Ambulance use affects timely emergency treatment of acute ischaemic stroke.

INTRODUCTION: For acute ischaemic stroke patients, treatment with intravenous tissue plasminogen activator within a 4.5-hour therapeutic window is essential. We aimed to assess the time delays experienced by stroke patients arriving at the emergency department and to compare ambulance users and non-ambulance users. METHODS: We performed a prospective cohort study in a tertiary hospital in Hong Kong. All acute stroke patients attending the emergency department from January to June 2017 were recruited. Patients who were in hospital at the time of stroke onset and those who transferred from other hospitals were excluded. Three phases were compared between ambulance users and non-ambulance users: phase I, between stroke onset and calling for help; phase II, between calling for help and arriving at the emergency department; and phase III, between arriving and receiving medical assessment. RESULTS: Of 102 consecutive patients recruited, 48 (47%) patients arrived at the emergency department by ambulance. The percentage of stroke patients attending emergency department within the therapeutic window was significantly higher for ambulance users than for non-ambulance users (64.6% vs 29.6%; P<0.001). For phases I, II and III, the median times were significantly shorter for ambulance users (77.5, 32 and 8 min, respectively) than for non-ambulance users (720, 44.5 and 15 min, respectively; all P<0.001). CONCLUSION: Transport of patients to the emergency department by ambulance is important for timely and effective stroke treatment.

For t 2 DNA vaccine prevents Forcipomyia taiwana (biting midge) allergy in a mouse model.

BACKGROUND: Forcipomyia taiwana (biting midge) is the most prevalent allergenic biting insect in Taiwan, and 60% of the exposed subjects develop allergic reactions. Subjects with insect allergy frequently limit their outdoor activities to avoid the annoyingly intense itchy allergic reactions, leading to significant worsening of their quality of life. Allergen-specific immunotherapy is the only known therapy that provides long-term host immune tolerance to the allergen, but is time-consuming and cumbersome. This study tested whether the For t 2 DNA vaccine can prevent allergic symptoms in For t 2-sensitized mice. MATERIALS AND METHODS: Two consecutive shots of For t 2 DNA vaccine were given to mice with a 7-day interval before sensitization with recombinant For t 2 proteins, using the two-step sensitization protocol reported previously. RESULTS: The For t 2 DNA vaccine at 50 µg prevented the production of For t 2-specific IgE (P < 0.05), as well as midge allergen-challenge-induced scratch bouts, midge allergen-induced IL-13 and IL-4 production from splenocytes, and inflammatory cell infiltrations in the lesions 48 h after intradermal challenge. CONCLUSIONS: This study is the first to demonstrate that DNA vaccine encoding midge allergen is effective in preventing allergic skin inflammation induced by biting midge. Immunotherapy using For t 2 DNA vaccine can protect mice from being sensitized by midge allergen and may be a promising treatment for biting midge allergy in the future.

Serine protease inhibitor gabexate mesilate attenuates american cockroach-induced bronchial damage and inflammatory cytokine release.

BACKGROUND AND OBJECTIVE: Allergic airway diseases are not only a T,2-mediated chronic airway inflammation, but also a condition of epithelial barrier defects and dysfunction. Allergens with protease activities are known factors that initiate respiratory epithelial damage. Cockroach allergy is the second leading cause of allergic respiratory airway diseases in Taiwan, and cockroach allergens have strong serine protease activity. This study aimed to determine the protective effect of the direct local administration of gabexate mesilate (GM) on American cockroach allergen (CraA)-induced human bronchial epithelial cell inflammation. METHODS: BEAS-2B cells, from the human bronchial epithelial cell line, were stimulated with CraA or co-cultured with different doses of GM. Cellular morphologic changes were observed by microscopy and changes in chemokine mRNA expression and protein levels were determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and ELISA. Effects of specific inhibitors of ERK1/2 (U0126), INK (SP600125), and p38 MAPK (SB203580) on CraA-induced chemokine mRNA expression were also tested by RT-PCR. RESULTS: GM prevented CraA-induced bronchial epithelial cell detachment and morphological changes. It had superior and more extensive suppression effects than specific target MAPK inhibitors in CraA-induced mRNA expression of IL-8, monocyte chemotactic protein (MCP) 1, chemokine (C-C motif) ligand 20, and granulocyte-macrophage colony-stimulating factor from the cells in a dose-dependent manner. CraA-induced IL-8 and MCP-1 protein production from BEAS-2B cells was also attenuated by GM. CONCLUSIONS: The serine protease inhibitor GM has local protective effects against CraA-induced bronchial epithelial inflammation. The development of an inhaled or intranasal protease inhibitor may be a potential strategy for the treatment of allergic airway diseases induced by allergens with protease activities.

