L-amino acid oxidase from snake venom: Biotransformation and induction of apoptosis in human colon cancer cells

Chronic periodontitis is an inflammatory disease that represents a major public health issue nowadays. Here, we investigated the protective role of NF-κB inducing kinase (NIK)-inhibitor on chronic periodontitis and revealed the underlying molecular mechanism. NIK-inhibitor was synthesized, and its functions were examined in primary osteoclasts and wild-type (WT) and NIK^-/- chronic periodontitis mouse model. Lipopolysaccharides (LPS) or activator of NF-κB was applied to stimulate inflammatory response of osteoclasts.

The qRT-PCR, ELISA and Western blot were used to measure the expression of pro-inflammatory and osteoclast-related genes, and the activation of NF-κB signaling. Osteoclastogenesis and bone damage were detected by TRAP staining and micro-CT. NIK knockdown mice had lower expression of osteoclast-related genes and improved CEJ-ABC damage.

Similarly, NIK-inhibitor administration inhibited inflammatory responses and CEJ-ABC damage in chronic periodontitis models. NIK-inhibitor suppressed osteoclastogenesis and osteoclast-related genes expression through inhibiting the non-canonical NF-κB signaling. NIK plays important role in bone destruction of chronic periodontitis and NIK-inhibitor represents a promising therapeutic strategy for this disease.

The complexity of the activated B-cell like (ABC) diffuse large B-cell lymphoma (DLBCL) subtype is probably not only explained by genetic alterations and methods to measure global protein expression could bring new knowledge regarding the pathophysiology. We used quantitative proteomics to analyze the global protein expression of formalin-fixed paraffin-embedded (FFPE) tumor tissues from 202 DLBCL patients.

We identified 6430 proteins and 498 were significantly regulated between the germinal center B-cell like (GCB) and non-GCB groups. A number of proteins previously not described to be upregulated in non-GCB or ABC DLBCL was found, e.g. CD64, CD85A, guanylate-binding protein 1 (GBP1), interferon-induced proteins with tetratricopeptide repeat (IFIT)2, and mixed lineage kinase domain-like protein (MLKL) and immunohistochemical staining showed higher expression of GBP1 and MLKL.

A cluster analysis revealed that the most prominent cluster contained proteins involved in the tumor microenvironment and regulation of the immune system. Our data suggest that the therapeutic focus should be expanded toward the tumor microenvironment in non-GCB/ABC subtype patients.

The ability of Listeria monocytogenes, an important foodborne pathogen, to form biofilms in food processing environments leads to increased opportunity for contamination of food products, which is a major concern for food safety. In this study, the role of a complex system composed of the VirSR two-component signal transduction system (TCS) and the ATP-binding cassette (ABC) transporter VirAB in biofilm formation of L. monocytogenes EGD-e was investigated.

Biofilm formation was measured using the microplate assay with crystal violet staining, confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM), and attachment and swarming motility were compared between strain EGD-e and its isogenic deletion mutants. Additionally, the relative expression levels of genes associated with the early steps of biofilm development in the wild-type and mutant strains were also determined by RT-qPCR. Results from microplate assay, CLSM and SEM showed that VirR is not required for biofilm formation in L. monocytogenes EGD-e.

A central finding of this study is that both VirAB and VirS are essential for biofilm formation and they could function as a whole in biofilm formation of L. monocytogenes EGD-e. The results also demonstrated that both VirAB and VirS are involved in attachment, but they are not associated with swarming motility. Results from RT-qPCR showed that flaA, motA and motB were downregulated in the mutant strains ΔvirAB and ΔvirS, which could be the possible reason for reduced attachment and biofilm formation in these mutants. This study provides a better understanding of the mechanisms involved in biofilm formation of L. monocytogenes, leading to improved processes to control this biofilm-forming foodborne pathogen.

In a raw-meat-processing environment, members of the Enterobacteriaceae family can coexist with Staphylococcus aureus to form dual-species biofilms, leading to a higher risk of food contamination. However, very little is known about the effect of inter-species interactions on dual-species biofilm formation.

The aim of this study was to investigate the interactions between S. aureus and raw-meat-processing environment isolates of Klebsiella oxytoca in dual-species biofilms, by employing an untargeted metabolomics tool. Crystal violet staining assay showed that the biomass of the dual-species biofilm significantly increased and reached its maximum after incubation for 21 h, compared with that of single species grown alone.

The number of K. oxytoca in the dual-species biofilm was significantly higher than that of S. aureus. Field emission scanning electron microscopy (FESEM) revealed that both species were evenly distributed, and were tightly wrapped by extracellular polymeric substances in the dual-species biofilms. Ultra-high-pressure liquid chromatography equipped with a quadrupole-time-of-flight mass spectrometer (UHPLC-Q-TOF MS) analysis exhibited a total of 8184 positive ions, and 6294 negative ions were obtained from all test samples.

