Abstract  The physicochemical and biological properties of metals change as the particles are reduced to nanoscale. This ability increases the application of nanoparticles in commercial and medical industry. Keeping in view this importance, Silver nanoparticles (Ag-NPs) were synthesized by reduction methods using formaldehyde as reducing agent in the chemical route and lemon extracts in the biological route. The scanning electron microscope (SEM) images of nanoparticles suggested that the particles were either agglomerated or spherical in shape with mean diameter of 16.59 nm in the chemical route and 42.93 nm in the biological route. The particles were between 5 and 80 nm with maximum frequency between 5 and 20 nm in the chemical route and between 5 and 100 nm with maximum frequency between 15 and 50 nm in the biological method. In the second phase of the study, the effect of Ag-NPs on the oxidative stress was studied. For this purpose, Labeo rohita (20 ± 2.5 g in weight and 12 ± 1.4 cm in length) were involved. Six treatments were applied in three replicates having five fishes in each replicate. The first treatment was used as control group, and the other five treatments were exposed to either 10 or 20 or 30 or 45 or 55 mg L-1 of Ag-NPs for 28 days. The treatment of Ag-NPs caused oxidative stress in the liver and gill tissues, which induced alterations in the activities of antioxidant enzymes. The level of catalase (CAT) was decreased in response to Ag-NPs concentration in dose-dependent manner. Ag-NPs treatment stimulated the liver and gill tissues to significantly increase the level of superoxide dismutase (SOD), which might be due to synthesis of SOD and addition in the pre-existing SOD level. The level decreases again due to depletion of SOD level. There was a sharp decline in the activities of glutathione S-transferase (GST) in both gills and liver tissues even at lower concentration, and this decrease in the GST activity was significantly different at each treatment after 28 days of treatment except 20 mg L-1. The malondialdehyde (MDA) levels of gills and liver tissues were increased with the increase in the concentration. The elevated levels of glutathione (GSH) showed that the liver started defensive mechanism against the oxyradicals. This study finds out the cheap eco-friendly and economical method of Ag-NP synthesis. It is further revealed that Ag-NPs caused oxidative stress in the aquatic animals if exposure occurs at high concentrations.

Abstract  OBJECTIVE: To investigate the effect of formaldehyde exposure on the expression of inflammatory cytokines in human bronchial epithelial cells (16HBE cells).

METHODS: 16HBE cells were treated with formaldehyde with a concentration of 0, 0.04, 0.08, 0.16, 0.32, or 0.64 mmol/L for 24 hours, and MTT assay was applied to measure proliferative activity and calculate median lethal dose; 16HBE cells were exposed to formaldehyde with a concentration of 0, 0.04, 0.16, 0.64, or 1.20 mmol/L for 4 hours, MTT assay was applied to measure proliferative activity, and enzyme-linked immunosorbent assay was applied to measure the levels of Th1, Th2, and Th17 cytokines and tumor necrosis factor α(TNF-α) in cell supernatant.

RESULTS: Compared with the control group, the 0.32-and 0.64-mmol/L exposure groups had significant decreases in cell viability (P<0.05); all exposure groups had reductions in interleukin(IL)-2 and IL-12, but no significant changes in interferon-γ and IL-10. In the 1.20-mmol/L exposure group, there was an increase in IL-4, with the increasing exposure dose, IL-5 and IL-6 tended to increase first and then decrease, and there was no significant change in IL-13; with the increasing exposure dose, IL-8 tended to increase first and then decrease, and there was no significant change in IL-17. In all the exposure groups, TNF-α increased and tended to increase significantly with the increasing exposure dose(P<0.05).

CONCLUSION: Formaldehyde exposure can cause imbalance between Th1 and Th2 cytokines secreted by 16HBE cells, as well as increased expression of IL-8 and TNF-α.

Abstract  Traditionally, chemical agents such as formalin (FA) and β-propiolactone (BPL) have long been used for the preparation of inactivated vaccines or toxoids. It has been shown that FA extensively modifies vaccine antigens and thus affects immunogenicity profiles, sometimes compromising the protective efficacy of the vaccines or even exacerbating the disease upon infection. In this study, we show that natural catechins from green tea extracts (GT) can be used as an inactivating agent to prepare inactivated viral vaccines. GT treatment resulted in complete and irreversible inactivation of influenza virus as well as dengue virus. In contrast to FA that reacted extensively with multiple amino acids including lysine, a major anchor residue for epitope binding to MHC molecules, GT catechin epigallocatechin-3-gallate (EGCG) crosslinked primarily with cysteine residues and thus preserved the major epitopes of the influenza hemagglutinin. In a mouse model, vaccination with GT-inactivated influenza virus (GTi virus) elicited higher levels of viral neutralizing antibodies than FA-inactivated virus (FAi virus). The vaccination completely protected the mice from a lethal challenge and restricted the challenge viral replication in the lungs. Of note, the quality of antibody responses of GTi virus was superior to that with FAi virus, in terms of the magnitude of antibody titer, cross-reactivity to hetero-subtypes of influenza viruses, and the avidity to viral antigens. As the first report of using non-toxic natural compounds for the preparation of inactivated viral vaccines, the present results could be translated into a clinically relevant vaccine platform with improved efficacy, safety, productivity, and public acceptance.

