Preliminary phenotypic characterization of newly isolated halophilic microorganisms by footprinting: a rapid metabolome analysis

The emerging need for rapid screening and identification methods for microbiological purposes necessitates the combined uses of high-tech instruments. In this work, electrospray ionization mass spectrometry was used to visualize the relation of ten newly isolated moderately halophilic microorganisms, to Halomonas salina DSMZ 5928 and Halomonas halophila DSMZ 4770. The method was based on the global analysis of the metabolites in culture media and is termed as metabolic footprinting. Since it was not possible to gain insight into the similarities solely based on the visual inspection of the chromatograms, principal component (PC) analysis was applied on the data. Three PCs alone were able to explain 99% of the information in the data set. The score plots revealed the relation of the new isolates to the two type strains whereas the loading plots gave important clues on the significant ions responsible for the observed clustering. Loading plots also indicated inversely correlated ions that give clues on differing metabolic pathways. The work described here offers a potentially useful way for preliminary rapid phenotypic characterization of new and closely related isolates and a method for screening of similar microorganisms for different and valuable secondary metabolites.     

Alkaline phosphatase interference in immuno-enzymatic assays

BackgroundAlkaline phosphatase (ALP) enzymes are widely used as signal amplifiers in immunoenzymatic methods. Conditions that cause ALP elevations, such as bone or liver diseases, can cause interference in immunoenzymatic methods. We aimed to examine ALP''s effect on immunoenzymatic assay by adding isolated pure ALP to the prepared serum pool.MethodsWe prepared a serum pool and divided it into 4 groups. By adding isolated pure ALP at different concentrations to each group, we obtained sample groups containing ALP enzyme at concentrations of 85 U/L, 340 U/L, 870 U/L, and 1570 U/L. 20-repetition of bhCG, ferritin, FT4, TSH, troponin I, and Vit B12 tests were performed in each group. The coefficient of variation, bias, and total error was calculated. All groups were compared by using the Friedman test for paired samples.ResultsAfter ALP addition, the calculated total error values of FT4, bhCG and troponin I tests were above the acceptable error limits. There were statistically significant differences in bhCG, FT4, troponin I, and Vit B12 tests compared to the baseline ALP level (P<0.0125).ConclusionsIsolated ALP elevations can be a source of interference for immunoenzymatic methods.     

Increasing the Bioaccessibility of Antioxidants in Tomato Pomace Using Excipient Emulsions

Plant-based foods contain numerous bioactive constituents (“nutraceuticals”) that have beneficial effects on human health. However, their oral bioavailability is often relatively low, which limits their potential efficacy. The bioavailability of nutraceuticals can be increased through the utilization of excipient foods whose compositions and structures are specifically designed to increase the amount of nutraceuticals absorbed in an active form. In this study, olive oil excipient emulsions were designed to increase the bioaccessibility of lycopene and other natural antioxidants in tomato pomace. These emulsions consisted of 8 wt% olive oil and 1 wt% Tween 20 or Tween 80 and were prepared using a microfluidizer operated under different processing conditions (12,000 or 20,000 psi; 3 or 5 passes). Changes in particle size, charge, and bioaccessibility were assessed when tomato pomace-emulsion mixtures were exposed to simulated gastrointestinal digestion. The mean particle diameter of the particles in the excipient emulsions increased after digestion (416 to 446 nm) compared to the values before digestion (200 to 220 nm). The presence of excipient emulsions significantly increased the bioaccessibility of lycopene in tomato pomace compared to oil-free control samples. For instance, lycopene bioaccessibility was > 82% when the tomato pomace was mixed with excipient emulsions but only 29% when it was mixed with oil-free buffer solutions. The presence of excipient emulsions also increased the total phenolic content of the tomato pomace. For instance, the phenolic content was considerably higher in the presence of excipient emulsions (1489 to 2055 mg GAE /100 g FW) than in their absence (939 mg GAE /100 g FW). However, the excipient emulsions did not increase naringenin bioaccessibility, which was attributed to the fact that it was not strongly hydrophobic. The efficacy of the excipient emulsions was only modestly dependent on emulsifier type and homogenization conditions. In conclusion, excipient emulsions can be designed to enhance the bioaccessibility of strongly hydrophobic nutraceuticals in tomato-based products, which may boost their healthiness.