Ponceau S Staining Solution
B7778-100000	ApexBio	100 g	EUR 195.6

Ponceau S Staining Solution
B7778-50000	ApexBio	50 g	EUR 151.2

SDS-PAGE Staining Solution
GR103068	Genorise Scientific	100 mL	EUR 58

Ponceau S Staining Solution
PW005	Bio Basic	100ml	EUR 82.97

Dual- Color AP Staining Kit
AP100D-1	SBI	100 Assays	EUR 494.4

EasyDip Slide Staining System
M900-12AS-ASSORTED	ScyTek Laboratories	1 Case(s)	EUR 211.2

Red-Color AP Staining Kit
AP100R-1	SBI	50 Assays	EUR 314.4

Live/Dead Cell Staining Kit
55R-1434	Fitzgerald	100 tests	EUR 765.6
Description: Live/Dead Cell Staining Kit for use in the research laboratory

PAP PEN FOR IMMUNO STAINING
Z377821-1EA	Scientific Laboratory Supplies	EACH	EUR 171.03

Stain Tray Slide Staining System
M920-1	ScyTek Laboratories	1 Clear	EUR 513.6

Stain Tray Slide Staining System
M920-2	ScyTek Laboratories	1 Black	EUR 534

CometAssay Silver Staining Kit
4254-200-K	Biotechne	200 Tests	EUR 390.96

Nalgene Box Staining 13x13x5cm
BOX1040	Scientific Laboratory Supplies	EACH	EUR 48

Blue-Color AP Staining Kit
AP100B-1	SBI	50 Assays	EUR 203

Beta Galactosidase Staining Kit
55R-1541	Fitzgerald	250 assays	EUR 444
Description: Assay Kit for detection of Beta Galactosidase in the research laboratory

Disposable Slide Staining Trays
MIC5070	Scientific Laboratory Supplies	PK4	EUR 90

Staining Trough+Lid Rectangular
MIC6008	Scientific Laboratory Supplies	EACH	EUR 27.6

ProsiBlue Gel Staining Solution
P7300-050	GenDepot	500ml	EUR 208.8

Live-Dead Cell Staining Kit
K501-100	Biovision	each	EUR 561.6

Live-Dead Cell Staining Kit
K2081-100	ApexBio	100 staining	EUR 552

Wheaton staining dishes 3 EA
S5017-3EA	Scientific Laboratory Supplies	PK3	EUR 135.6

Staining Jar PP + Lid 10 Slide
MIC6002	Scientific Laboratory Supplies	EACH	EUR 67.2

Purple-Color AP Staining Kit
AP100P-1	SBI	100 Assays	EUR 235

Nerve Terminal Staining Kit V
70034	Biotium	1SET	EUR 610.8
Description: Minimum order quantity: 1 unit of 1SET

Human Histochemistry Staining Kit
20-abx299039	Abbexa	EUR 1045.20 EUR 1462.80 EUR 678.00	150 tests 500 tests 50 tests

Beta-Galactosidase Staining Kit
K802-250	Biovision	each	EUR 352.8

Alkaline Phosphatase Staining Kit
K2035-50	Biovision	50 assays	EUR 454.8

Beta-Galactosidase Staining Kit
K2182-250	ApexBio	250 assays	EUR 351.6

Lipid (Oil Red O) staining Kit
K580-24	Biovision	each	EUR 542.4

Staining Jar W Lid 115x115x77mm
MIC6016	Scientific Laboratory Supplies	PK10	EUR 428.4

Manual Staining System (12 Wells)
IMS001	ScyTek Laboratories	1 ea.	EUR 427.2

Red Active Caspase Staining Kit
K2052-100	ApexBio	100 assays	EUR 686.4

Red Active Caspase Staining Kit
K2052-25	ApexBio	25 assays	EUR 285.6

Alcian Blue (pH 2.5) Staining Kit
AFR-1	ScyTek Laboratories	250 ml ea.	EUR 249.6

Alcian Blue (pH 2.5) Staining Kit
AFR-2	ScyTek Laboratories	30 ml ea.	EUR 153.6

DNA Ploidy Analysis Staining Kit
K1439-1	Biovision	each	EUR 705.6

DNA Ploidy Analysis Staining Kit
K1439-5	Biovision	each	EUR 2402.4

Staining Dish with Cover Plastic
X24085	Scientific Laboratory Supplies	EACH	EUR 156

7-AAD Viability Staining Buffer
abx090617-100tests	Abbexa	100 tests	EUR 260.4

Simport EasyDip Staining Jar Blue
HIS0273	Scientific Laboratory Supplies	PK6	EUR 117.6

Simport EasyDip Staining Jar Pink
HIS0275	Scientific Laboratory Supplies	PK6	EUR 117.6

Stayrightâ„¢ Purple HRP Staining Kit
45905-5mL	AAT Bioquest	5 mL	EUR 109
Description: 3,3'-Diaminobenzidine (DAB) has been applied for decades as the most commonly used IHC chromogen because it is inexpensive and sensitive for routine applications.

Stayrightâ„¢ Purple HRP Staining Kit
45906-50mL	AAT Bioquest	50 mL	EUR 334
Description: 3,3'-Diaminobenzidine (DAB) has been applied for decades as the most commonly used IHC chromogen because it is inexpensive and sensitive for routine applications.

Simport EasyDip Staining Jar Green
HIS0274	Scientific Laboratory Supplies	PK6	EUR 117.6

Multivariate data analysis further described altered metabolic profiling between mono- and dual-species biofilms. Further, 18 and 21 different metabolites in the dual-species biofilm were screened as biomarkers by comparing the mono-species biofilms of S. aureus and K. oxytoca, respectively. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways that were exclusively upregulated in the dual-species biofilm included ABC transporters, amino acid metabolism, and the two-component signal transduction system. Our results contribute to a better understanding of the interactive behavior of inter-species biofilm communities, by discovering altered metabolic profiling.