Abstract  The effects of using ultraviolet irradiation (UV), ozone (O-3) and the combined O-3-UV advanced oxidation process (AOP) on humic acid (HA), which is regarded as the main disinfection by-products (DBPs) precursor, have been evaluated and compared. In particular, aromatic acids, short-chain aldehydes and carboxylic acids were measured. The purpose of this study was to determine the different classes and yields of oxidation intermediates, compare O-3, UV and O-3-UV for HA removal, as well as investigate the effects of O-3, irradiation time, UV intensity on the mineralization of HA. Based on that, the knowledge gap of DBPs generation was made up. The results showed that by UV irradiation and O-3 oxidation, HA broke down into smaller molecules that were more hydrophilic, namely formaldehyde, acetaldehyde, propaldehyde, butyraldehyde, glyoxal, methyl-glyoxal, formic, acetic, fumaric, benzoic, phthalic, protocatechuic and 3-hydroxybenzoic acids. Meanwhile, unsaturated conjugated double bonds in the structure of HA were destroyed, which lead to UV254 a slight decrease. Due to the synergistic effect of O-3-UV, DOC and UV254 significantly decreased and remained stable after the reaction of 10min, indicating that O-3-UV system had stronger potential of mineralization and lower nonselectivity. Besides, the kinds and concentration levels of the intermediates were obviously reduced with light intensity increasing. Hydroxyl radical ((OH)-O-.) could mineralize some organics that could not be mineralized by O-3.

Abstract  OBJECTIVE: To observe the effect of different-doses of herbal cake-partitioned moxibustion (Moxi) on histopathological changes of the damaged colon tissue in rats with ulcerative colitis (UC), so as to select an optimal dosage of Moxi in the treatment of UC.

METHODS: Sixty-three male Sprague Dawley (SD) rats were randomized into 7 groups: normal control, model, medication, and 1, 2, 3 and 4 moxa-cone Moxi (n＝9 rats per group). The UC model was developed by subcutaneous injection of emulsion (1 mL) containing colon mucosa-prepared protein suspension and complete Freund's adjuvant into the toes, groin and back. On the 38th day, enema of 3% formalin and the aforementioned emulsion was used. Herbal-cake (composed of monkshood, cinnamon, etc.) partitioned Moxi with 1 or 2 moxa-cones (about 5 min/cone) was applied to bilateral "Tianshu" (ST 25) once daily or once every other day. The rat's general conditions (diet, movement, response ability, stool, and body weight) were observed, and histopathological changes (adhesion, ulcer formation and inflammation) of colon tissues were examined after hematoxylin-eosin (HE) staining, and scored (histopathological score). Gross score was given according to the severity of adhesion, ulcer formation and inflammation of colonic tissues under stereo microscope. The average optical density (AOD) values of colonic mucins were detected after periodic acid-schiff (PAS) staining, and those of the sulfated mucus content detected after high iron dia-mine-alcian blue (HID-AB) staining.

RESULTS: Compared with the normal group, rats in the model group presented loose stool, or with pus and blood, and slowly increased body weight (P<0.01), obvious congestion of colon with ulcer spots or continuous superficial ulcer regions which had irregular glandular cavities, swelling and serious inflammatory infiltration in lamina propria and submucosa, and increased score of colon tissue damage (P<0.01). PAS and HID-AB staining showed a marked decrease of AOD values of colonic mucins and sulfated mucus in the model group relevant to the normal control group (P<0.01), suggesting a reduction of mucus secretion of intestinal glands. Following the intervention, rats in the Moxi groups presented an increase of the body weight, formed feces, and an improvement of the damaged colon tissues as mucosal healing and inflammatory reduction, and a marked decrease of the damage score relevant to the model rats. No significant differences were found in the gross scores among the medication, 1, 2, 3 and 4 moxa-cone groups (P>0.05). The histopathological scores were significantly lower in the 1 and 4 moxa-cone Moxi groups than in the medication group (P<0.05, P<0.01); and significantly lower in the 1, 3, 4 moxa-cone Moxi groups than in the 2 moxa-cone Moxi group (P<0.05, P<0.01). PAS staining showed a significant increase of the AOD values of colonic mucins in the 1, 2, 3 and 4 moxa-cone and medication groups relevant to the model group (P<0.01); and the AOD values of colonic mucins in the 1, 3, 4 moxa-cone Moxi groups were significantly increased than that in the 2 moxa-cone Moxi group (P<0.05, P<0.01). HID-AB staining showed that the AOD values of sulfated mucus content were significantly higher in the 2 and 4 moxa-cone Moxi groups than in the 3 moxa-cone Moxi group (P<0.01). The two-level two-factor factorial analysis showed an interaction existed between the moxa-cone number and Moxi frequency in reducing the gross score and histopathological score and in facilitating colonic mucin and sulfated mucus secretion. The histopathological score of the 4 moxa-cone Moxi group was significantly lower than that of the 2 moxa-cone Moxi group (P<0.05), and the sulfated mucus content was significantly higher in the 4 moxa-cone group than in the 3 moxa-cone group (P<0.01). The effect of Moxi given on alternate days was superior to that of daily Moxi in improving colonic histological damage.

