Clinicopathological features and course of cutaneous protothecosis.

BACKGROUND: Protothecosis is an uncommon infection caused by the achlorophyllic algae found more commonly in tropical areas. Only a limited number of cases have been reported. OBJECTIVE: We aimed to evaluate the clinicopathological features and treatment outcomes of cutaneous protothecosis. METHODS: We retrospectively identified 20 pathology-confirmed cases of cutaneous protothecosis based on skin biopsies in two tertiary medical centres in Taiwan from 1997 to 2015. RESULTS: The age of the patients at the time of diagnosis ranged from 48 to 85 years (mean age of 74 years). All lesions developed on the limbs. Twelve (60%) patients had adrenal insufficiency, but no patients had active malignancy at diagnosis. Interestingly, four (20%) patients had concurrent scabies infestation. Clinically, most lesions were erythematous plaques studded with punctate ulcers. Microscopically, the most common finding was granulomatous inflammation. Nineteen (95%) cases were successfully treated with itraconazole for 14-148 days with only one case of recurrence. Concomitant scabies should be suspected if pruritus is recalcitrant despite itraconazole treatment. CONCLUSION: Despite its rarity, cutaneous protothecosis has become more significant due to an increased prevalence of immunocompromised individuals. Steroid overuse or iatrogenic adrenal insufficiency predisposes individuals to high-risk infections. Neglecting the disease leads to a chronic and incurable state. Protothecosis should be suspected in chronic eczematous and ulcerative plaques on the limbs refractory to conventional antibacterial and antiviral treatments, especially in patients with adrenal insufficiency. Clinical suspicion should be confirmed by skin biopsies, and confirmed cases can be successfully treated with itraconazole.

Effects of agricultural land use on fluvial carbon dioxide, methane and nitrous oxide concentrations in a large European river, the Meuse (Belgium).

We report a data-set of CO2, CH4, and N2O concentrations in the surface waters of the Meuse river network in Belgium, obtained during four surveys covering 50 stations (summer 2013 and late winter 2013, 2014 and 2015), from yearly cycles in four rivers of variable size and catchment land cover, and from 111 groundwater samples. Surface waters of the Meuse river network were over-saturated in CO2, CH4, N2O with respect to atmospheric equilibrium, acting as sources of these greenhouse gases to the atmosphere, although the dissolved gases also showed marked seasonal and spatial variations. Seasonal variations were related to changes in freshwater discharge following the hydrological cycle, with highest concentrations of CO2, CH4, N2O during low water owing to a longer water residence time and lower currents (i.e. lower gas transfer velocities), both contributing to the accumulation of gases in the water column, combined with higher temperatures favourable to microbial processes. Inter-annual differences of discharge also led to differences in CH4 and N2O that were higher in years with prolonged low water periods. Spatial variations were mostly due to differences in land cover over the catchments, with systems dominated by agriculture (croplands and pastures) having higher CO2, CH4, N2O levels than forested systems. This seemed to be related to higher levels of dissolved and particulate organic matter, as well as dissolved inorganic nitrogen in agriculture dominated systems compared to forested ones. Groundwater had very low CH4 concentrations in the shallow and unconfined aquifers (mostly fractured limestones) of the Meuse basin, hence, should not contribute significantly to the high CH4 levels in surface riverine waters. Owing to high dissolved concentrations, groundwater could potentially transfer important quantities of CO2 and N2O to surface waters of the Meuse basin, although this hypothesis remains to be tested.

A tryptophan metabolite, kynurenine, promotes mast cell activation through aryl hydrocarbon receptor.