Estimating allergenicity of latex gloves using Hev b 1 and hevamine.

BACKGROUND: Latex allergy continues to be an increasingly serious occupational health problem in Taiwan, where it affects approximately 6.8% to 12% of health care workers. Contrasting with reports from western countries, Hev b 1 and hevamine, and not Hev b 3, 5 or 6.02, are the major latex allergens among health care workers in Taiwan. This study aimed at evaluating the allergenicity of 30 brands of commercially available medical latex gloves in Taiwan in 2007. METHODS: Residual Hev b 1 and hevamine from the gloves were measured by inhibition enzyme-linked immunosorbent assay using polyclonal antibodies against purified recombinant Hev b 1 and hevamine. The results were compared to those achieved with quantification of residual total extractable proteins and skin prick testing. RESULTS: The residual extractable protein levels in 30 medical gloves all conformed to United States Food and Drug Administration regulations. All the gloves except one yielded strong skin prick reactions in latex-allergic individuals. The only brand of gloves that consistently produced no skin prick reactions in latex-allergic individuals contained the lowest residual levels of Hev b 1 (0.60 microg/g) and hevamine (0.07 microg/g). CONCLUSIONS: Our results suggest that the measurement of residual extractable total proteins is not sufficient to assess the allergenicity of latex gloves and that Hev b 1 and hevamine may be used as indicator allergens in areas where they are major latex allergens, such as Taiwan.

Responses of midbrain auditory neurons in rats to FM and AM tones presented simultaneously.

Speech and other communication signals contain components of frequency and amplitude modulations (FM, AM) that often occur together. Auditory midbrain (or inferior colliculus, IC) is an important center for coding time-varying features of sounds. It remains unclear how IC neurons respond when FM and AM stimuli are both presented. Here we studied IC neurons in the urethane-anesthetized rats when animals were simultaneously stimulated with FM and AM tones. Of 122 units that were sensitive to the dual stimuli, the responses could be grossly divided into two types: one that resembled the respective responses to FM or AM stimuli presented separately ("simple" sensitivity, 45% of units), and another that appeared markedly different from their respective responses to FM or AM tones ("complex" sensitivity, 55%). These types of combinational sensitivities were further correlated with individual cell's frequency tuning pattern (response area) and with their common response pattern to FM and AM sounds. Results suggested that such combinational sensitivity could reflect local synaptic interactions on IC neurons and that the neural mechanisms could underlie more developed sensitivities to acoustic combinations found at the auditory cortex.

Identification of immunoglobulin E (IgE)-binding epitopes and recombinant IgE reactivities of a latex cross-reacting Indian jujube Ziz m 1 allergen.

Ziz m 1 is a major Indian jujube (Zizyphus mauritiana) allergen involved in latex-fruit syndrome, and cDNA of the allergen has been cloned, sequenced and expressed in yeast by our laboratory previously. In this study, we performed an immunoglobulin E (IgE)-binding epitope analysis of Ziz m 1 using overlapping recombinant fragments. Eight overlapping recombinant fragments were generated from the recombinant Ziz m 1 allergen. The fragments were expressed in Escherichia coli and IgE-binding activities were evaluated by sera of latex-Indian jujube-allergic subjects and normal subjects using immunoblotting. Human allergic sera are not able to recognize fragments consisting of amino acid sequences 26-71, 119-280 and 119-291. However, residues at positions 26-199, 26-105, 26-86, 119-320 and 238-330 were found relevant in the IgE-binding. Our results indicate that (72)NISGHCSDCTFLGEE(86) and (292)VWNRYYDLKTNYSSSIILEYVNSGTKYLP(320) of Ziz m 1 are the sequences required for human IgE binding. Four corresponding peptides, (72)NISGHCSDCTE(86), (292)VWNRYYDLKT(301), (300)KTNYSSSIILEY(311) and (309)LEYVNSGTKYLP(320), were synthesized, and these peptides reacted with 70%, 100%, 70% and 70% of 10 allergic sera tested, as revealed by enzyme-linked immunosorbent assay. Sensitization to (292)VWNRYYDLKT(301) correlated significantly with the presence of allergic symptoms (P < 0.001). These findings will be useful in designing diagnostic and therapeutic approaches, thereby contributing to the development of specific immunotherapy for subjects with latex-fruit syndrome.