CONCLUSION: Herbal cake-partitioned moxibustion at ST 25 can promote repair of the damaged colonic tissue and secretion of mucin in UC rats. The number of moxa cones and intervention frequency affect the efficacy of Moxi in improving histopathological changes. The Moxi intervention on alternate days and with 2 moxa-cones every time is recommended.

Abstract  AIM: In this study, Epithelial Growth Factor Receptor and Platelet Endothelial Cell Adhesion Molecule-1 were localised to investigate the healing effects of a flavonoid-rich fraction of M. paradisiaca fruit in the gastric corpus of Wistar rats following aspirin-induced gastric lesion.

MATERIALS AND METHODS: Mature, unripe fruits of M. paradisiaca were peeled; air dried, pulverised, extracted with 70% methanol, concentrated and partitioned. Ninety male Wistar rats were randomly assigned into 6 groups of 15 rats each. The gastric lesion was induced in groups B, C, D, E and F rats by administration of 400 mg/kg aspirin in distilled water. Group A received distilled water. After 24 hours, flavonoid fraction of M. paradisiaca was administered to groups C, D and E at 100, 200 and 400 mg/kg respectively for 21 days. Group F rats received omeprazole at 1.8 mg/kg for 21 days. Five rats from each group were anaesthetized with ketamine on days 14, 21 and 28. Gastric tissues were excised and fixed in Neutral buffered formalin. This was followed by paraffin wax embedding method and sections stained with haematoxylin and eosin and for immunolocalisation of EGFR and PECAM-1. Data were analysed using descriptive and inferential statistics.

RESULTS: There was a significant difference in the ulcer index in the corpus of control and treated rats throughout the experimental period (p = 0.0001). H&E stained sections showed a gradual restoration of the epithelial lining in the treated groups. Immunohistochemical examination showed that M. paradisiaca significantly increased (p < 0.05) reactivity for both EGFR and CD31 across the treatment groups.

CONCLUSION: The efficacy of Musa paradisiaca in attenuating the damaging effects of aspirin on the gastric mucosa was observed as there was a significantly increased reactivity for EGFR and PECAM-1 in the gastric corpus in a dose-dependent manner.

Abstract  BACKGROUND: Pain is the most common cause of patients seeking medical advice as a result of its association with different pathologies. This study evaluated the antinociceptive property of Haematostaphis barteri as well as the possible mechanism(s) associated with its antinociceptive property.

METHODS: Mice were administered H. barteri (30-300 mg kg-1; p.o.), followed by intraplantar injection of 10 μL of 5% formalin into the hind paws. The pain score was determined for 1 h in the formalin test. The possible nociceptive pathways involved in the antinociceptive action of H. barteri were determined by pre-treating mice with theophylline (5 mg kg-1, a non-selective adenosine receptor antagonist), naloxone (2 mg kg-1, a non-selective opioid receptor antagonist), glibenclamide (8 mg kg-1; an ATP-sensitive K+ channel inhibitor), and atropine (3 mg kg-1; non-selective muscarinic antagonist).

RESULTS: H. barteri (30-300 mg kg-1) significantly and dose dependently precluded both first and second phases of nociception. Pre-treatment with naloxone had no effect on the analgesic activities of H. barteri in the first phase. Again, pre-treatment with atropine and glibenclamide did not significantly reverse the neurogenic antinociception of the extract in phase 1. However, theophylline reversed the analgesic effect of the extract in the first phase. In phase 2, theophylline had no effect on the analgesic activities of the extract. Naloxone, atropine, and glibenclamide significantly blocked the antinociception of H. barteri in the inflammatory phase of the formalin test.

CONCLUSIONS: H. barteri possesses antinociceptive property mediated via the opioidergic, adrenergic, muscarinic, ATP-sensitive K+ channels, and adenosinergic nociceptive pathways.

