-acetoxy-3β,6α,16β,24(S),25-pentahydroxycycloartane. To the best of our knowledge, the presence of an arabinose moiety on the acyclic side chain of cycloartanes is reported for the first time.     

Effects of overexpressing photosynthetic carbon flux control enzymes in the cyanobacterium Synechocystis PCC 6803

Synechocystis PCC 6803 is a model unicellular cyanobacterium used in e.g. photosynthesis and CO₂ assimilation research. In the present study we examined the effects of overexpressing Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO), sedoheptulose 1,7-biphosphatase (SBPase), fructose-bisphosphate aldolase (FBA) and transketolase (TK), confirmed carbon flux control enzymes of the Calvin-Bassham-Benson (CBB) cycle in higher plants, in Synechocystis PCC 6803. Overexpressing RuBisCO, SBPase and FBA resulted in increased in vivo oxygen evolution (maximal 115%), growth rate and biomass accumulation (maximal 52%) under 100 μmol photons m⁻² s⁻¹ light condition. Cells overexpressing TK showed a chlorotic phenotype but increased biomass by approximately 42% under 100 μmol photons m⁻² s⁻¹ light condition. Under 15 μmol photons m⁻² s⁻¹ light condition, cells overexpressing TK showed enhanced in vivo oxygen evolution. This study demonstrates increased growth and biomass accumulation when overexpressing selected enzymes of the CBB cycle. RuBisCO, SBPase, FBA and TK are identified as four potential targets to improve growth and subsequently also yield of valuable products from Synechocystis PCC 6803.     

Steroidal saponins from Tribulus terrestris

Five new steroidal saponins were isolated from the fruits of Tribulus terrestris. Their structures were fully established by spectroscopic and chemical analysis as (23S,25S)-5α-spirostane-24-one-3β,23-diol-3-O-{α-l-rhamnopyranosyl-(1 → 2)-O-[β-d-glucopyranosyl-(1 → 4)]-β-d-galactopyranoside} (1), (24S,25S)-5α-spirostane-3β,24-diol-3-O-{α-l-rhamnopyranosyl-(1 → 2)-O-[β-d-glucopyranosyl-(1 → 4)]-β-d-galactopyranoside} (2), 26-O-β-d-glucopyranosyl-(25R)-5α-furostan-2α,3β,22α,26-tetraol-3-O-{β-d-glucopyranosyl-(1 → 2)-O-β-d-glucopyranosyl-(1 → 4)-β-d-galactopyranoside} (3), 26-O-β-d-glucopyranosyl-(25R)-5α-furostan-20(22)-en-2α,3β,26-triol-3-O-{β-d-glucopyranosyl-(1 → 2)-O-β-d-glucopyranosyl-(1 → 4)-β-d-galactopyranoside} (4), and 26-O-β-d-glucopyranosyl-(25S)-5α-furostan-12-one-22-methoxy-3β,26-diol-3-O-{α-l-rhamnopyranosyl-(1 → 2)-O-[β-d-glucopyranosyl-(1 → 4)]-β-d-galactopyranoside} (5). The isolated compounds were evaluated for cytostatic activity against HL-60 cells.     

A centrifugal ultrafiltration strategy for isolating the low-molecular weight (≤ 25 K) component of human plasma proteome

The low-molecular weight fraction (LMF) of the human plasma proteome is an invaluable source of biological information, especially in the context of identifying plasma-based biomarkers of disease. In this study, a separation and enrichment strategy based on centrifugal ultrafiltration was developed for the LMF (i.e., ≤ 25 K) of plasma routinely prepared from normal, healthy volunteers. Four commercially-available filter membranes of similar nominal molecular weight cut-off (NMWC), but differing membrane chemistries and filter orientations (Microcon®, Millipore; Centrisart®, Sartorius; Amicon Ultra®, Millipore; Vivaspin®, Sartorius), were evaluated. Of these filtration devices, only the Sartorius Vivaspin® tangential membrane, NMWC 20 K was effective in the non-retention of M_r > 50 K, and recovery and enrichment of low-M_r components from human plasma. This filter membrane device was further optimized with respect to plasma buffer composition, centrifugal force, duration and temperature. Optimal ultrafiltration conditions were obtained using 100 µL of normal plasma in 10% acetonitrile, and a centrifugation force of 4000 × g for 35 min at 20 °C. In this LMF, 44 proteins (from 266 unique peptides) were identified using a combination of 1D-SDS-PAGE / nano-LC-MS/MS and a stringent level of identification (FDR < 1%). We report the identification of several proteins (e.g., protein KIAA0649 (Q9Y4D3), rheumatoid factor D5, serine protease inhibitor A3, and transmembrane adapter protein PAG) previously not reported in extant high-confidence Human Proteome Organization (HUPO) Plasma Proteome Project datasets. When compared with the low-M_r human plasma/serum proteome datasets of Zhou et al. (Electrophoresis, 2004. 25, 1289–98), Gundry et al. (Proteomics Clin. Appl., 2007. 1, 73–88) and Villanueva et al. (Anal Chem, 2004. 76, 1560–70), 64% of our identifications (28 proteins) were novel; these include cofilin-1, PPIase A, and the SH3 domain-binding glutamic acid-rich-like protein 3. In addition to intact proteins, many peptide fragments from high-abundance proteins (e.g., fibrinogen, clusterin, Factor XIIIa, transferrin, kinogen-1, and inter-alpha-trypsin inhibitor), presumably derived by ex vivo proteolysis, were observed.     