BACKGROUND: Tryptophan metabolites have been suggested to play a role in immune modulation, wherein those have recently been shown to be endogenous ligands of aryl hydrocarbon receptor (AhR; a unique cellular chemical sensor). However, the involvement of tryptophan metabolites and AhR in modulating mast cell function remains to be fully defined. We therefore investigated that the functional impacts of tryptophan metabolites on human and mouse mast cell responses in vitro and their functional importance in vivo. METHODS: Three tryptophan metabolites, kynurenine (KYN), kynurenic acid (KA) and quinolinic acid (QA), were examined in terms of their effect on IgE-mediated responses in mouse bone marrow-derived mast cells (BMMCs) and in human peripheral blood-derived cultured mast cells (HCMCs) and on in vivo anaphylactic responses. For evaluation of AhR involvement, we examined the responses of mast cells from AhR-null or AhR-wild-type mice with the use of a known AhR antagonist, CH223191. RESULTS: Kynurenine, but not KA and QA, enhanced IgE-mediated responses, including degranulation, LTC4 release, and IL-13 production in BMMCs through the activation of PLCγ1, Akt, MAPK p38, and the increase of intracellular calcium. KYN also enhanced cutaneous anaphylaxis in vivo. These enhancing effects of KYN were not observed in AhR-deficient BMMCs and could be inhibited by CH223191 in BMMCs. Further, KYN had similar enhancing effects on HCMCs, which were inhibited by CH223191. CONCLUSION: The AhR-KYN axis is potentially important in modulating mast cell responses and represents an example of AhR's critical involvement in the regulation of allergic responses.

Work-related chemical exposures presenting to an emergency department in Singapore.

BACKGROUND: Singapore is a small industrialized island state with a low accident rate and few hazardous chemical-related injuries reported. However, the use of chemicals continues to increase and pose hazards in the workplace. AIMS: To study workplace chemical injuries and exposures to improve worksite safety. METHODS: Work-related chemical exposure cases were identified from emergency department (ED) computerized records from 2007 to 2010. RESULTS: A total of 239 cases were identified. Most of the patients were male (92%) and young adults (73% aged between 21 and 40 years). Fifty per cent of the workers were foreign workers. Most of them were cleaners, labourers and technicians (53%) and worked mainly in the construction and manufacturing industries (47%). All the exposures were acute and presented within 4h of the exposure incident (52%). Most of the chemical exposures were to the eye (55%) and skin (32%). The chemicals involved included corrosives (41%), hydrocarbons (18%) and cleaning solutions (9%). Pre-hospital decontamination (eye and skin irrigation) was performed for 54% of the workers. Antidote treatment with calcium gluconate for hydrofluoric acid exposure was used for five patients in the ED. Only 11% of patients were admitted. Four patients had surgical procedures and five patients had long-term complications. Forty-five incidents were notified to the Ministry of Manpower. The under-reporting rate for cases with >3 days of medical leave was 66%. CONCLUSIONS: Work-related chemical exposures that present to the ED had low morbidity. Most of the workers did well with immediate decontamination and supportive treatment but antidotes were required for some exposures.

Impact of neck dissection in early tongue and buccal cancer without neck extension.

PROBLEM: The role of elective neck dissection in early stage tongue and buccal squamous cell carcinoma with negative neck lymph nodes is still controversial. METHODS: We retrospectively reviewed patients with T1-2N0M0 buccal and tongue cancer who underwent primary tumour excision with or without elective neck dissection between January 1997 and December 2006. RESULTS: Elective neck dissection specifically improved disease-free survival of T2N0M0 buccal cancer and overall survival of T2N0M0 tongue cancer. CONCLUSION: Elective neck dissection seems to improve the disease-free survival rate of T2N0M0 buccal cancer and the overall survival rate of T2N0M0 tongue cancer but has no beneficial effect on the survival rate of T1N0M0 buccal and tongue cancer.

Intracellular zinc release-activated ERK-dependent GSK-3ß-p53 and Noxa-Mcl-1 signaling are both involved in cardiac ischemic-reperfusion injury.