Abstract  Properties and mechanisms of PCNA (proliferating cell nuclear antigen) functions have been investigated for a long time and are studied in great detail. As follows from its name, most known PCNA functions (DNA replication, DNA repair, DNA recombination and others) are connected with cell proliferation and localization of this protein in nuclei. In addition, there is good reason to believe that PCNA also performs some functions in the cytoplasm. However, the possible role and mechanisms of PCNA action in the cytoplasm require careful study and clarification. Interestingly, such cells as neutrophils differ in that they are non-dividing on one hand and on the other hand contain a rather large amount of PCNA, which is localized only in the cytoplasm, that is, they are an ideal model for the study of cytoplasmic PCNA. Using cross-linkages with formaldehyde, we showed that this cytoplasmic PCNA is cross-linked in a similar way, that is, organized in the same way as the nuclear PCNA that is present in the proliferating cells. Previously, we showed that PCNA in such cells is organized into a dynamic complex of double trimer on the basis of the back-to-back principle (Naryzhny S.N. et al. (2005) J. Biol. Chem., 280, 13888). Apparently, such organization of this hub-protein allows it to better coordinate the processes taking place in the cytoplasm as well.

Abstract  In vitro cell culture studies showed that the hormonal form of vitamin D3, 1α,25-dihydroxyvitamin D3, significantly (p < 0.05) affects the human epigenome at thousands of genomic loci. Phase II of the VitDbol vitamin D intervention trial (NCT02063334) involved a proof-of-principle study of one individual, who was exposed three times every 28 days to an oral bolus (2000 μg) of vitamin D3. Blood samples were taken directly before each supplementation as well as one and two days after, chromatin was isolated from peripheral blood mononuclear cells without any further in vitro culture and at all nine time points epigenome-wide chromatin accessibility was assessed by applying FAIRE-seq (formaldehyde-assisted isolation of regulatory elements sequencing). The vitamin D3 bolus resulted in an average raise in 25-hydroxyvitamin D3 (25(OH)D3) serum concentration of 11.9 and 19.4 nM within one and two days, respectively. Consistently accessible chromatin was detected at 5205 genomic loci, the 853 most prominent of which a self-organizing map algorithm classified into early, delayed and non-responding genomic regions: 70 loci showed already after one day and 361 sites after two days significant (p < 0.0001) chromatin opening or closing. Interestingly, more than half of these genomic regions overlap with transcription start sites, but the change of chromatin accessibility at these sites has no direct effect on the transcriptome. Some of the vitamin D responsive chromatin sites cluster at specific loci within the human genome, the most prominent of which is the human leukocyte antigen region in chromosome 6. In conclusion, this study demonstrates that under in vivo conditions a rather minor rise in 25(OH)D3 serum levels is sufficient to result in significant changes at hundreds of sites within the epigenome of human leukocytes.

Abstract  Well-defined polyhedral Cu2O exposing different crystal facets are synthesized via a facile solution-phase method, and the facet-dependent gas sensing properties are investigated. The results show that cubic Cu2O exposing (100) facets has higher sensitivity to NO2, acetone and benzene than octahedral Cu2O with (111) facets. More importantly, the cubic Cu2O has high selectivity to NO2 and acetone at varied working temperatures by different sensing mechanisms of adsorption, chemical reaction, and intrinsic carrier excitation, as verified by the catalytic and electric property measurements. The higher sensitivity and selectivity of cubic Cu2O is explained by the relatively lower work function of (100) surfaces than that of (111), which induces a thicker surface hole accumulation layer and thus, a higher bulk hole activation energy (0.55eV and 0.36eV respectively). The synergistic effect of catalytic and electric properties gives rise to gas sensitivity decrease with the reduced percentage of (111) facets, which is beneficial to the gas sensing material design and also, is promising for the future highly sensitive and selective gas sensor fabrication.

Abstract  Numerical simulations were performed to assess the relationship between carbon monoxide (CO) and formaldehyde and unburned methanol-unregulated emissions in cylinders and a tailpipe with ozone (O-3) addition from a direct-injection spark-ignition methanol engine. This simulation study was performed during cold-start and steady-state modes with O-3 addition of 3000 and 7000 ppm. The initial phase of produced CO and formaldehyde is advanced significantly with O-3 addition, but the initial phase of produced unburned methanol had little impact. The effects of O-3 addition on the formation and oxidation of CO, formaldehyde and unburned methanol are lower for the steady-state compared with the cold-start mode. The effects of O-3 addition on CO, formaldehyde and unburned methanol production and consumption in the cylinder are formaldehyde > CO > unburned methanol. CO, formaldehyde and unburned methanol emissions decrease with increasing O-3 addition. When the exhaust valve opened, CO, formaldehyde and unburned methanol emissions with 7000 pprn O-3 addition for the cold-start mode are 153%, 52% and 70% lower than those without O-3 addition, respectively. CO, formaldehyde and unburned methanol emissions with 7000 ppm O-3 addition for the steady-state mode are 52.6%, 28% and 28% lower firan those without O-3 addition, respectively. (C) 2017 Elsevier Ltd. All rights reserved.