Synthesis,biological evaluation,and physicochemical property assessment of 4-substituted 2-phenylaminoquinazolines as Mer tyrosine kinase inhibitors

Current results identified 4-substituted 2-phenylaminoquinazoline compounds as novel Mer tyrosine kinase (Mer TK) inhibitors with a new scaffold. Twenty-one 2,4-disubstituted quinazolines (series 4–7) were designed, synthesized, and evaluated against Mer TK and a panel of human tumor cell lines aimed at exploring new Mer TK inhibitors as novel potential antitumor agents. A new lead, 4b, was discovered with a good balance between high potency (IC₅₀ 0.68 μM) in the Mer TK assay and antiproliferative activity against MV4-11 (GI₅₀ 8.54 μM), as well as other human tumor cell lines (GI₅₀ < 20 μM), and a desirable druglike property profile with low log P value (2.54) and high aqueous solubility (95.6 μg/mL). Molecular modeling elucidated an expected binding mode of 4b with Mer TK and necessary interactions between them, thus supporting the hypothesis that Mer TK might be a biologic target of this kind of new active compound.     

Optimisation of preparative HPLC separation of four isomeric kaempferol diglycosides from Prunus spinosa L. by application of the response surface methodology

A fast and efficient preparative HPLC-PDA method was developed for the separation and isolation of four rare isomeric kaempferol diglycosides from leaves of Prunus spinosa L. The separation procedure of the enriched diglycoside fraction of the 70% (v/v) aqueous methanolic leaf extract was first optimised on analytical XBridge C18 column (100 mm × 4.6 mm i.d., 5 μm) and central composite design combined with response surface methodology was utilized to establish the optimal separation conditions. The developed method was directly transferred to preparative XBridge Prep C18 column (100 mm × 19 mm i.d., 5 μm) and the final separation was accomplished by isocratic elution with 0.5% acetic acid-methanol-tetrahydrofuran (75.2:16.6:8.2, v/v/v) as the mobile phase, at a flow rate of 13.6 mL/min, in less than 12 min for a single run. Under these conditions, four flavonoid diglycosides: kaempferol 3-O-α-l-arabinofuranoside-7-O-α-l-rhamnopyranoside, kaempferol 3,7-di-O-α-l-rhamnopyranoside (kaempferitrin), and reported for the first time for P. spinosa kaempferol 3-O-β-d-xylopyranoside-7-O-α-l-rhamnopyranoside (lepidoside) and kaempferol 3-O-α-l-arabinopyranoside-7-O-α-l-rhamnopyranoside, were isolated in high separation yield (84.8–94.5%) and purity (92.45–99.79%). Their structures were confirmed by extensive 1D and 2D NMR studies. Additionally, the UHPLC-PDA-ESI–MS³ qualitative profiling led to the identification of twenty-one phenolic compounds and confirmed that the isolates were the major components of the leaf material.     

Morphology and SSU rRNA gene sequences of three Frontonia species,including a description of F. subtropica spec. nov. (Ciliophora,Peniculida)

The morphology and infraciliature of three Frontonia species, F. subtropica spec. nov., F. canadensis Roque and Puytorac, 1972, and F. magna Fan et al., 2011, isolated from coastal waters in southern China sea, were investigated using living observation and silver impregnation methods. Frontonia subtropica spec. nov. is recognized by the combination of the following characters: body elliptical in outline with right margin depressed in anterior third, about 180–230 μm × 60–80 μm in vivo; 104–114 somatic kineties; peniculi 1–3 each with four kineties; five vestibular and five postoral kineties; one centrally located elongate-elliptical macronucleus; single contractile vacuole located left-dorsally in posterior third of body. We also provide improved diagnoses for F. canadensis and F. magna based on current and previous reports. The small subunit (SSU) rRNA gene was sequenced for all three species. Comparisons with sequences of morphologically similar congeners clearly support the validity each species.     