Oxidative stress and nitrosative stress are both suggested to be involved in cardiac ischemia-reperfusion (I/R) injury. Using time-lapse confocal microscopy of cardiomyocytes and high-affinity O(2)(-•) and Zn(2+) probes, this study is the first to show that I/R, reactive oxygen species (ROS), and reactive nitrogen species (RNS) all cause a marked increase in the [O(2)(-•)](i), resulting in cytosolic and mitochondrial Zn(2+) release. Exposure to a cell-penetrating, high-affinity Zn(2+)(i) chelator, TPEN, largely abolished the Zn(2+)(i) release and markedly protected myocytes from I/R-, ROS-, RNS-, or Zn(2+)/K(+) (Zn(2+)(i) supplementation)-induced myocyte apoptosis for at least 24 h after TPEN removal. Flavonoids and U0126 (a MEK1/2 inhibitor) largely inhibited the myocyte apoptosis and the TPEN-sensitive I/R- or Zn(2+)(i) supplement-induced persistent extracellular signal-regulated kinase 1 and 2 (ERK1/2) phosphorylation, dephosphorylation of p-Ser9 on glycogen synthase kinase 3ß (GSK-3ß), and the translocation into and accumulation of p-Tyr216 GSK-3ß and p53 in, the nucleus. Silencing of GSK-3ß or p53 expression was cardioprotective, indicating that activation of the ERK-GSK-3ß-p53 signaling pathway is involved in Zn(2+)-sensitive myocyte death. Moreover, the ERK-dependent Noxa-myeloid cell leukemia-1 (Mcl-1) pathway is also involved, as silencing of Noxa expression was cardioprotective and U0126 abolished both the increase in Noxa expression and in Mcl-1 degradation. Thus, acute upstream Zn(2+)(i) chelation at the start of reperfusion and the use of natural products, that is, flavonoids, may be beneficial in the treatment of cardiac I/R injury.

Flow density of a reversible ratchet.

By defining "flow density" in a reversible ratchet, a kind of Brownian motor that runs four processes in a two-dimensional parameter space to complete a cycle, we analytically investigate the behavior of the motor's integrated flow in terms of the magnitude and location of the density maximum. It turns out that flow density gives information that can serve as a guide for obtaining the most suitable integrated flow important in the design of reversible ratchets.

Comparison of invasive and non-invasive measurement of spontaneous baroreflex during anesthesia.

BACKGROUND: Many works have shown that beat-to-beat variations in systolic arterial pressure (SAP) could be used for the assessment of cardiac baroreflex function. The purpose of this study was to evaluate whether Finapres, a non-invasive device to measure systolic arterial pressure, could serve as an acceptable alternative to intra-arterial catheterization for assessing the spontaneous baroreflex during anesthesia. METHODS: Thirty ASA class I patients scheduled for surgical hepatic and gastric tumor excisions were enrolled in this study. Spontaneous baroreflex sensitivity (SBR) during anesthesia was calculated by sequence analysis of beat-to-beat variations in systolic arterial pressure and R-R intervals. Data obtained from Finapres were compared with those from standard intra-arterial catheterization technique. RESULTS: SBR values determined by non-invasive and invasive blood pressure measurement technique were precisely correlated (r = 0.96) and the mean difference was about 0.22 ms/mmHg (12.5%). However, the individual data might differ by -0.52 to +0.96 ms/mmHg (-35% to +60%). CONCLUSIONS: Finapres may trace the changes of SBR during anesthesia and provides similar results as does invasive method but individual variation exists.

Transesophageal echocardiographic monitoring for transcatheter closure of atrial septal defect.

BACKGROUND AND PURPOSE: Transcatheter closure of atrial septal defect (ASD) is generally performed under fluoroscopy alone. Recently, we have used transesophageal echocardiography (TEE) monitoring as an aid in performing this procedure. The purpose of this study was to evaluate the efficacy and complications associated with this use of TEE. METHODS: Transcatheter closure of ASD was accomplished under TEE guidance simultaneously with fluoroscopic imaging in 11 patients aged 3 to 33 years (weight, 15.4-62.9 kg). TEE was successfully performed in all patients after endotracheal general anesthesia. The ASDs were reexamined before catheterization. The appropriate placement of the occluder device was evaluated. RESULTS: Seven cases were uneventful with successful ASD occluder implantation, but one failed because of a large ASD (24.7 mm). In three cases, transcatheter closure was aborted after TEE examination, one with a large ASD (27.05 mm), one with an ASD that was too small, and one with multiple fenestrated ASDs. CONCLUSIONS: Routine TEE monitoring for transcatheter closure of ASDs is effective for evaluation of ASD before implantation of an occluder, to ensure the proper seating of the occluder after the defect occlusion is complete.

Spinal anesthesia in MELAS syndrome: a case with mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes.