Abstract  AIM: The aim of this study was to examine the activation of neuronal Kv7/KCNQ channels by a novel modified Kv7 opener QO58-lysine and to test the anti-nociceptive effects of QO58-lysine on inflammatory pain in rodent models.

METHODS: Assays including whole-cell patch clamp recordings, HPLC, and in vivo pain behavioral evaluations were employed.

RESULTS: QO58-lysine caused instant activation of Kv7.2/7.3 currents, and increasing the dose of QO58-lysine resulted in a dose-dependent activation of Kv7.2/Kv7.3 currents with an EC50 of 1.2±0.2 μmol/L. QO58-lysine caused a leftward shift of the voltage-dependent activation of Kv7.2/Kv7.3 to a hyperpolarized potential at V1/2=-54.4±2.5 mV from V1/2=-26.0±0.6 mV. The half-life in plasma (t1/2) was derived as 2.9, 2.7, and 3.0 h for doses of 12.5, 25, and 50 mg/kg, respectively. The absolute bioavailabilities for the three doses (12.5, 25, and 50 mg/kg) of QO58-lysine (po) were determined as 13.7%, 24.3%, and 39.3%, respectively. QO58-lysine caused a concentration-dependent reduction in the licking times during phase II pain induced by the injection of formalin into the mouse hindpaw. In the Complete Freund's adjuvant (CFA)-induced inflammatory pain model in rats, oral or intraperitoneal administration of QO58-lysine resulted in a dose-dependent increase in the paw withdrawal threshold, and the anti-nociceptive effect on mechanical allodynia could be reversed by the channel-specific blocker XE991 (3 mg/kg).

CONCLUSION: Taken together, our findings show that a modified QO58 compound (QO58-lysine) can specifically activate Kv7.2/7.3/M-channels. Oral or intraperitoneal administration of QO58-lysine, which has improved bioavailability and a half-life of approximately 3 h in plasma, can reverse inflammatory pain in rodent animal models.

Abstract  Although Alzheimer's disease (AD) was first described over 100 years ago, there is still no suitable biomarker for diagnosing AD in easily collectable samples (e.g., blood plasma, saliva, and urine). Here, we investigated the relationship between morning urine formaldehyde concentration and cognitive impairment in patients with post-stroke dementia (PSD) or AD in this cross-sectional survey for 7 years. Cognitive abilities of the study participants (n = 577, four groups: 231 controls, 61 stroke, 65 PSD, and 220 AD) were assessed by Mini-Mental State Examination (MMSE). Morning urine formaldehyde concentrations were measured by high performance liquid chromatography (HPLC). Gender- and age-matched participants were selected from the four groups (n = 42 in each group). Both semicarbazide-sensitive amine oxidase (SSAO, a formaldehyde-generating enzyme) and formaldehyde levels in the blood and urine were analyzed by using an enzyme-linked immunosorbent assay (ELISA) and HPLC, respectively. We found that morning urine formaldehyde levels were inversely correlated with MMSE scores. The threshold value (the best Cut-Off value) of formaldehyde concentration for predicting cognitive impairment was 0.0418 mM in patients with PSD (Sensitivity: 92.3%; Specificity: 77.1%), and 0.0449 mM in patients with AD (Sensitivity: 94.1%; Specificity: 81.8%), respectively. The results of biochemical analysis revealed that the observed increase in urine formaldehyde resulted from an overexpression of SSAO in the blood. The findings suggest that measuring the concentration of formaldehyde in overnight fasting urine could be used as a potentially noninvasive method for evaluating the likelihood of ensuing cognitive impairment or dementia.

Abstract  This study was performed to assess attribution of high grade cervical intraepithelial neoplasia (HG-CIN) and invasive cervical cancer (ICC) to human papillomavirus (HPV) genotypes and secondarily to assess reproducibility of HG-CIN/ICC diagnosis obtained in Poland. Formaldehyde fixed, paraffin embedded blocks of HG-CIN/ICC from two distant institutions were sent to a central laboratory together with original histological diagnoses. Central/expert review of histopathological specimens was performed and agreement between local and central/expert diagnoses was calculated. HPV detection and genotyping in the samples was carried out with the use of SPF10-LiPA25 technology. Results were analyzed for 205 HG-CIN and 193 ICC cases with centrally confirmed diagnoses. Kappa coefficients and 95 % confidence intervals for HG-CIN and ICC diagnoses were: 0.13 (0.09;0.17) and 0.19 (0.11;0.26) respectively. Cohen's kappa coefficients for lesions with representative number of samples ranged from 0.01 for cervical intraepithelial neoplasia grade 2 to 0.75 for adenocarcinoma. HPV DNA was detected in 96.1 and 91.2 % of the confirmed HG-CIN and ICC specimens respectively. HPV positive HG-CIN was most commonly attributed to HPV types: 16 (62.8), 33 (7.8), 31 (6.6), 52 (3.7), 45 (2.6) and 58 (2.6 %). HPV positive ICC was most commonly attributed to HPV types: 16 (72.1), 18 (10.8), 33 (5.7), 45 (3.4) and 31 (1.7 %). Reproducibility of histological diagnosis of HG-CIN/ICC obtained in Poland generally increases with the severity of lesion and is lowest for cervical intraepithelial neoplasia grade 2 and highest for adenocarcinoma. Over 80 % of ICC cases are vaccine-preventable in Poland.

