Determination of amino acids in saliva using capillary electrophoresis with fluorimetric detection

In the present study a sensitive method for the quantification of main free amino acids in saliva using capillary electrophoresis with laser induced fluorescence detection was developed. As background electrolyte 20 mM borate buffer pH 9.5 was used. Amino acids were separated after derivatization with fluorescein isothiocyanate (FITC) and the conditions for derivatization were optimized. The main amino acids occurring in saliva (Pro, Ser, Gly and Glu) were separated in less than 7 min. The parameters of validation such as linearity of response, precision and detection limits were determined. The detection limits were obtained in the range from 0.1 to 2.4 nM. The developed method was employed for determination of amino acids in real saliva samples.     

The effect of skinfold on frequency of human muscle mechanomyogram.

The aim of the study was to assess the effect of skinfold thickness on median and peak frequency of mechanomyographic (MMG) signal in relation to subject's age, gender and force during voluntary contraction of elbow flexor and extensor muscles. Seventy-nine healthy subjects participated in the study: 22 young females (age 20.1+/-1.1 years), 22 young males (age 23.4+/-1.1 years), 17 elderly females (age 64.9+/-5.1 years), and 18 elderly males (age 67.4+/-6.2 years). Three identical MMG probes were used to record MMG signals from above the triceps brachii (TB), biceps brachii (BB), and brachioradialis (BR) muscles simultaneously with the force signal. The results showed that the tissue between the muscle and the skin surface has a major contribution to the median and a minor contribution to the peak MMG frequencies independent of subjects' age (with force having more than 2 folds a smaller effect). During antagonistic function of the main elbow flexors and extensors, there is a decreasing effect of skinfold thickness and an increasing effect of force on the MMG frequency, and the relative contribution of both factors to the MMG signal is age related, especially in the TB and BR muscles. The BR muscle differs from the TB and BB muscles in regard to the effects of skinfold thickness and force on the MMG frequency, as well as in the effect of age on the relationship between the MMG frequency and skinfold thickness and force. The effect of age on the relative contribution of skinfolds and force to MMG frequency is specific for muscle and its function. It was concluded that studies that report MMG frequency with different values of skinfold thickness cannot be easily compared, especially when maximally activated prime movers are tested. A use of force and skinfold thickness as covariates is recommended when an MMG frequency is analyzed in subjects differing in the skinfold thickness.     

Are Homo sapiens nonsupranuchal fossa and neanderthal suprainiac fossa convergent traits?

The autapomorphic status of the Neanderthal suprainiac fossa was recently confirmed. This was a result of a detailed analysis of the internal bone composition in the area of the suprainiac depression on Neanderthal and Homo sapiens specimens. However, while anatomical differences between Neanderthal suprainiac fossa and the depression in the inion region of the occipital bone of fossil and recent Homo sapiens have been discussed in detail, the etiology of these structures has not been resolved. In this article, the hypothesis that the Homo sapiens non-supranuchal fossa and the Neanderthal suprainiac fossa both formed to maintain the optimal shape of the occipital plane (to minimize strain on the posterior cranial vault) is tested. First, the variation in the expression of the fossa above inion in the crania of recent Homo sapiens from European, African, and Australian samples was examined, and the degree of structural similarity between these depressions and the Neanderthal suprainiac fossa was assessed. Next, the relationship between the shape of the occipital squama in the midsagittal plane and two particular features (the degree of the occipital torus development and the occurrence of a depression in the inion region that is not the supranuchal fossa) were analyzed. Based on the results, it is suggested that the Homo sapiens non-supranuchal fossa and Neanderthal suprainiac fossa are convergent traits.     

'happy on norflurazon' (hon) mutations implicate perturbance of plastid homeostasis with activating stress acclimatization and changing nuclear gene expression in norflurazon-treated seedlings

