Truncated Gaussian-Bessel beams for short-pulse processing of small-aspect-ratio micro-channels in dielectrics.

In order to control the length of micro-channels ablated at the surface of dielectrics, we use annular filtering apertures for tailoring the depth of focus of micrometric Gaussian-Bessel beams. We identify experimentally and numerically the appropriate beam truncation that promotes a smooth axial distribution of intensity with a small elongation, suitable for processing micro-channels of small aspect ratio. Single-shot channel fabrication is demonstrated on the front surface of a fused silica sample, with sub-micron diameter, high-quality opening, and depth of few micrometers, using 1 ps low-energy (< 0.45 µJ) pulse. Finally, we realize 10 × 10 matrices of densely packed channels with aspect ratio ~5 and a spatial period down to 1.5 µm, as a prospective demonstration of direct laser fabrication of 2D photonic-crystal structures.

Isomorphic coalescence of aster cores formed in vitro from microtubules and kinesin motors.

We report fluorescence microscopy studies of the formation of aster-like structures emerging from a cellular element-based active system and a novel analysis of the aster condensation. The system consists of rhodamine labeled microtubules which are dynamically coupled by functionalized kinesin motor proteins cross-linked via streptavidin-coated quantum dots (QDs). The aster-shaped objects contain core structures. The cores are aggregates of the QD-motor protein complexes, and result from the dynamic condensation of sub-clusters that are connected to each other randomly. The structural specificity of the aster core reflects a configuration of the initial connectivity between sub-clusters. Detailed image analysis allows us to extract a novel correlation between the condensation speed and the sub-cluster separation. The size of the core is scaled down during the condensation process, following a power law dependence on the distance between sub-clusters. The exponent of the power law is close to two, as expected from a geometric model. This single exponent common to all the contractile lines implies that there exists a time regime during which an isomorphic contraction of the aster core continues during the condensation process. We analyze the observed contraction by using a model system with potential applicability in a wide range of emergent phenomena in randomly coupled active networks, which are prevalent in the cellular environment.

Dihalide and pseudohalide radical anions as oxidizing agents in nonaqueous solvents.

The dihalide and pseudohalide radical anions, strong one-electron oxidants, can be selectively generated in aqueous solutions by pulse radiolysis. Radiolysis of salts of the bulky 1-butyl-3-methylimidazolium cation with Cl(-), Br(-), SCN(-), and N(3)(-) anions allows efficient generation of the same species (Cl(2)(*-), Br(2)(*-), (SCN)(2)(*-), and N(6)(*-) radical anions) also in neat ionic liquids and in nonaqueous solvents with the addition of small amounts of the salt. The oxidative power of dichloride radical anion, lowered in the dichloromethane as compared to the aqueous solution, remains sufficient for oxidation of N,N,N',N'-tetramethyl-p-phenylenediamine (TMPD, (2)k = (2.0 +/- 0.1) x 10(9) M(-1) s(-1)), phenothiazine (PZ, (2)k = (2.1 +/- 0.1) x 10(9) M(-1) s(-1)), 10-methylphenothiazine (MPZ, (2)k = (9.3 +/- 0.1) x 10(7) M(-1) s(-1)), and 3-methylindole (scatole, SCT, (2)k = (6.8 +/- 0.1) x 10(7) M(-1) s(-1)). It diminishes on going to Br(2)(*-) (reacts only with TMPD, (2)k = (5.4 +/- 0.3) x 10(8) M(-1) s(-1), and PZ, (2)k = (4.8 +/- 0.3) x 10(8) M(-1) s(-1)) while (SCN)(2)(*-) and N(6)(*-) radical anions oxidize only TMPD, (2)k = (5.1 +/- 0.5) x 10(8) M(-1) s(-1) and (2)k approximately 10(8) M(-1) s(-1), respectively.

CD8 infiltration is associated with disease control and tobacco exposure in intermediate-risk oropharyngeal cancer.

