Journal: BMC Microbiology

Article Title: Real-time quantitative PCR assay development and application for assessment of agricultural surface water and various fecal matter for prevalence of Aliarcobacter faecis and Aliarcobacter lanthieri

doi: 10.1186/s12866-020-01826-3

Figure Lengend Snippet: List of reference strains of target and other bacterial species and strains used in this study

Article Snippet: 66 , S. epidermidis , Clinical isolate , ATCC 12228.

Techniques: Infection

Journal: Allergy

Article Title: Staphylococcus aureus causes aberrant epidermal lipid composition and skin barrier dysfunction

doi: 10.1111/all.15640

Figure Lengend Snippet: Expression of ELOVLs in human primary keratinocytes and organotypic skin cultures stimulated with Staphylococci . Gene expression of ELOVLs in organotypic skin cultures stimulated with S . epidermidis (A), MSSA (B), and MRSA (C). Data are representative of three independent experimental repetitions using three different lots of HEKs S . epidermidis , MSSA, and MRSA, respectively. n = 9 per group. Immunostaining intensities of ELOVL3 and ELOVL4 in organotypic skin cultures stimulated with MRSA (D). Arrows point to ELOVL3 and ELOVL4 staining (red). Wheat germ agglutinin-conjugated FITC (green) was used to stain the cytoskeleton. DAPI stained nuclei (blue). n = 3. Scale bar = 50 μm. All data are shown as the median and interquartile range. * p < .05, ** p < .01, *** p < .001 by the Kruskal–Wallis test with the post hoc test. ELOVL, elongation of very long chain fatty acids protein; MFI, mean fluorescence intensity; MRSA, methicillin-resistant Staphylococcus aureus ; MSSA, methicillin-sensitive Staphylococcus aureus.

Article Snippet: Then, cells were stimulated with three strains of live S . epidermidis (ATCC ® -12228 TM , ATCC ® -49461 TM , and ATCC ® -700296 TM , Manassas, VA, 0.001 or 0.01 CFU/well), three strains of methicillin-sensitive S . aureus (MSSA) (ATCC ® -29213 TM , ATCC ® -49230 TM , and SMC1801–45, Asian Bacterial Bank, Seoul Korea, 0.001 or 0.01 CFU/well), or three strains of MRSA (ATCC ® -BAA-1556 TM , ATCC-700699 TM , and SMC1803–07, Asian Bacterial Bank, 0.001 or 0.01 CFU/well) for an additional 24 h, prior to RNA collection.

Techniques: Expressing, Gene Expression, Immunostaining, Staining, Fluorescence

Journal: Allergy

Article Title: Staphylococcus aureus causes aberrant epidermal lipid composition and skin barrier dysfunction

doi: 10.1111/all.15640

Figure Lengend Snippet: Expression of keratinocyte-derived cytokines in human primary keratinocytes stimulated with Staphylococci . HEKs were Ca2+ differentiated and stimulated with three different strains of S . epidermidis , MSSA or MRSA for 8 or 20 h. Gene expression of keratinocyte-derived cytokines in HEKs stimulated with MSSA (A) and MRSA (B) was evaluated by RT-PCR. n = 27 per group. Protein levels of TNF-α (C) and IL-1β (D) in HEKs culture media were measured by ELISA. n = 4 per group. Data are representative of three independent experimental repetitions using three different lots of HEKs. All data are shown as the median and interquartile range. * p < .05, ** p < .01, *** p < .001, **** p < .0001 by the Kruskal–Wallis test with the post hoc test. MRSA, methicillin-resistant Staphylococcus aureus ; MSSA, methicillin-sensitive Staphylococcus aureus ; RT-PCR, reverse transcription-polymerase chain reaction.

Article Snippet: Then, cells were stimulated with three strains of live S . epidermidis (ATCC ® -12228 TM , ATCC ® -49461 TM , and ATCC ® -700296 TM , Manassas, VA, 0.001 or 0.01 CFU/well), three strains of methicillin-sensitive S . aureus (MSSA) (ATCC ® -29213 TM , ATCC ® -49230 TM , and SMC1801–45, Asian Bacterial Bank, Seoul Korea, 0.001 or 0.01 CFU/well), or three strains of MRSA (ATCC ® -BAA-1556 TM , ATCC-700699 TM , and SMC1803–07, Asian Bacterial Bank, 0.001 or 0.01 CFU/well) for an additional 24 h, prior to RNA collection.