Abstract  OBJECTIVE: The objective of this study was to determine the frequency and anatomical characteristics of Struthers' ligament and the supracondylar humeral process and evaluate the clinical implications in compressive neuropathy of the median nerve .

METHOD: We dissected 60 arms from 30 cadavers (26 males and 4 females): 15 were previously preserved in formalin and glycerin and 15 were dissected fresh in the Anatomy Laboratory for this paper. The relationships between Struthers' ligament and the median nerve and brachial artery and veins were documented with drawings and photos .

RESULTS: The supracondylar humeral process was not found in any of the 60 dissected arms. Struthers' ligament was identified in six arms (two bilateral); in all cases high insertion of the pronator teres muscle was observed .

CONCLUSION: Struthers' ligament is an aponeurotic structure that may or may not be associated with the supracondylar humeral process, and is an important potential site of median nerve compression in the lower third of the arm. Level of Evidence IV, Case Series.

Abstract  In this study, volume densities of white and grey matters of cervical segments of spinal cords of quail were investigated stereologically. In both sexes, mature, six quails were used as material of this study. All animals were fixed by perfusing in 10% buffered formalin. Tissue specimens were obtained from cervical spinal cords. These tissue specimens were cut every fiftieth section at 5 μm thickness by a microtome. And mean six or seven sections were examined from every block by this method at microscope. After that, these sections were stained by haematoxylin eosin and photographed. Densities of volumes of all tissues of cervical segments of whole spinal cords and white and grey matters were calculated with principle of Cavalieri. As a result, total volume of spinal cord, volumes of white and grey matters of cervical segment and volume rates of white and grey matters were calculated.

Abstract  While mechanochemical transduction principles are omnipresent in nature, mimicking these in artificial materials is challenging. The ability to reliably detect the exposure of man-made objects to mechanical forces is, however, of great interest for many applications, including structural health monitoring and tamper-proof packaging. A useful concept to achieve mechanochromic responses in polymers is the integration of microcapsules, which rupture upon deformation and release a payload causing a visually detectable response. Herein, it is reported that this approach can be used to create mechanochromic fluorescent materials that show a direct and ratiometric response to mechanical deformation. This can be achieved by filling poly(urea-formaldehyde) microcapsules with a solution of a photoluminescent aggregachromic cyano-substituted oligo(p-phenylene vinylene) and embedding these particles in poly(dimethylsiloxane). The application of mechanical force by way of impact, incision, or tensile deformation opens the microcapsules and releases the fluorophore in the damaged area. Due to excimer formation, the subsequent aggregation of the dye furnishes a detectable fluorescence color change. With the emission from unopened microcapsules as built-in reference, the approach affords materials that are self-calibrating. This new concept appears to be readily applicable to a range of polymer matrices and allows for the straightforward assessment of their structural integrity.

Abstract  BACKGROUND: The pterional approach (PA), together with its variants, is still one of the most common methods used by surgeons to reach the anterior and middle cranial base. A highly important technical detail during a PA is the preservation of the frontotemporal branch of the facial nerve, which can be achieved through an interfascial dissection.

OBJECTIVE: To describe the anatomy of the interfascial vein (IFV), highlighting its recognition as a significant anatomic reference to perform an interfascial dissection (IFD).

METHODS: Eight adult cadaveric heads, fixed with formaldehyde and injected with colored silicone, were studied. In 6 heads, an IFD was performed, simulating a PA. In the 2 remaining heads, the IFV was dissected. In addition, an IFD was performed in 10 patients, studying the IFV anatomy.

RESULTS: In the 6 cadaveric heads in which the PA with an IFD was performed, and in the 10 patients who underwent a PA with an IFD, the IFV was found. If the interfascial space is divided into thirds, in all cases, the IFV was located within the middle third of the interfascial fat pad. On the 2 cadaveric heads in which the IFV was anatomically dissected, the IFV was also located within the middle third of the interfascial space.

CONCLUSION: Recognizing the IFV in the interfascial space is of great help as an anatomic landmark to confirm that one is actually between both layers of the superficial temporal fascia.