Oropharyngeal squamous cell carcinoma (OPSCC) incidence is increasing at a nearly epidemic rate, largely driven by the human papillomavirus (HPV). Despite the generally favorable clinical outcomes of patients with HPV driven (HPV+) OPSCC, a significant subset of HPV tumors associated with tobacco exposure have diminished treatment response and worse survival. The tumor immune microenvironment (TIME) has been shown to be a critical driver of treatment response and oncologic outcomes in OPSCC generally and HPV+ OPSCC more specifically. However, the impact of tobacco exposure on the TIME in OPSCC patients remains unclear. We analyzed the relationship between TIME, tobacco exposure and clinical outcomes in OPSCC patients (n = 143) with extensive tobacco exposure (median pack-years = 40). P16 overexpression, a surrogate marker of HPV association, was a strong predictor of relapse-free (RFS) and overall survival (OS) (p < 0.001, p < 0.001 respectively) regardless of tobacco exposure and associated strongly with differential infiltration of the tumor by both CD3 and CD8 lymphocytes measured via immunohistochemistry (p < 001, p < 0.001 respectively). CD3 and CD8 infiltration was a strong predictor of RFS and OS and associated strongly with disease stage (AJCC 8th Edition Staging Manual). Tobacco exposure correlated significantly (p < 0.001) with decreased CD8 infiltration in p16+ OPSCC tumors. Our findings demonstrate that the HPV+ OPSCC clinical outcomes are strongly correlated with the TIME, which is potentially modulated by tobacco exposure. Immunomodulatory strategies targeting this disease in smokers must take into consideration the potential modifying effects of tobacco exposure on treatment effectiveness and clinical outcomes.

Clinical evaluation of proinflammatory cytokine inhibitors (sTNFR I, sTNFR II, IL-1 ra), anti-inflammatory cytokines (IL-10, IL-13) and activation of neutrophils after burn-induced inflammation.

The study was aimed at evaluating the involvement of sTNFR I, sTNFR II, IL-1 ra, IL-10, IL-13 and reactive oxygen species (ROS) in systemic inflammatory response syndrome (SIRS) development in severely burned children and at assessing the prognostic value of the immunological markers studied. The study comprised 37 patients (17 burned children and 20 controls). Serum levels of the markers determined by means of ELISA and respiratory burst of neutrophils as well as p55 and p75 tumour necrosis factor-alpha (TNF-alpha) receptor expression using flow cytometry were evaluated twice. The burned children presented significantly higher levels of IL-10 and cytokine inhibitors within the first 6-24 h after injury compared with controls (P < 0.05). The decreased oxygen metabolism of neutrophils and increased TNF-alpha receptor expression were found on admission. Moreover, a significant decrease in initially high sTNFR I, sTNFR II, IL-1 ra, IL-10, IL-13 concentrations (P < 0.05) and reduced expression of TNF-alpha receptors (P < 0.05) were observed after burn therapy, whereas ROS generation evidently augmented (P < 0.05). Four of our children who developed hypovolaemic shock revealed a significantly lower ROS generation and higher concentrations of soluble TNF-alpha receptors and IL-1 ra together with IL-10, IL-13 compared with children with good outcome (P < 0.05). Our results revealed the involvement of both ROS, soluble TNF-alpha receptors and IL-1 ra in the development of SIRS in burned children; their monitoring allows for an assessment of the systemic inflammatory reaction activity. The neutrophil BURSTTEST and IL-1 ra might have been clinically helpful markers of SIRS prognosis.

Heat shock response in gastrointestinal tract.

Heat shock response is one of the defense mechanisms common to eukaryotic and prokaryotic cells. The highly conserved and ubiquitous heat shock proteins (HSPs) are essential for cell survival during stress. Stress tolerance, i.e., adaptation of cells to stress conditions, is a characteristic feature of heat shock response. The lumen of the gastrointestinal tract is an external environment to the body. Epithelium of the digestive tract is exposed to various stress factors inducing the heat shock response, e.g., bacteria and their toxins, food borne chemical compounds, drugs and diet deficiencies. Other factors like plant lecitins, glutamine or short fatty acids are mild stressors and can modulate the heat shock response in cells. All these factors are presumed to influence the normal microflora that is an integral part of the digestive tract. This review is focused on the induction/modulation of heat shock proteins expression in the epithelium of the gastrointestinal tract by various factors, on the protective role of HSPs and mechanisms leading to stress protection inside the gut. Heat shock response is one of the key mechanisms of maintenance of gastrointestinal tract homeostasis. It is involved in pathogenic bacteria adaptation to life in the digestive tract, especially in colony formation and in their role in infectious processes.

Experimental vaccine induces Th1-driven immune responses and resistance to Neisseria gonorrhoeae infection in a murine model.