Various mutant screens have been undertaken to identify constituents involved in the transmission of signals from the plastid to the nucleus. Many of these screens have been performed using carotenoid-deficient plants grown in the presence of norflurazon (NF), an inhibitor of phytoene desaturase. NF-treated plants are bleached and suppress the expression of nuclear genes encoding chloroplast proteins. Several genomes uncoupled (gun) mutants have been isolated that de-repress the expression of these nuclear genes. In the present study, a genetic screen has been established that circumvents severe photo-oxidative stress in NF-treated plants. Under these modified screening conditions, happy on norflurazon (hon) mutants have been identified that, like gun mutants, de-repress expression of the Lhcb gene, encoding a light-harvesting chlorophyll protein, but, in contrast to wild-type and gun mutants, are green in the presence of NF. hon mutations disturb plastid protein homeostasis, thereby activating plastid signaling and inducing stress acclimatization. Rather than defining constituents of a retrograde signaling pathway specifically associated with the NF-induced suppression of nuclear gene expression, as proposed for gun, hon mutations affect Lhcb expression more indirectly prior to initiation of plastid signaling in NF-treated seedlings. They pre-condition seedlings by inducing stress acclimatization, thereby attenuating the impact of a subsequent NF treatment.     

Effect of rhIL-15 on TLR2 expression and apoptosis of PMN from patients with Lyme disease

In this study we estimated the expression of TLR2 and apoptosis of PMN in patients with Lyme disease. The cells were isolated from heparinized whole blood by Gradisol G gradient and incubated 18 h with rhIL-15 and LPS. Expression of TLR2 was estimated in lysates of PMN by western blot, apoptosis of PMN by immunofluorescent analysis. The results obtained revealed the higher expression of TLR2 in PMN and higher percentage of apoptosis PMN in patients with Lyme disease compared with control. We observed an effect of rhIL-15 on the increased expression of TLR2 in PMN and the increased survival of PMN isolated from patients with Lyme disease. These findings suggest that IL-15 has the ability to modulate of neutrophil response against Borrelia burgdorferi.     

VEGF, IL-18 and NO production by neutrophils and their serum levels in patients with oral cavity cancer

It is known that the relationship between pro-angiogenic and anti-angiogenic factors is responsible for the presence and intensity of neoangiogenesis. The angiogenic factors are produced by tumour cells and/or by tumour-infiltrating inflammatory cells such as macrophages or polymorphonuclear leukocytes (PMN). In the present study we compared VEGF secretion with IL-18 and NO release by PMN derived from oral cavity cancer patients. Knowledge of the relationship between mediators above could help in better understanding the role of PMN in angiogenesis in this patient group. The results from culture supernatants of PMN were confronted with the serum levels of parameters examined. We found an interesting relationship between VEGF and IL-18 concentrations in the culture supernatants of PMN derived from patients with oral cavity cancer. High production of VEGF was associated with low production of IL-18 by PMN derived from patients before treatment. During examinations after treatment we found lower concentrations of VEGF and higher concentrations of IL-18 than those in the study before treatment. In contrast to VEGF and IL-18, the NO production by PMN of cancer patients, before and after treatment, was unchanged. We also demonstrated markedly elevated serum levels of VEGF as well as IL-18 according to the progression of the disease. Results obtained indicate that relations between VEGF and IL-18 released by PMN may promote neoangiogenesis and may be important for benign tumour cells to acquire metastatic phenotype in the early stage of oral cavity cancer. Furthermore, our results suggest that the concentrations of VEGF and IL-18 in the serum are sensitive tumour markers in this patient group before and after treatment.     

CAG repeat polymorphism in the androgen receptor (AR) gene of SBMA patients and a control group

Spinobulbar muscular atrophy (SBMA) is an X-linked form of motor neuron disease characterized by progressive atrophy of the muscles, dysphagia, dysarthria and mild androgen insensitivity. SBMA is caused by CAG repeat expansion in the androgen receptor gene. CAG repeat polymorphism was analysed in a Polish control group (n = 150) and patients suspected of SBMA (n = 60). Normal and abnormal ranges of CAG repeats were established in the control group and in 21 patients whose clinical diagnosis of SBMA was molecularly confirmed. The ranges are similar to those reported for other populations.     

Annealing of normal and mutant wheat starches. LM, SEM, DSC, and SAXS studies

Structure and thermodynamic properties of native and annealed wheat starches with different amylose content (from 1.5% to 39.5%) have been studied by high-sensitivity differential scanning microcalorimetry (HSDSC), small-angle X-ray diffraction (SAXS), light (LM), and scanning electron microscopy (SEM). Starch morphology, the values of the melting cooperative unit, the thickness of crystalline lamellae and the size of amylopectin clusters as well as thermodynamic parameters characterizing surface of the face side in starch crystals were determined. Some suppositions based on different physical approaches are used for a discussion of the results concerning structural reorganization of starches on different levels of macromolecular organization.