Effect of age,size and digestive tract development on weaning effectiveness in crucian carp,Carassius carassius (Linnaeus, 1758)

The study aim was to determine the optimum age, wet body weight (WBW) and total length (TL) of the crucian carp, Carassius carassius (L.), to ensure the effectiveness of weaning directly without a gradual transfer from live food to a compound feed. Moreover, the state of development of the digestive tract was analyzed histologically based on the height of enterocytes. Experimental rearing was conducted between days 5 and 45 post hatch (DPH). Initial WBW of fish was 2.2 ± 0.6 (n = 30) mg and TL 6.1 ± 0.1 (n = 30) mm. Rearing was carried out at 27 ± 0.5°C, with fish divided into six groups: one control (C) fed with Artemia sp. nauplii, and five groups initially fed with Artemia sp. but later replaced by a compound feed. Weaning with the compound diet started at 15, 20, 25, 30 and 35 DPH in groups labeled F15, F20, F25, F30, F35, respectively. Larvae were fed three times per day (08.00 h, 13.00 h, 18.00 h) in equal portions (4% of larvae biomass per day, converted to the dry matter of the feed). Daily biomass growth was adopted as 15%. Each group was triplicated (n = 50 individuals per replicate). Highest values of TL 42.1 ± 0.7 (n = 30) mm and WBW 905.3 ± 50.3 (n = 30) mg were recorded in the control group at 45 DPH; lowest survival rate of 45 DPH was in group F15 (90.7 ± 1.2%, n = 30). The highest value of the enterocyte epithelial length was observed in individuals within groups F30, 34.8 ± 1.2 μm (n = 30) and F35, 35.4 ± 3.6 μm (n = 30), respectively, 30 and 35 DPH; highest percentage of deformations on the final day of the experiment was in group F15 (100 ± 0.0%, n = 30). The results indicate that an effective direct transfer from live food to prepared diets (with no gradual transfer) cannot be performed with crucian carp larvae before 30 DPH at 27°C, when the fish have reached TL = 31.1 ± 0.4 mm (n = 30) and WBW = 436.9 ± 13.7 mg (n = 30).     

The toxicity of the Aggregatibacter actinomycetemcomitans cytolethal distending toxin correlates with its phosphatidylinositol‐3,4,5‐triphosphate phosphatase activity

The Aggregatibacter actinomycetemcomitans cytolethal distending toxin (Cdt) induces G2 arrest and apoptosis in lymphocytes and other cell types. We have shown that the active subunit, CdtB, exhibits phosphatidylinositol‐3,4,5‐triphosphate (PIP3) phosphatase activity, leading us to propose that Cdt toxicity is the result of PIP3 depletion and perturbation of phosphatidylinositol‐3‐kinase (PI‐3K)/PIP3/Akt signalling. To further explore this relationship, we have focused our analysis on identifying residues that comprise the catalytic pocket and are critical to substrate binding rather than catalysis. In this context, we have generated several CdtB mutants and demonstrate that, in each instance, the ability of the toxin to induce cell cycle arrest correlates with retention of phosphatase activity. We have also assessed the effect of Cdt on downstream components of the PI‐3K signalling pathway. In addition to depletion of intracellular concentrations of PIP3, toxin‐treated lymphocytes exhibit decreases in pAkt and pGSK3β. Further analysis indicates that toxin‐treated cells exhibit a concomitant loss in Akt activity and increase in GSK3β kinase activity consistent with observed changes in their phosphorylation status. We demonstrate that cell susceptibility to Cdt is dependent upon dephosphorylation and concomitant activation of GSK3β. Finally, we demonstrate that, in addition to lymphocytes, HeLa cells exposed to a CdtB mutant that retains phosphatase activity and not DNase activity undergo G2 arrest in the absence of H2AX phosphorylation. Our results provide further insight into the mode of action by which Cdt may function as an immunotoxin and induce cell cycle arrest in target cells such as lymphocytes.     

The predictive value of sonographic images of follicular lesions – a comparison with nodules unequivocal in FNA – single centre prospective study

Background

To determine the diagnostic efficacy of ultrasonographic malignancy risk features (UMRFs) in follicular lesions (FL) in a population with low risk of malignancy in FL and to compare it with a similar analysis in a group of patients with unequivocal cytology (UC): benign lesion (BL) or malignant neoplasm (MN).

Methods

Presence of UMRFs (hypoechogenicity, solid echostructure, taller-than-wide shape, pathological vascularization, irregular margins, microcalcifications and macrocalcifications) and their sets were assessed in 322 FL: 202 follicular lesions of undetermined significance (FLUS) and 120 suspicious for follicular neoplasm (SFN) and 300 nodules with UC: 200 BL and 100 MN, subsequently evaluated histopathologically.

Results

Cancers were confirmed in 100% nodules in MN group (89.0% of them were papillary carcinomas - PTC), in 6.4% FLUS nodules (69.2% PTC), and in 10.8% SFN nodules (30.8% PTC). In the UC group all UMRFs occurred more frequently in cancers than in benign lesions. In the FL group only calcifications were found in cancers more frequently – macro and microcalcifications together: 34.6 vs. 11.5% (p?=?0.001) and isolated macrocalcifications: 26.0 vs. 6.8% (p?=?0.001); the presence of those features increased the basic risk of malignancy in FL more than 2 times. The presence of at least 2 of the following URMFs: hypoechogenicity, solid echostructure, any type of calcifications and suspected shape, additionally improved sensitivity.