Retrieval of phycocyanin concentration from remote-sensing reflectance using a semi-analytic model in eutrophic lakes

With the rapid development of the economy in recent years, massive algal (blue-green algae in particular) blooms have often observed in Chinese eutrophic lakes. The concentration of the cyanobacterial pigment phycocyanin (PC), an accessory pigment unique to freshwater blue-green algae, is often used as a quantitative indicator of blue-green algae in eutrophic inland waters. The purpose of this study was to evaluate the semi-analytic PC retrieval algorithm proposed by Simis et al. and to explore the potential to improve this PC algorithm so that it is more suitable for eutrophic lakes, such as Taihu Lake. In this paper, we recalculated the correction coefficients γ and δ to calculate the absorptions of chlorophyll-a at 665 nm and the absorptions of phycocyanin at 620 nm in terms of in situ measurements and observed that the values of these coefficients differed from the values used by Simis et al. and Randolph et al. The two coefficients are site dependent due to the different bio-optical properties of lakes. We also observed that the specific PC absorption at 620 nm a_pc*(620) decreases exponentially with an increase in PC concentrations. Therefore, a non-linear power–function of a_pc*(620), instead of a constant value of a_pc*(620) as used by Simis et al., was proposed for our improved PC retrieval algorithm in Taihu Lake, yielding a squared correlation coefficient (R²) of 0.55 and a root mean square error (RMSE) of 58.89 μg/L. Compared with the original PC retrieval algorithm by Simis et al., the improved retrieval algorithm has generally superior performance. In evaluating the limitation of the PC retrieval algorithms, we observed that the ratio of the total suspended solids to phycocyanin can be used as a primary measure for retrieval performance. Validation in Dianchi Lake and an error analysis proved that the improved PC algorithm has a better universality and is more suitable for eutrophic lakes with higher PC concentrations.     

Chemical constituents from Tribulus terrestris and screening of their antioxidant activity

Two oligosaccharides (1, 2) and a stereoisomer of di-p-coumaroylquinic acid (3) were isolated from the aerial parts of Tribulus terrestris along with five known compounds (4–8). The structures of the compounds were established as O-β-d-fructofuranosyl-(2 → 6)-α-d-glucopyranosyl-(1 → 6)-β-d-fructofuranosyl-(2 → 6)-β-d-fructofuranosyl-(2 → 1)-α-d-glucopyranosyl-(6 → 2)-β-d-fructofuranoside (1), O-α-d-glucopyranosyl-(1 → 4)-α-d-glucopyranosyl-(1 → 4)-α-d-glucopyranosyl-(1 → 2)-β-d-fructofuranoside (2), 4,5-di-p-cis-coumaroylquinic acid (3) by different spectroscopic methods including 1D NMR (¹H, ¹³C and DEPT) and 2D NMR (COSY, TOCSY, HMQC and HMBC) experiments as well as ESI-MS analysis. This is the first report for the complete NMR spectral data of the known 4,5-di-p-trans-coumaroylquinic acid (4).The antioxidant activity represented as DPPH free radical scavenging activity was investigated revealing that the di-p-coumaroylquinic acid derivatives possess potent antioxidant activity so considered the major constituents contributing to the antioxidant effect of the plant.     

Membrane-like protein involved in phage adsorption associated with phage-sensitivity in the bloom-forming cyanobacterium Microcystis aeruginosa

The cyanobacterium, Microcystis aeruginosa, contains a large number of defense genes (Makarova et al., 2011); thus, it is a good model to study the co-evolution of phage and bacteria. Here, we isolated and characterized two phage-resistant M. aeruginosa mutants that came from a phage intermediate-sensitive culture. To determine the mutation conferring resistance, a protein expression pattern analysis was performed comparing phage-sensitive and -resistant sub-strains using SDS-PAGE. There were no apparent differences in expression patterns in the soluble fraction; however, a ∼90 kDa protein in the hydrophobic fraction from the phage-sensitive sub-strain was observed. Using a successive thermal asymmetric interlaced-PCR, the entire sequence encoding the protein, assigned ISP90, as well as its neighboring regions (ca. 7.8 kb) was determined. ISP90 contained no conserved domains and was predicted to be a membrane-associated protein. No mutations were detected in the nucleotide sequences coding ISP90 and diversification of ISP90 regions within this species were observed. Diversification of ISP90 regions within this species suggests a possible genomic island that may be subjected to selective pressures from phages. The ISP90 sequence involving phage resistance/sensitivity contributes to the understanding of co-evolution between M. aeruginosa and phages.     