Female mice were immunized intravaginally with gonococcal outer membrane vesicles (OMVs) plus microencapsulated interleukin-12 (IL-12), and challenged using an established model of genital infection with Neisseria gonorrhoeae. Whereas sham-immunized and control animals cleared the infection in 10-13 days, those immunized with OMV plus IL-12 cleared infection with homologous gonococcal strains in 6-9 days. Significant protection was also seen after challenge with antigenically distinct strains of N. gonorrhoeae, and protective anamnestic immunity persisted for at least 6 months after immunization. Serum and vaginal immunoglobulin G (IgG) and IgA antibodies were generated against antigens expressed by homologous and heterologous strains. Iliac lymph node CD4+ T cells secreted interferon-γ (IFNγ), but not IL-4, in response to immunization, and produced IL-17 in response to challenge regardless of immunization. Antigens recognized by immunized mouse serum included several shared between gonococcal strains, including two identified by immunoproteomics approaches as elongation factor-Tu (EF-Tu) and PotF3. Experiments with immunodeficient mice showed that protective immunity depended upon IFNγ and B cells, presumably to generate antibodies. The results demonstrated that immunity to gonococcal infection can be induced by immunization with a nonliving gonococcal antigen, and suggest that efforts to develop a human vaccine should focus on strategies to generate type 1 T helper cell (Th1)-driven immune responses in the genital tract.

Evaluation of the Escherichia coli HK82 and BS87 strains as tools for AlkB studies.

Within a decade the family of AlkB dioxygenases has been extensively studied as a one-protein DNA/RNA repair system in Escherichia coli but also as a group of proteins of much wider functions in eukaryotes. Two strains, HK82 and BS87, are the most commonly used E. coli strains for the alkB gene mutations. The aim of this study was to assess the usefulness of these alkB mutants in different aspects of research on AlkB dioxygenases that function not only in alkylated DNA repair but also in other metabolic processes in cells. Using of HK82 and BS87 strains, we found the following differences among these alkB(-) derivatives: (i) HK82 has shown more than 10-fold higher MMS-induced mutagenesis in comparison to BS87; (ii) different specificity of Arg(+) revertants; (iii) increased induction of SOS and Ada responses in HK82; (iv) the genome of HK82, in comparison to AB1157 and BS87, contains additional mutations: nalA, sbcC, and nuoC. We hypothesize that in HK82 these mutations, together with the non-functional AlkB protein, may result in much higher contents of ssDNA, thus higher in comparison to BS87 MMS-induced mutagenesis. In the light of our findings, we strongly recommend using BS87 strain in AlkB research as HK82, bearing several additional mutations in its genome, is not an exact derivative of the AB1157 strain, and shows additional features that may disturb proper interpretation of obtained results.

Thermoresponsive polymers as promising new materials for local radiotherapy.

We describe a novel thermoresponsive polymeric drug delivery system based on poly(N-isopropylacrylamide) with isotopically labellable end groups [l-tyrosinamide or diethylenetriaminepentaacetic acid (DTPA)] designed for local radiotherapy. The polymers are readily soluble in isotonic aqueous sodium chloride at room temperature and the phase separation is complete at body temperature as proved by DSC measurements. Sufficent binding capacity for radionuclides and chemical stability are demonstrated on 125I and 90Y-labelled polymers.

Popular species of edible mushrooms as a good source of zinc to be released to artificial digestive juices.

Because fruiting bodies of edible mushrooms accumulate elements very effectively, in this study for the first time we aimed at determining the degree of the release of zinc(II) ions to artificial digestive juices imitating the human gastrointestinal tract from freeze-dried popular edible mushroom fruiting bodies, such as Agaricus bisporus, Boletus badius and Cantharellus cibarius. For the analysis, anodic stripping voltammetry method was used. The amount of zinc released to artificial saliva within 1 minute ranged from 0.03 to 1.14 mg/100 g d.w. In gastric juice, the amounts were higher and ranged from 0.75 to 2.07 mg/100 g d.w. depending on the incubation time. After incubation of the freeze-dried edible mushroom fruiting bodies for 1 minute in artificial saliva, 15 in artificial gastric juice and then 150 minutes in artificial intestinal juice, it was found that the concentration of the released zinc in artificial intestinal juice was the highest and amounted to 6.44 mg/100 g d.w. The total average amount of zinc released from Boletus badius was the highest and this was estimated at 4.13 mg/100 g d.w. For the remaining two investigated species of A. bisporus and C. cibarius, the total amounts of zinc released into artificial digestive juices were only slightly lower and were estimated at 2.23 and 3.29 mg/100 g d.w. on average, respectively. It was demonstrated for the first time that mushrooms release zinc to artificial digestive juices imitating conditions in the human digestive tract and are a good source of this element.