Techniques: Expressing, Derivative Assay, Gene Expression, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Reverse Transcription, Polymerase Chain Reaction

Journal: Experimental and Therapeutic Medicine

Article Title: Effects of mevalonate kinase interference on cell differentiation, apoptosis, prenylation and geranylgeranylation of human keratinocytes are attenuated by farnesyl pyrophosphate or geranylgeranyl pyrophosphate

doi: 10.3892/etm.2020.8569

Figure Lengend Snippet: The mRNA and protein expression of keratin 1 and involucrin following the interference and overexpression of MVK. (A) The mRNA expression of keratin 1 and involucrin. (B) The protein expression of keratin 1 and involucirn. Relative mRNA and protein expression of keratin 1 and involucrin were detected by RT-qPCR and western blotting, respectively. Compared with the negative control group, the mRNA and protein expression of keratin 1 and involucrin were significantly decreased following interference of MVK expression. The decrease in expression of keratin 1 and involucrin following MVK interference was notably attenuated by FPP. Data presented as mean ± SD, n=3/group. ** P<0.01 and *** P<0.001 vs. negative control group. ## P<0.01 and ### P<0.001 vs. MVK interference group. KRT1, keratin 1; IVL, involucrin; MVK, mevalonate kinase; RT-qPCR, reverse transcription-quantitative PCR; FPP, farnesyl pyrophosphate.

Article Snippet: The membranes were blocked at room temperature with 5% non-fat milk for 1 h, and incubated with primary antibodies against MVK (1:1,000; cat. no. 12228-1-AP; ProteinTech Group, Inc.), cytokeratin 1 (1:1,000; cat. no. 16848-1-AP; ProteinTech Group, Inc.), involucrin (1:1,000; cat. no. 55328-1-AP; ProteinTech Group, Inc.), lamin A (1:1,000; cat. no. 10298-1-AP; ProteinTech Group, Inc.), HRAS (1:1,000; cat. no. 18295-1-AP; ProteinTech Group, Inc.), Rho E (1:1,000; cat. no. D223036; Sangon Biotech Co., Ltd.), Rho B (1:1,000; cat. no. 14326-1-AP; ProteinTech Group, Inc.), Rho A (1:1,000; cat. no. 10749-1-AP; ProteinTech Group, Inc.), RAC1 (1:1,000; cat. no. 24072-1-AP; ProteinTech Group, Inc.) and CDC42 (1:1,000; cat. no. 10155-1-AP; ProteinTech Group, Inc.) at 4 ̊C overnight, respectively.

Techniques: Expressing, Over Expression, Quantitative RT-PCR, Western Blot, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Comprehensive Compositional Analysis of the Slit Lamp Bacteriota

doi: 10.3389/fcimb.2021.745653

Figure Lengend Snippet: Bar chart of relative abundances of ASVs classified on genus level. Taxa with relative abundance of less than 1% were collectively summarized as ‘Other’. (A) Taxonomic composition of the slit lamp bacteriota from TC1 (Tertiary Center) and TC2 respectively and the respective sample sites (contact area – TC1: n = 28; ocular – TC1: n = 28; contact area - TC2: n = 14; ocular – TC2: n = 13). (B) Composition of mock taxa and controls (expected = expected mock abundances; mock = mock standard using Bact-0341f/Bact-0785r primers; blank = blank negative control).

Article Snippet: The mock community consisted of 6 typical skin bacterial species in equal total cell abundances ( Acinetobacter johnstonii ATCC 17090, Corynebacterium striatum ATCC 6940, Micrococcus luteus ATCC 4698, Cutibacterium acnes ATCC 11828, Staphylococcus epidermidis ATCC 12228 and Streptococcus mitis ATCC 49456, 16.7% each).

Techniques: Negative Control