Graphical abstract

     

Simultaneous immunofluorescence and histology in pemphigus vulgaris using ex vivo confocal laser scanning microscopy

Ex vivo confocal laser scanning microscopy (ex vivo CLSM) provides rapid, high-resolution imaging and immunofluorescence examinations of the excised tissues. We aimed to evaluate the applicability of ex vivo CLSM in histomorphological and direct immunofluorescence (DIF) examination of pemphigus vulgaris (PV). 20 PV sections were stained with fluorescent-labeled anti-IgG and anti-C3 using various dilutions and incubation periods. Subsequently, the determined ideal staining protocol was applied on 20 additional PV and 20 control sections. Ex vivo CLSM identified intraepidermal blisters and acantholytic cells in 80% and 60% of PV patients, respectively. The sensitivity of ex vivo CLSM in detecting intraepidermal fluorescence was 90% both with IgG and C3. The specificity of staining for IgG and C3 was 70% and 90%, respectively. Histomorphological and immunofluorescence features of PV could be detected within the same ex vivo CSLM session showing a comparable performance to conventional histopathology and DIF microscopy.     

Trace Element Levels and Oxidant/Antioxidant Status in Patients with Alcohol Abuse

Biological Trace Element Research - Alcohol abuse is a well-known cause of imbalance in trace element levels and oxidant/antioxidant status of individuals with long time consumption. However, the...     

Circulating nuclear factor-kappa B mediates cancer-associated inflammation in human breast and colon cancer

BackgroundInflammation is recognized as a hallmark feature of cancer development and progression. The aim of our study was to investigate the significance of serum nuclear factor kappa-B (NF-κB) levels as a circulating marker in the monitoring of inflammation in breast and colon cancer; to show the relationship between NF-κB with inflammatory parameters as tumour necrosis factor-α (TNF-α), soluble TNF-related apoptosis-inducing ligand (sTRAIL), interleukin-6 (IL-6), pentraxin-3 (PTX-3), procalcitonin (PCT), and C-reactive protein (CRP) levels.MethodsSerum NF-κB, TNF-α, sTRAIL, IL-6, PTX-3, PCT, and serum CRP levels were measured using enzyme-linked immunosorbent assay (ELISA) in 40 patients with breast cancer, 40 patients with colon cancer and 30 healthy controls.ResultsThe serum NF-κB, TNF-α, IL-6, PTX-3, PCT, and serum CRP concentration was significantly higher, and the serum sTRAIL concentration was significantly lower in the patients with breast and colon cancer than in healthy controls. NF-κB was positively correlated with CRP and negatively correlated with sTRAIL.ConclusionsThese results suggest that increased NF-κB may decrease the clinical efficacy of sTRAIL in solid tumour cells. There is a relationship between inflammation and carcinogenesis so that the development of cancer occurs with chronic inflammation in breast and colon. The study results have shown that colon and breast cancer patients have increased systemic inflammation, as measured by increased circulating cytokines, and acute-phase proteins, or by abnormalities in circulating cells. NF-κB may combine with other markers of the systemic inflammatory response in prognostic scores in cancer. In addition to surgical resection of the tumour, and conventional radio and chemotherapy for cancer treatment, the use of sTRAIL or other agonists for cancer therapy appeared a new potential therapy.     

Investigation of the relationship between kinesiophobia and sensory processing in fibromyalgia patients

Abstract

Purpose/aim: The aims of this study are to investigate the relationship between kinesiophobia and sensory processing in fibromyalgia (FM) patients and obtain new information about kinesiophobia in light of these results.

Materials and methods: This study has been conducted with 82 literate subjects aged 18–65?years and diagnosed with FM based on ACR 2010 diagnostic criteria. Fibromyalgia Impact Questionnaire was used for measuring functional status in FM patients, The Tampa Scale of Kinesiophobia was used for determine the levels of kinesiophobia and Adolescent/Adult Sensory Profile was used for determine the characteristics of sensory process patterns of the individuals.

Results: Among the participants, sensory sensitivity scores of 65.85% and sensation avoiding scores of 40.24% were higher than that in the general population. Sensation seeking scores of 48.78% the subjects were lower compared to the general population. A significant and weak positive correlation was found between the kinesiophobia scores and responses of sensory sensitivity and sensation avoiding (r?=?0.23, p?=?0.04; z?=?0.29, p?=?0.01)

Conclusion: This is the first study conducted to investigate the relationship between kinesiophobia and sensory processing in fibromyalgia patients. However, different studies investigating this subject are warranted in order to be able to generalize the findings and increase the value of evidence.