A novel haemostatic agent based on self-assembling peptides in the setting of nasal endoscopic surgery, a case series.

INTRODUCTION: Recently, a novel purely synthetic topical haemostatic agent (PuraStat®) has been proposed in surgery based on the self-assembling tendency of some repeating peptide sequences. This transparent, ready to use hydrogel appears suitable for use in FEES with low rates of post-operative re-bleeding and adhesion formation.A first series of 60 patients experiencing endonasal powered turbinoplasty across various hospitals in Sydney using PuraStat® was observed for postoperative re-bleeding and adhesion formation. DISCUSSION: In all 60 patients, no post-operative re-bleeding was observed, while healing went well in absence of adhesion formation. Effective haemostasis with PuraStat® is well documented in other surgical fields, but its use in FEES and adhesion prevention is relatively novel. CONCLUSION: Synthetic self-assembling peptides appear to be indicated in this area. Further studies are needed to confirm their potential for adhesion prevention.

Ca2+-mediated activation of human erythrocyte membrane Ca2+-ATPase.

Ca2+-ATPase of human erythrocyte membranes, after being washed to remove Ca2+ after incubation with the ion, was found to be activated. Stimulation of the ATPase was related neither to fluidity change nor to cytoskeletal degradation of the membranes mediated by Ca2+. Activation of the transport enzyme was also unaffected by detergent treatment of the membrane, but was suppressed when leupeptin was included during incubation of the membranes with Ca2+. Stimulation of the ATPase by a membrane-associated Ca2+-dependent proteinase was thus suggested. Much less 138 kDa Ca2+-ATPase protein could be harvested from a Triton extract of membranes incubated with Ca2+ than without Ca2+. Activity of the activated enzyme could not be further elevated by exogenous calpain, even after treatment of the membranes with glycodeoxycholate. There was also an overlap in the effect of calmodulin and the Ca2+-mediated stimulation of membrane Ca2+-ATPase. While Km(ATP) of the stimulated ATPase remained unchanged, a significant drop in the free-Ca2+ concentration for half-maximal activation of the enzyme was observed.

Sequencing and immunochemical characterization of the American cockroach per a 3 (Cr-PI) isoallergenic variants.

Two additional members of the American cockroach (Periplaneta americana) Per a 3 (Cr-PI) allergen, C13 and C28, were isolated and sequenced. They encoded proteins of 470 and 393 amino acids with two and no potential N-glycosylation sites, respectively. The molecular weights for C13 and C28 cloned proteins are 56,200 and 46,7000, with PI values of 7.06 and 6.54. C13 and C28 display 95.4% identity with several overlapping predicted central antigenic determinants. Both allergens were also found to have a 95% sequence homology with previously cloned C20 and share similar antigenic determinants, as defined by the structural prediction and ELISA analysis. However, the recombinant C13 and C28 allergens showed 26.3 and 94.7% skin reactivities on asthmatic patients while C20 elicited 47.4%. While no sequence similarity was found to other known allergens, these two aromatic amino acid-rich allergens were highly related to insect hemolymph proteins (28.7-36.5%), as with C20 cloned protein. Results suggest that these two are isoallergenic variants of C20. Sequence variations among isoforms, resulting a significant difference in skin reactivities, will be useful in elucidating the allergenic determinants.

Emergence of KPC new variants (KPC-16 and KPC-17) and ongoing outbreak in southern Taiwan.

We first describe two novel variants of blaKPC, blaKPC-16 and blaKPC-17, which were identified in three Klebsiella pneumoniae isolates from a patient in Taiwan. KPC-16 and KPC-17 differed from KPC-2 by two (P202S and F207L) and a single (F207L) amino acid substitutions, respectively. All three isolates with identical pulsotype belonged to sequence type 11. The MICs of the three isolates for colistin and tigecycline were 0.5 µg/mL and 2 µg/mL, respectively. Moreover, an outbreak of at least 39 blaKPC-17-containing K. pneumoniae isolates is ongoing in southern Taiwan in 2014. Physicians should know that blaKPC-17-containing isolates can substantially threaten public health.