Conclusions

Evaluation of UMRFs in FLs is less effective than in nodules with UC, and its effectiveness decreases parallel to the decrease in percentage of PTCs among malignant neoplasms and to the increase of the percentage of adenomas among benign nodules. The presence of macrocalcifications in such FLs significantly increases the basic risk of malignancy in these nodules.

     

Identification and inhibition of carbonic anhydrases from nematodes

Abstract

Carbonic anhydrases (CAs) are metalloenzymes, and classified into the evolutionarily distinct α, β, γ, δ, ζ, and η classes. α-CAs are present in many living organisms. β- and γ-CAs are expressed in most prokaryotes and eukaryotes, except for vertebrates. δ- and ζ-CAs are present in phytoplanktons, and η-CAs have been found in Plasmodium spp. Since the identification of α- and β-CAs in Caenorhabditis elegans, the nematode CAs have been considered as an emerging target in research focused on antiparasitic CA inhibitors. Despite the presence of α-CAs in both helminths and vertebrates, structural studies have revealed different kinetic and inhibition results. Moreover, lack of β-CAs in vertebrates makes this enzyme as an attractive target for inhibitory studies against helminthic infection. Some CA inhibitors, such as sulfonamides, have been evaluated against nematode CAs. This review article aims to present comprehensive information about the nematode CAs and their inhibitors as potential anthelminthic drugs.     

Design,synthesis and biological evaluation of 4-benzoyl-1-dichlorobenzoylthiosemicarbazides as potent Gram-positive antibacterial agents

Twelve 4-benzoyl-1-dichlorobenzoylthiosemicarbazides have been tested as potential antibacterials. All the compounds had MICs between 0.49 and 15.63?µg/ml toward Micrococcus luteus, Bacillus cereus, Bacillus subtilis and Staphylococcus epidermidis indicating, in most cases, equipotent or even more effective action than cefuroxime. In order to clarify if the observed antibacterial effects are universal, further research were undertaken to test inhibitory potency of two most potent compounds 3 and 11 on clinical isolates of Staphylococcus aureus. Compound 11 inhibited the growth of methicillin-sensitive S. aureus (MSSA) at MICs of 1.95–7.81?µg/ml, methicillin-resistant S. aureus (MRSA) at MICs of 0.49–1.95?µg/ml and MDR–MRSA at MIC of 0.98 and 3.90?µg/ml, respectively. Finally, inhibitory efficacy of 3 and 11 on planktonic cells and biofilms formation in clinical isolates of S. aureus and Haemophilus parainfluenzae was tested. The majority of cells in biofilm populations of MSSA and MRSA were eradicated at low level of 3, with MBICs in the range of 7.82–15.63?µg/ml.     

Synthesis and anticancer and lipophilic properties of 10-dialkylaminobutynyl derivatives of 1,8- and 2,7-diazaphenothiazines

New derivatives of two isomeric types of azaphenothiazines, 1,8- and 2,7-diazaphenothiazine, containing the triple bond substituents and additionally tertiary cyclic and acyclic amine groups, were synthesized and tested for their anticancer activity. The compounds exhibited differential inhibitory activities. Better results were obtained when the acetylenic group was transformed via the Mannich reaction to the dialkylaminobutynyl groups. The most active was 2,7-diazaphenothiazine with the N-methylpiperazine-2-butynyl substituent against the human ductal breast epithelial tumor cell line T47D, more potent than cisplatin. The 2,7-diazaphenothiazine system turned out to be more active than isomeric 1,8-diaza one. For the most active compound, the expression of TP53, CDKN1A, BCL-2 and BAX genes was detected by the RT-QPCR method. The gene expression ratio BACL-2/BAX suggests the mitochondrial apoptosis in T47D cells. The synthesis makes possible to obtain many new bioactive phenothiazines with the dialkylaminoalkynyl substituents inserting various tertiary cyclic and acyclic amine moieties to the substituents.     

The effect of dietary methionine levels on fattening performance and selected blood and tissue parameters of turkeys

A total of 490 eight-week-old female Hybrid Converter turkeys (body weight 4.11 ± 0.03 kg) were divided into 5 groups with 7 replicates of 14 birds each. For 8 weeks, basal diets were supplemented with methionine (Met) at following levels (weeks 9–12/weeks 13–16 of age): Group 1 – 0.34/0.29%, Group 2 – 0.39/0.34%, Groups 3 and 4 – 0.45/0.38% and 0.51/0.41%, respectively, Group 5 – 0.58/0.47%. Only in the first feeding phase the body weight gain (BWG) was affected by Met levels with the significantly highest BWG in Group 3. No treatment effects were found for feed conversion ratio, carcass yield, carcass composition and meat colour. The blood superoxide dismutase activity was significantly highest in Groups 2 and 3. The concentrations of reduced glutathione in the liver were linearly increased (p = 0.018), whereas the ratio of reduced glutathione to oxidised glutathione was highest in Group 3 (quadratic contrast, p = 0.004). It can be concluded that turkeys from Group 3 (Met levels age depending 15% and 10% above recommendations by NRC) were characterised by a well-balanced physiological response. Attention should be paid to the immune response of birds to higher dietary Met levels: plasma IgA concentrations decreased, whereas IL-6 and TNF-α levels increased in turkeys fed diets with the highest Met content.