Highly glycosylated flavonols with an O-linked branched pentasaccharide from Iberis saxatilis (Brassicaceae)

Four flavonol glycosides isolated from non-flowering leafy shoots of Iberis saxatilis (Brassicaceae) were characterised by spectroscopic and chemical methods as saxatilisins A–D, the 3-O-β-d-glucopyranosyl-(1 → 3)-α-l-rhamnopyranosyl-(1 → 2)[β-d-glucopyranosyl-(1 → 2)-α-l-rhamnopyranosyl-(1 → 6)]-β-d-glucopyranoside, 3-O-β-d-glucopyranosyl-(1 → 3)-α-l-rhamnopyranosyl-(1 → 2)[α-l-rhamnopyranosyl-(1 → 6)]-β-d-glucopyranoside, 3-O-(6-O-E-sinapoyl)-β-d-glucopyranosyl-(1 → 3)-α-l-rhamnopyranosyl-(1 → 2)[β-d-glucopyranosyl-(1 → 2)-α-l-rhamnopyranosyl-(1 → 6)]-β-d-glucopyranoside, and 3-O-(6-O-E-feruloyl)-β-d-glucopyranosyl-(1 → 3)-α-l-rhamnopyranosyl-(1 → 2)[β-d-glucopyranosyl-(1 → 2)-α-l-rhamnopyranosyl-(1 → 6)]-β-d-glucopyranoside of isorhamnetin (3,5,7,4′-tetrahydroxy-3′-methoxyflavone), respectively. Analysis of ²J_HC correlations detected with the H2BC (heteronuclear two-bond correlation) pulse sequence aided the unambiguous assignment of glycosidic resonances in the ¹H and ¹³C NMR spectra of these compounds. Saxatilisins A, C, and D, are the first flavonol glycosides to be described with a pentasaccharide chain at a single glycosylation site. Several pentaglycosides of kaempferol and quercetin, tentatively assigned as saxatilisin analogues from LC–MS/MS analyses, were present as minor constituents of the extracts.     

Comparaison de six méthodes de segmentation du volume tumoral sur la 18F-FDG TEP-TDM avec le volume de référence anatomopathologique dans les cancers bronchopulmonaires non à petites cellules

The ¹⁸F-FDG TEP has demonstrated its importance in oncology, for initial extension and efficacy of antitumoral therapeutics. Several studies have attempted to prove its utility to define tumoral volumes for conformational radiotherapy in nonsmall cell lung cancers. Some authors have suggested the use of threshold of tumor intensity uptake with 40 or 50% of maximal intensity. Black et al. have determined contouring with linear regression formula of mean semi-quantitative index of tumor uptake (standard uptake value): SUV_threshold = 0.307 SUV_average + 0.588. Nestlé et al. have taken into account the background noise intensity and mean intensity of the tumor: I_threshold = β I_average + I_noise with β = 0.15. Our study was done in collaboration with Inserm U618 team and has compared volumes defined on PETscan defined according to different methods based on intensity or SUV to the tumour volume determined on CT scan by radiophysicist. We have compared those volumes with histological volume that we considered for reference. Four patients have been included. They had ¹⁸F-FDG PETscan followed by complete tumoral removal surgery. Specific histological procedure allowed to define complete size of the tumor in reexpanded lung. Comparatively to pathology, the volumes obtained using I_max 40 and I_max 50 are all underestimated. The volumes defined by Black's et al. method are underevaluated for the two largest tumours (15.8% to 22%) and overestimated for the two smallest ones (17.9 to 82.9%). Nestlé's et al. method, using β = 0.15, correctly estimates two tumor volumes over 2 cm, but overestimates the two small tumors (79.6 to 124%). Finally, the corrected Nestlé's et al. formula (using β = 0.264) overestimates three tumours. Volumes defined on CT scan by radiophysicist are correct for one lesion, underestimated for one and overestimated for two other ones (44 and 179.5%). Nestlé's et al. method seems to be the most accurate for tumours over 2 cm of diameter. Today no method can correctly estimate the volume of smaller tumours. To improve those segmentation approaches, partial volume effect correction needs to be implemented for small tumours or tumours close to mediastinal local extension. The main indication of segmentation based on FDG PET is tumour greater than 2 cm for which treatment will be radiotherapy alone or radiotherapy associated with chemotherapy. Our study is important because it compares the different methods described in the literature with the reference histological volume of the tumor, which is the only way to validate tumor segmentation method.