A comparison of CNS angiotensin II binding in the hypothalamus-thalamus-septum-midbrain region of developing spontaneously hypertensive and normotensive rats.

Specific angiotensin II (ANG II) binding was studied in brain homogenates from the hypothalamus-thalamus-septum-midbrain (HTSM) region of age-matched 4-, 8-, 12- and 16-week spontaneously hypertensive rats (SHR) and their normotensive controls, Wistar-Kyoto (WKY) rats, using 125I-angiotensin II. Scatchard analysis revealed that the dissociation constants (Kd) ranged from 0.36 to 0.73 nmol/l, although these values were not significantly different at any given age period between the SHR and WKY rats. In contrast, a statistically significant increase in ANG II receptor binding was seen between the SHR and WKY rat at 4 weeks of age. However, this difference was not observed at older age periods. Furthermore, both the SHR and WKY rat showed a decrease in ANG II receptor levels during development, with the most marked reductions occurring between 12 and 16 weeks of age for both strains. These findings suggest that ANG II receptors in the HTSM region of both the SHR and WKY rat are down-regulated during development, that receptor loss is more significant in the SHR than in its normotensive control and that binding capacity differences between the two strains are only seen before the onset of measureable increases in the arterial pressure of the SHR. We conclude that there is a significant difference in the ANG II binding capacity during the development of hypertension in the SHR as compared with the WKY rat and therefore it may play a role in the pathogenesis of this disorder.

Human adult-onset lactase decline: an update.

Human adult-onset lactase decline is a biologic feature characteristic of the maturing intestine in the majority of the world's population. The digestion and absorption of lactose, the major carbohydrate in milk and also the main substrate for lactase, is often variable, a consequence of lactase levels, gastric emptying rate, and colonic salvage. Although commercially available "lactase" products alleviate symptoms in many lactose-intolerant people, a greater understanding of this variability in lactose tolerance could lead to interventions that reduce the rate of gastric emptying and/or increase the proliferation of lactose-metabolizing bacteria in the colon, leading to more efficient lactose utilization. Adult-onset lactase decline appears to be a risk factor for developing osteoporosis, owing to avoidance of dairy products or interference of undigested lactose with calcium absorption. Elderly with both adult-onset lactase decline and atrophic gastritis or those undergoing anti-ulcer treatment may have an increased risk of low calcium absorption owing to the lack of gastric acid that facilitates calcium uptake. Thus, lactose-intolerant elders should monitor their calcium nutrition status carefully.

Effects of guanyl nucleotides and ions on kappa opioid binding.

Displacement curve analyses demonstrated that GTP and its nonhydrolyzable analog, GPP(NH)P, inhibited the binding of [3H]dihydromorphine (mu agonist), [3H]D-Ala2-D-Leu5-enkephalin (delta agonist), and [3H]ethylketocyclazocine (general agonist) but not [3H]diprenorphine (general antagonist). Using a paradigm to block mu and delta sites with specific cold ligands, [3H]ethylketocyclazocine labeled kappa sites which were less GTP sensitive than sites labeled by mu and delta agonists. Further, Na+ and Mg++, important in inhibitory adenylate cyclase systems, also decreased both unblocked and mu-/delta-blocked [3H]ethylketocyclazocine binding. Scatchard analyses revealed that the inhibitory effects of GTP result in a decrease in affinity without a significant change in binding capacity, and that the kappa component of [3H]ethylketocyclazocine binding was less sensitive to the effects of GTP than binding sites labeled by mu or delta agonists. In comparison to the effects of GTP, Na+ decreased binding affinity but increased the binding capacity of the kappa component. These data also suggest that the inhibitory effects of Na+ and GTP on binding affinity are not additive. Association and dissociation plots revealed that although both components of binding may be involved in these affinity changes, the dissociation rate represents the more significant factor. These data suggest that [3H]ethylketocyclazocine binding to kappa sites is GTP and Na+ sensitive. However, it should be noted that [3H]ethylketocyclazocine binding to kappa sites is less sensitive to GTP than its binding to other opiate sites, and that this kappa binding is differentially affected by Na+. The significance of these characteristics with regard to the effect of kappa opiates on adenylate cyclase activity remains to be determined.

Detection of human papillomavirus types in cervical adenocarcinoma by the polymerase chain reaction.