MELAS syndrome (mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes) is one of the classic mitochondrial encephalomyopathies with variable clinical presentation and multisystem involvement. Enhanced sensitivity to neuromuscular blockade or anesthetic agents and susceptibility to malignant hyperthermia in these patients have ever been reported, all of which complicate the management of general anesthesia. To avoid these appalling troubles in general anesthesia, we chose spinal anesthesia for a patient with MELAS syndrome receiving appendectomy. The patient obtained adequate anesthesia and good recovery without neurologic sequelae. Although there is little information about the application of regional anesthesia in MELAS patients, we demonstrate that it may be a satisfactory choice. However, it is suggested that regional anesthesia is performed only when neurological abnormalities of spinal cord or peripheral nerves are definitely ruled out.

Phosphorylated tau can promote tubulin assembly.

Phosphorylation can affect the function of microtubule-associated protein tau. Here, the human brain tau with 441 amino acids was phosphorylated by cyclic-AMP-dependent protein kinase (PKA) or glycogen synthase kinase-3beta. PKA-phosphorylated tau (2.7 mol phosphates/mol) does not promote tubulin assembly as judged by spectrophotometric and atomic force microscopy measurements, unless trimethylamine N-oxide (TMAO), a natural occurring osmolyte, is included in these assays. TMAO is also found to promote tubulin assembly of glycogen synthase kinase-3beta-phosphorylated tau (1.6 mol phosphates/mol). TMAO does not act by causing a chemical dephosphorylation of phosphorylated tau, but it acts to overcome the functional deficit caused by phosphorylation. PKA-phosphorylated tau binds to tubulin in the presence of TMAO and lowers the critical concentration of tubulin needed for assembly. From these data, we conclude that PKA-phosphorylated tau retains the ability to bind tubulin and promote tubulin assembly. TMAO is required, however, to sensitize the reaction. Possible uses of TMAO in relation to studies of tubulin assembly in vitro, in intact cells, and in relation to Alzheimer's disease are presented in this report.

Hepatobiliary dysfunction as the initial manifestation of disseminated cryptococcosis.

A case of hepatobiliary dysfunction as the initial manifestation of disseminated cryptococcosis is described. The patient was admitted with symptoms of hepatitis with cholestatic jaundice. Antibody tests for hepatitis B and C and human immunodeficiency virus were negative. The patient continued to deteriorate clinically. Eventually, the patient succumbed to hepatic failure. Autopsy disclosed systemic cryptococcosis that caused extensive necrosis of the liver. In review of the literature, only nine cases of cryptococcal infection presenting as hepatitis, cholangitis, and cholecystitis as initial manifestation were reported. Four of these patients had been subjected to exploratory laparotomy for clinical suspicion of acute abdomen. One patient developed cirrhosis as a result of cryptococcal hepatitis. Two patients succumbed to hepatic failure. Cryptococcosis is known to occur commonly in immunocompromised patients, yet only two reported cases presenting as hepatitis were associated with immunocompromised status.

Natural methylamine osmolytes, trimethylamine N-oxide and betaine, increase tau-induced polymerization of microtubules.

The natural osmolyte trimethylamine N-oxide (TMAO) at 200 mM increases the extent and the rate of formation of polymerized microtubule (MT) complex caused by tau. TMAO at this concentration has no effect on tubulin alone. Urea at 200 mM blocks tubulin assembly caused by tau, but this inhibition can be reversed by an equal amount of TMAO. Besides TMAO, betaine, another natural osmolyte, was found to have the same effects on MT as TMAO. On the contrary, glycerol (a carbohydrate osmolyte) and glycine (an amino acid osmolyte) do not increase tau-induced MT assembly. The mechanism by which TMAO and betaine enhance tau's effectiveness is not known, but physical studies suggest that the secondary structure of tau is not appreciably changed by 200 mM TMAO. This is the first report showing that natural osmolytes, TMAO and betaine, at a near physiological concentration are able to stimulate tau-induced tubulin assembly.

The commissioning of a flexible low-cost multipurpose X-ray beamline at SRRC.

A low-cost multipurpose X-ray beamline has been commissioned at the bending magnet B15 of the Synchrotron Radiation Research Center (SRRC). The beamline is constructed in such a way to accommodate the various needs of small research consortia, from universities or research institutes, whose requirements of a beamline facility are quite diverse while under limited funding support. Flexibility is the special feature of this beamline. It is capable of performing quick test measurements without a prolonged reviewing process. Switching between different techniques, such as white-beam irradiation, EXAFS and X-ray scattering, can be achieved within 1 h. Novel experiments, such as energy-dispersive small-angle scattering experiments and energy-dispersive reflectivity measurements, can also be performed.