Phenotype and function of nasal dendritic cells.

Intranasal (i.n.) vaccination generates immunity across local, regional, and distant sites. However, nasal dendritic cells (DCs), pivotal for the induction of i.n. vaccine-induced immune responses, have not been studied in detail. Here, by using a variety of parameters, we define nasal DCs in mice and humans. Distinct subsets of "classical" DCs, dependent on the transcription factor zbtb46 were identified in the murine nose. The murine nasal DCs were Fms-related tyrosine 3 kinase ligand responsive and displayed unique phenotypic and functional characteristics, including the ability to present antigen, induce an allogeneic T-cell response, and migrate in response to lipopolysaccharide or live bacterial pathogens. Importantly, in a cohort of human volunteers, BDCA-1(+) DCs were observed to be the dominant nasal DC population at steady state. During chronic inflammation, the frequency of both BDCA-1(+) and BDCA-3(hi) DCs was reduced in the nasal tissue, associating the loss of these immune sentinels with chronic nasal inflammation. The present study is the first detailed description of the phenotypic, ontogenetic, and functional properties of nasal DCs, and will inform the design of preventative immunization strategies as well as therapeutic modalities against chronic rhinosinusitis.

Exposure of Escherichia coli to intestinal myoelectrical activity-related electric field induces resistance against subsequent UV(254 nm) (UVC) irradiation.

Survival of Escherichia coli K-12 AB1157 irradiated with UVC (UV(254 nm)) was enhanced after pre-treatment with a low-tension electric field (EF). The EF used was identical to the electrical field generated by the small intestine (myoelectrical migrating complex--MMC), registered in a healthy calf and transmitted into the memory of an EF generator. The EF emitted by the generator was transmitted via electrodes placed in shaken bacterial cultures. The protective effects of the EF on the E. coli survival after exposure to UV were: (i) observed only for the dnaJ(+)dnaK(+) strain, and not for the DeltadnaJdnaK heat shock mutant; (ii) strictly dependent on the temperature at which the bacteria were grown; (iii) most obvious when the bacteria were incubated at 37 degrees C. Moreover, the MMC-related EF and a higher temperature (40 degrees C) show a similar protective effect against UV-irradiation. The results point to the involvement of the heat shock response in the low-tension EF-induced protection of bacterial cells against UVC-irradiation. Additionally, treatment with the MMC-related EF affects total protein contents and their pattern in E. coli cells. The EF-treatment did not show any influence on the level of the argE3(ochre) --> Arg(+) reversions.

The development of the blood-brain barrier in alcohol-exposed rats.

Circulating horseradish peroxidase (HRP) was used as a tracer to determine if the blood-brain barrier to protein was altered by dietary prenatal alcohol exposure. Animals were prepared for light microscopic visualization of HRP after HRP infusion on gestational days 16, 18, 20, 22 and postnatal day 4. There was no consistent evidence of HRP leakage through the BBB in the alcohol-exposed animals compared to control animals. Capillary endothelial cells and perivascular astrocytic endfeet were morphologically characterized by electron microscopy in rat optic nerve and cerebellum following dietary prenatal and postnatal ethanol exposure. Photomontages of optic nerve capillaries from G20 and P5 animals and cerebellar capillaries from P15 animals were examined for evidences of effects of alcohol on the development of the capillaries and adjacent astroglial endfeet. There was no consistent evidence of any alcohol-induced effect that could indicate a disruption of the vessel, the endothelial tight junctions, the perivascular glial limiting membranes, or the extent of vascular ensheathment by astrocytic endfeet.

Mercury in water and sediments of the southern Baltic sea.