OBJECTIVE: To determine the human papillomavirus (HPV) types in cervical adenocarcinoma of patients from Taiwan. METHODS: DNA was extracted from fixed tissues and polymerase chain reaction was performed in conjunction with a unique probe, pRSA I, allowing simultaneous detection of HPV types 6, 11, 16, 18, 31 and 33 from amplified HPV DNAs after endonuclease, RsaI, digestion. RESULTS: Of 69 tissues examined, 31.9% (22/69) were found to contain HPV DNA. Among 22 HPV-positive specimens, no HPV types 6, 11, 31 and 33 were detected. On the other hand, HPV 16 and HPV 18 were found in 11 (15.9%) and 10 (14.5%) of HPV-positive specimens, respectively. One specimen (1.5%) was found to contain both HPV 16 and 18 DNAs. CONCLUSIONS: Our findings support that HPV 18, along with HPV 16, may play a certain role in the adenocarcinoma pathogenesis of the uterine cervix.

Etiology and treatment of post-cesarean-section endometritis after cephalosporin prophylaxis.

To describe the microbiologic etiology of post-cesarean endometritis developing after perioperative cephalosporin prophylaxis, endometrial samples were obtained from 27 women with a triple-lumen catheter. The women were assigned in a double-blind, randomized fashion to receive either ticarcillin/clavulanate, 3.1 g, or cefoxitin, 2 g, administered every six hours, until the clinical signs of infection resolved. A total of 149 microorganisms (84 facultative and 65 obligate anaerobes) were recovered from 26 women, for a mean of 5.5 isolates per specimen. One endometrial specimen was sterile. Bacteroides and Peptostreptococcus species were the most frequent isolates, followed by Gardnerella vaginalis, Enterococcus, facultative gram-negative rods and Mycoplasma hominis. Each of the isolates was tested for beta-lactamase activity. At least one beta-lactamase-producing isolate was recovered from 56% of the specimens. Susceptibility testing of endometrial isolates demonstrated that 96% of 118 potential pathogens (Gardnerella, Bacteroides, Peptostreptococcus, enterococci and streptococci) were susceptible to ticarcillin/clavulanate. By comparison, 86% of these isolates were susceptible to cefoxitin in vitro. Women who were treated with ticarcillin/clavulanate were less likely to have a temperature greater than 38 degrees C for two or more days (8% vs. 57%, P = .01). Also, there was a trend toward a decreased duration of uterine tenderness in the ticarcillin/clavulanate group, but it did not attain statistical significance (60% vs. 86%, P = .4). However, the overall clinical success rate with these single-agent treatments was not different for the two groups (77% vs. 79%, P = 1.0).(ABSTRACT TRUNCATED AT 250 WORDS)

Tarsal tunnel syndrome caused by talocalcaneal coalition.

Tarsal tunnel syndrome caused by talocalcaneal coalition is uncommon. We presented the ultrasonography (US) and magnetic resonance imaging findings of this disease. This is, to our knowledge, the first case report describing the US findings in tarsal tunnel syndrome caused by talocalcaneal coalition.

The endocrine mechanism of sex reversal in the protandrous black porgy, Acanthopagrus schlegeli: a review.

Black porgy, Acanthopagrus schlegeli Bleeker, a marine protandrous hermaphrodite, is a functional male for the first 2 years of life but begin to sexually reverse to female after the third year. This sex pattern provides a very good model to study the mechanism of sex reversal in fish. The gonad at 5 month of age consisted of testicular tissue with few primary oocytes at 5 month of age. The ovarian tissue became dominant at 18 months of age during the non-spawning season. Testicular and ovarian tissues were separated by connective tissue. Plasma estradiol-17 beta(E2), vitellogenin and 11-ketotestosterone (11-KT) profiles in males were significantly different from those in the 3-year-old reversing females. Peak levels of plasma E2 in the reversing females occurred during the early prespawning season (in October). Lower levels of plasma E2 were, however, observed in the males. Plasma 11-KT levels significant decreased but no changes of plasma testosterone were detected in the reversing females. Exogenous E2 suppressed the testicular development but induced the gonadal aromatase activity, ovarian development and sex reversal in 2-year-old black porgy. Exogenous T and LHRH analog did not have effects on the sex reversal. Higher concentrations of pituitary GtH II and mRNA of GtH II-beta subunit were detected in the reversed females. These data suggested that E2 and gonadal aromatase closely associated to the occurrence of sex reversal. A working model of the sex reversal in black porgy is proposed.