Abstract  The ornamental fish trade provides a pathway for the global translocation of aquatic parasites. We examined a total of 1020 fish imported from Singapore, Malaysia, Thailand, or Sri Lanka to Australia (including freshwater and marine fish species) for monogenean ectoparasites. Fish were received following veterinary certification that they showed no clinical signs of pests and diseases from the exporting country and visual inspection at Australian border control. Australian import conditions require mandatory treatment for goldfish with parasiticides (e.g. trichlorfon, formaldehyde, sodium chloride) for the presence of gill flukes (Dactylogyrus vastator Nybelin, 1924 and Dactylogyrus extensus Mueller and Van Cleave, 1932) prior to export. Over 950 individual parasites were detected in five imported fish species, representing 14 monogenean species. Seven Dactylogyrus spp. including D. vastator and three Gyrodactylus spp. infected goldfish, Carassius auratus Linnaeus, 1758, from Malaysia, Singapore, and Thailand. Dactylogyrus ostraviensis Řehulka, 1988, infected rosy barb, Pethia conchonius Hamilton, 1822, from Singapore, Sri Lanka, and Thailand while two Trianchoratus spp. infected three spot gourami, Trichopodus trichopterus Pallas, 1970 and pearl gourami Trichopodus leerii Bleeker, 1852, from Sri Lanka. Urocleidoides reticulatus Mizelle & Price, 1964, infected guppy, Poecilia reticulata Peters, 1859, from Sri Lanka. The discovery of D. vastator in goldfish, as well as 13 other monogenean species, shows that pre-export health requirements, which include chemical treatment of goldfish, and inspection of all ornamental fish species did not prevent infection by monogeneans. Inspection prior to exportation and at border control must account for the highly cryptic nature of monogenean parasites and consider alternatives to current pre-export conditions and visual inspection at border control.

Abstract  K(+)-Cl(-) co-transporter (KCC2) and Na(+)-K(+)-2Cl(-) co-transporter (NKCC1) are the main regulators of neuronal intracellular chloride concentration; altered expression patterns of KCC2 and NKCC1 have been reported in several neurodegenerative diseases. In this paper, we show the effect of repeated stress on KCC2, NKCC1, and serine 940 phosphorylated KCC2 (pKCC2(ser940)) immunoreactivity. The data were obtained from the hippocampus of female mice using single-plane confocal microscopy images. The mean fluorescence intensity of the perisomatic area of neurons, defined as raw fluorescence intensity (RFI) was calculated. Repeated stress (RS) resulted in a decrease in perisomatic area of immunoreactive (IR)-KCC2 and an increase of the IR-NKCC1. In addition, RS decreased perisomatic IR-pKCC2(ser940), corresponding to that of KCC2. The data in this article support the results of a previous study [1] and provide the details of immunohistological methods. Interpretation of the data in this article can be found in "Repeated stress-induced expression pattern alterations of the hippocampal chloride transporters KCC2 and NKCC1 associated with behavioral abnormalities in female mice" by Tsukahara et al. [1].

Abstract  Brain-derived neurotrophic factor (BDNF) is released from activated microglia during neuropathic pain and is hypothesized to downregulate the expression of the potassium chloride cotransporter 2 (KCC2) via the TrkB receptor. Previous studies reported that KCC2 is downregulated 5 min after the plantar injection of formalin in rats; however, the mechanism behind this decrease in KCC2 expression during acute inflammatory pain remains unknown. In this study, we determined whether the TrkB receptor contributes to the expression of KCC2 during the acute pain. Five minutes after the plantar injection of formalin in rats, the ratio of KCC2-immunoreactive area in layer II of the spinal cord significantly decreased on the stimulated side compared to the unaffected side. On the other hand, this response was inhibited by the injection of a kinase inhibitor, K252a, in the subarachnoid space 15 min before the formalin injection. These findings suggest that in acute pain, the TrkB receptor may contribute to the decrease in the expression of KCC2.

Abstract  This study aimed to clarify the genomic factors associated with the diagnosis and prognosis of oral squamous cell carcinoma via next-generation sequencing. We evaluated data from 220 cases of oral squamous cell carcinoma. Genomic DNA was eluted using formalin-fixed, paraffin-embedded samples, and targeted resequencing of 50 cancer-related genes was performed. In total, 311 somatic mutations were detected in 220 patients, consisting of 68 synonymous mutations and 243 non-synonymous mutations. Genes carrying mutations included TP53, CDKN2A, and PIK3CA in 79 (35.9%), 35 (15.9%), and 19 patients (8.6%), respectively. Copy number analysis detected amplification of PIK3CA and AKT1 in 38 (17.3%) and 11 patients (5.0%), respectively. Amplification of receptor tyrosine kinases was found in 37 patients (16.8%). Distant metastasis was noted in nine of 37 patients (24%) with receptor tyrosine kinase amplification, accounting for 43% of the 21 cases of distant metastasis. The cumulative 5-year survival rate was 64.6% in the receptor tyrosine kinase amplification group vs 85.2% in the no receptor tyrosine kinase amplification group. Moreover, we identified significantly poorer prognosis in the TP53 mutation/receptor tyrosine kinase amplification group, for which the cumulative 5-year survival rate was 41.6%. In conclusion, the results of this study demonstrated that receptor tyrosine kinase amplification is a prognostic factor for distant metastasis of oral squamous cell carcinoma, indicating the necessity of using next-generation sequencing in clinical sequencing.