Total mercury concentrations [HgT] were measured in samples of water and sediments collected from the southern Baltic Sea from the period 1993-1995. Picomolar concentrations of HgT were found in offshore surface water. Higher levels were measured in samples from the plum of the Vistula. An exceptionally high concentration of mercury (25 pM HgT, 5 pM methylmercury) was measured in the near bottom, quasi-anoxic water of the Gdañsk Deep and explained by the release of mercury from fine grained sediments covering the bottom. In the sediments, the largest contents of HgT were found in fine grained, organic rich deposits of the Baltic deeps. The increase in mercury contents in the upper layers of sediments deposited within the last 100-150 years was attributed to the anthropogenic load of mercury and its increased scavenging to sediments with organic matter.

Speciation of heavy metals in marine sediments vs their bioaccumulation by mussels.

Total contents and speciation of selected heavy metals, including Al, Fe, Co, Ni, Pb, Zn, Cu, Cr, were measured in sediment samples and mussels Mya arenaria and Astarte borealis collected in the Horsund Fjord off Spitsbergen (Norwegian Sea) and the Bay of Gdansk (Baltic Sea). The investigation aimed at revealing differences in the accumulation pattern of heavy metals in mussels inhabiting sediments characterized by varying metal bioavailability. The contents of metals adsorbed to sediments and associated with iron and manganese hydroxides, which were obtained by sequential extraction, were utilized as a measure of metal bioavailability. The contents of Cd, Pb, Zn, Cu and Cr in mussels collected off Spitsbergen were generally lower than those in mussels from the Baltic Sea. In sediments collected off Spitsbergen the bioavailable fraction represented a small proportion (0-3.7% adsorbed metals and 0-11% associated with metals hydroxides) of total heavy metal contents. In sediments from the Baltic Sea the percentages of metals adsorbed and bound to hydroxides were 1-46% and 1-13%, respectively. The differences in bioavailable metal contents measured in sediments were utilized to explain the different contents of metals in mussels collected in the corresponding sites.

Content of folic acid and free homocysteine in blood serum of human papillomavirus-infected women with cervical dysplasia.

The authors estimated the concentrations of folic acid and free homocysteine in the blood serum of women with CIN III (cervical intraepithelial neoplasia-Burghard's classification) infected with DNA HPV (human papillomaviruses) of type 16 and/or 18. The control group consisted of 49 patients with normal cytological smears without HPV infection. Types 16 and/or 18 DNA HPV were found in 50 patients. This women qualified for the studied group. The sequence of DNA HPV type 16 and/or 18 was identified with the PCR method (polymerase chain reaction). The high-performance liquid chromatography (HPLC) method was employed to evaluate the levels of folic acid and free homocysteine in the blood serum of the examined patients. Significantly lower levels of folic acid and higher levels of free homocysteine were observed in the blood serum of HPV-positive patients with CIN III. The correlation was found between serum concentrations of folic acid and free homocysteine in both groups.

Clinical evaluation of patients with idiopathic juvenile osteoporosis.

The aim of this study is to evaluate the occurrence of idiopathic juvenile osteoporosis by clinical, radiographic and biochemical methods in 7 patients aged 10 years to 18 years. The prevailing clinical symptoms included pain in the lower limbs and spine, gait disturbances and weak muscles; confirmed fractures were seen in four out of seven patients. In all patients, bone mineral density was determined by dual-energy X-ray absorptiometry and calcium-phosphorus metabolism assays were performed. Clinical evaluation of our patients revealed a wide variety of classic clinical symptoms, and also radiographic and biochemical features. In this complex evaluation, we can distinguish two types of disease occurrence, mild and acute.

The bioavailability of injectable, amorphous form of aztreonam.

Amorphos injectable form of aztreonam (BIOKTAM) was prepared. It was shown that after intramuscular or intravenous administration there are not any considerable differences in the bioavailability of aztreonam from BIOKTAM and of the drug from AZACTAM.

[Assessment of early psychic dysfunctions due to metallic mercury]. / Wykrywanie wczesnych zaburzen funkcji psychicznych wywolanych dzialaniem rteci metalicznej.

The aim of the study was to find the most sensitive psychological test battery, useful for evaluation of the earliest dysfunctions in workers exposed to metallic mercury (Hg). To assess the above mentioned disorders, six tests of interference and four tests of motor reactions were applied. The examinations were performed in 44 male workers exposed to Hg in the process of acetic aldehyde synthesis. The control group was matched for sex, age and educational level. The obtained results showed that in the detection od early dysfunctions of the nervous system, psychological tests of interference are more useful than test of the motor reactions. This toxicological test battery can be recommended for monitoring workers exposed to metallic mercury.