The endocrine secretion of human insulin and growth hormone by exocrine glands of the gastrointestinal tract.

The exocrine pancreas, liver, and submandibular glands of the rat were used to express and secrete two exogenous, human protein hormones (growth hormone and insulin) into blood at physiological concentrations. Transfection, expression, and secretion were achieved by the in vivo retrograde injection of plasmid DNA into the secretory ducts of these glands. Pancreatic acinar cells secreted physiological concentrations of growth hormone into the circulation, and its secretion was enhanced by cholinergic stimulation. A human insulin gene was engineered to allow normal processing of insulin in non-beta cells. With this gene, the secretion of human insulin by the exocrine pancreas normalized elevated blood glucose levels in diabetic rats. These in vivo observations demonstrate the utility of retrograde ductal administration of naked DNA into exocrine organs as a novel method for the regulated systemic delivery of protein-based pharmaceuticals.

Comparison of stored and secreted rat pancreatic digestive enzymes by mass spectrometry: alpha-amylase.

As part of a continuing effort to better understand the mechanisms of protein secretion, we compared the mass of pancreatic digestive enzymes, in resting and stimulated states, both in secretion and in the zymogen granule to determine whether their secretion is accompanied by chemical modification. Mass spectra were obtained applying the electrospray method on samples separated by reverse-phase HPLC. We report here our results for alpha-amylase (1,4-alpha-D-glucan glucanohydrolase EC 3.2.1.1). The data illustrate structural differences between states and compartments for this enzyme. Multiple isozymes were identified from the mass spectra, varying roughly from 52 to 60 kDa. On the basis of mass comparisons, not all of the products seen in the zymogen granule were found in secretion, nor were all secreted isoforms in the granule. Stimulation of protein secretion with a cholinergic agonist, led to time-dependent changes in the number and masses of isoforms in secretion, leaving only one of five resolvable forms in the granule. Only one form, 55.5 kDa, was found in all samples, granule and secretion. In addition to these differences, microheterogeneities of 400 Da or less were observed. The data suggest the differential or non-parallel release of different amylase forms and their chemical modification during the secretion process. As such, release appears to involve a third, intermediate compartment, between zymogen granule to ductal space, such as the cytoplasm, in which chemical modification takes place.

Characterization of the manganese-resistant mutants derived from Vibrio parahaemolyticus.

The virulence and some related factors of Vibrio parahaemolyticus are regulated by the level of iron. In this study, five Mn-resistant mutants were selected after N-methyl-N'-nitrosoguanidine treatment and two transfers in medium containing high levels of manganese chloride. Production of siderophores and the 77-kDa iron-regulated outer-membrane protein and the bacterial growth in these Mn-resistants were deregulated, as compared with the wild-type strain. In addition, the regulation of these phenomena was partially or completely restored by the introduction of Escherichia coli fur gene. Also, the total cellular protein profiles of the wild-type and mutants showed that production of some proteins were positively or negatively regulated by iron, and expression of some of these proteins remained unaffected in these mutants. These results suggested the presence of a complicated iron regulation system, similar to the Fur system of E. coli, in this pathogen.

Selective determination of adenine-containing compounds by capillary electrophoresis with laser-induced fluorescence detection.

Capillary electrophoresis coupled with laser-induced fluorescence detection provides a selective analysis of mixtures of adenine-containing compounds (adenine, adenosine, cAMP, AMP, ADP, and ATP) that are derivatized using chloroacetaldehyde as a fluorogenic reagent. The components can be detected with linear response over the concentration range of 10(-4) to 10(-9) M, and the detection limit corresponds to 5 to 10 x 10(-19) mol for each nucleotide. Analysis is achieved in less than 10 min with high separation efficiencies (up to 400,000 theoretical plates) and shows good reproducibility for migration times (0.5 to 1.0%) and peak heights (2.8 to 7.7%). To demonstrate the rapid analysis of small-volume samples, the adenine-containing compounds present in an oocyte from Xenopus laevis are determined.