Abstract  Archival formalin-fixed, paraffin-embedded (FFPE) tumor specimens were collected from advanced NSCLC patients enrolled in LETS phase III trial comparing first-line S-1/carboplatin with paclitaxel/carboplatin and subjected to multiplex genotyping for 214 somatic hotspot mutations in 26 genes (LungCarta Panel) and 20 major variants of ALK, RET, and ROS1 fusion genes (LungFusion Panel) with the Sequenom MassARRAY platform. MET amplification was evaluated by fluorescence in situ hybridization. A somatic mutation in at least one gene was identified in 48% of non-squamous cell carcinoma and 45% of squamous cell carcinoma specimens, with EGFR (17%), TP53 (11%), STK11 (9.8%), MET (7.6%), and KRAS (6.2%). Mutations in EGFR or KRAS were associated with a longer or shorter median overall survival, respectively. The LungFusion Panel identified ALK fusions in six cases (2.5%), ROS1 fusions in five cases (2.1%), and a RET fusion in one case (0.4%), with these three types of rearrangement being mutually exclusive. Nine (3.9%) of 229 patients were found to be positive for de novo MET amplification. This first multiplex genotyping of NSCLC associated with a phase III trial shows that MassARRAY-based genetic testing for somatic mutations and fusion genes performs well with nucleic acid derived from FFPE specimens of NSCLC tissue.

Abstract  Along with the well-known rewarding effects, activation of nicotinic acetylcholine receptors (nAChRs) can also relieve pain, and some nicotinic agonists have analgesic efficacy similar to opioids. A major target of analgesic drugs is the descending pain modulatory pathway, including the ventrolateral periaqueductal gray (vlPAG) and the rostral ventromedial medulla (RVM). Although activating nAChRs within this circuitry can be analgesic, little is known about the subunit composition and cellular effects of these receptors, particularly within the vlPAG. Using electrophysiology in brain slices from adult male rats, we examined nAChR effects on vlPAG neurons that project to the RVM. We found that 63% of PAG-RVM projection neurons expressed functional nAChRs, which were exclusively of the α7-subtype. Interestingly, the neurons that express α7 nAChRs were largely nonoverlapping with those expressing μ-opioid receptors (MOR). As nAChRs are excitatory and MORs are inhibitory, these data suggest distinct roles for these neuronal classes in pain modulation. Along with direct excitation, we also found that presynaptic nAChRs enhanced GABAergic release preferentially onto neurons that lacked α7 nAChRs. In addition, presynaptic nAChRs enhanced glutamatergic inputs onto all PAG-RVM projection neuron classes to a similar extent. In behavioral testing, both systemic and intra-vlPAG administration of the α7 nAChR-selective agonist, PHA-543,613, was antinociceptive in the formalin assay. Furthermore, intra-vlPAG α7 antagonist pretreatment blocked PHA-543,613-induced antinociception via either administration method. Systemic administration of submaximal doses of the α7 agonist and morphine produced additive antinociceptive effects. Together, our findings indicate that the vlPAG is a key site of action for α7 nAChR-mediated antinociception.

Abstract  BACKGROUND: The role of internal mammary lymph node biopsy (IMLNB) is still being discussed in breast cancer treatment. The aim of this study was to investigate the role of IMLNB on adjuvant therapy and survival of patients with breast cancer.

PATEINTS AND METHODS: The data of 72 patients with clinically negative axilla and IMLNB were evaluated. IMLNB was performed either through a small separate intercostal incision or from the same incision for tumor resection or mastectomy by using both blue dye and radioisotope. Pathological analysis was performed on formalin-fixed paraffin-embedded tissues.

RESULTS: Ten of the patients (14%) were IMLNB-positive. The axillary sentinel lymph node and IMLN were negative in most of the patients (52.8%). In one patient (1.4%), the axilla was negative but the IMLNB was positive. IMLNB changed the pathologic stage in eight patients (11%). Adjuvant internal mammary radiotherapy was added to the treatment protocol for 10 patients due to IMLNB positivity and adjuvant chemotherapy was added in for only one patient with negative axilla. The factors found to be related with IMLN positivity were SLN positivity (p = 0.033), mastectomy (p = 0.022), and the number of resected IMLN ≥2 (p = 0.040). The median follow-up time was 115.5 months (range, 30-162 months). The ten-year overall survival (OS) rate was 86%. Systemic metastasis (p = 0.007), SLNB positivity (p < 0.001), and IMLNB positivity (p = 0.005) were statistically related to overall survival.

CONCLUSION: IMLNB positivity in patients with breast cancer changed the pathologic stage and adjuvant treatment modalities of patients and also adversely affected the overall survival.