Association of Protein Expression p53 Mutants with Regional Lymph Gland Status on type III Carcinoma Nasofaring Patients.
Nasopharyngeal carcinoma (NPC) is a malignant disease originating from the nasopharyngeal epithelial cells. The molecular mechanism of cancer occurrence is a change in the oncogene and tumor suppressor genes. One of the tumor suppressor genes that mutate in cancer cells is the mutant p53 gene. One of nasopharyngeal carcinoma progression is determined by the status of regional lymph gland. The enormous regional lymph node has a poor prognosis. To analyze the expression of the mutant p53 protein in Nasopharyngeal carcinoma (NPC) that correlated with regional lymph gland status (N) as a clinical manifestation. Expression of mutant p53 protein from NPC tissue paraffin block with immunohistochemical cracking technique was using monoclonal rabbit Anti Human p53 clone 318-6-11 (Dako, North America, Inc., 6392 Via Real Carpinteria, CA 93013 USA), microscope light binoculars was assessed visually by an Anatomical Pathology Consultant. Positive expression of p53 mutants was obtained 57.58% from all the sample in N0 by 0 subjects, N1 was 6 subjects, N2 was 7 subjects, and N3 was 7 subjects. The results of Mann-Whitney U test was p = 0.706, then there was no significant (p > 0.05) correlation between positive expression of p53 protein in type III WHO NPC with the regional lymph gland were N0, N1, N2, and N3. There was no significant between expression protein p53 mutants' regional and lymph gland in type III WHO NPC.


Synthesis and antileishmanial activities of 4,5-di-substituted acridines as compared to their 4-mono-substituted homologues.
Newly synthesized 4,5-di-substituted acridines were assessed for in vitro antileishmanial activities as compared to those of their 4-mono-substituted homologues. Mono-substituted acridines exhibited a weak specificity for Leishmania parasites. Di-substituted acridines, on the contrary, displayed interesting amastigote-specific activities through a mechanism of action that might not involve intercalation to DNA. This antileishmanial property, associated with a low antiproliferative activity towards human cells, led to the identification of a new class of promising acridine derivatives such as 4,5-bis(hydroxymethyl)acridine with a nonclassical mechanism of action based on the inhibition of Leishmania internalization within macrophages. In the meantime, the effects of experimental lighting on the biological properties of acridines were assessed: experimental lighting did not significantly improve the antileishmanial activity of the compounds since it produced a greater toxicity against human cells.


Effects of G-CSF on telomere lengths in PBMCs from human immunodeficiency virus-infected patients: results from a randomized, placebo-controlled trial.
Telomeres are unique terminal chromosomal structures, the length of which has been shown to decrease with cell division in vitro and with increased age in vivo for human somatic cells. In human immunodeficiency virus (HIV)-1 infection, decrease of telomere length is primarily found in CD8+ T cells, and not in CD4+ T cells. In this double-blind placebo-controlled study, we investigated the effect of granulocyte colony stimulating factor (G-CSF) treatment combined with highly active antiretroviral therapy (HAART) on mean telomere length in peripheral blood mononuclear cells (PBMC). The terminal restriction fragment (TRF) length showed no changes during G-CSF treatment although the number of lymphocytes increased significantly. The mean TRF length correlated positively (R = 0.552, P = 0.009) and negatively (R = -0.503, P = 0.02) to the proportion of CD4+ memory and naïve cells, respectively. Our data suggest that during G-CSF treatment lymphocytes are recruited by a combination of central and peripheral proliferation.


Gender and nonmedical prescription opioid use and DSM-5 nonmedical prescription opioid use disorder: Results from the National Epidemiologic Survey on Alcohol and Related Conditions - III.
Little is known about sex-specific risk for nonmedical prescription opioid use (NMPOU) and DSM-5 nonmedical prescription opioid use disorder (NMPOUD). The objective of the present study was to present prevalence, correlates, psychiatric comorbidity, treatment and disability of NMPOU and DSM-5 NMPOUD among men and women. Nationally representative sample of the U.S. Prevalences of 12-month and lifetime NMPOU were greater among men (4.4%, 13.0%) than women (3.9%, 9.8%), while corresponding rates of DSM-5 NMPOUD did not differ between men (0.9%, 2.2%) and women (0.9%, 1.9%). Regardless of time frame and sex, NMPOU and NMPOUD generally decreased with age and were lower among Blacks, Asians/Pacific Islanders and Hispanics, and respondents with lower socioeconomic status. Among men with NMPOU, rates were lower among respondents in the Northeast and South and among those previously married (lifetime). Across time frames and gender, NMPOU and NMPOUD were generally associated with other substance use disorders, posttraumatic stress and borderline, schizotypal and antisocial personality disorders, but associated with major depressive disorder, persistent depression and bipolar I disorder only among men. Disability increased with NMPOU frequency and NMPOUD severity. Only 7.6% and 8.2% of men and women with NMPOU ever received treatment, while 26.8% and 31.1% ever received treatment for NMPOUD. NMPOU and NMPOUD are highly disabling, associated with a broad array of sex-specific and shared correlates and comorbidities and largely go untreated in the U.S. Valid assessment tools are needed that include gender as a stratification variable to identify NMPOU and NMPOUD.


Review article laser-induced hyperthermia on graphene oxide composites.
Hyperthermia-based therapies have shown great potential for clinical applications such as for the antitumor and antipathogenic activities. Within all strategies, the so-called photothermal therapy proposes to induce the hyperthermia by the remote laser radiation on a photothermal conversion agent, in contact with the target tissue. This paper reviews the most relevant in vitro and in vivo studies focused on NIR laser-induced hyperthermia due to photoexcitation of graphene oxide (GO) and reduced graphene oxide (rGO). Relevant parameters such as the amount of GO/rGO, the influence of the laser wavelength and power density are considered. Moreover, the required temperature and exposure time for each antitumor/antipathogenic case are collected and unified in a thermal dose parameter: the CEM43. The calculated CEM43 thermal doses revealed a great variability for the same type of tumor/strain. In order to detect potential tendencies, the values were classified into four ranges, varying from CEM43 < 60 min to CEM43 ≥ 1 year. Thus, a preference for moderate thermal doses of CEM43 < 1 year was detected in antitumor activity, with temperatures ≤ 50 °C and exposure time ≤ 15 min. In case of the antipathogenic studies, the most used thermal dose was higher, CEM43 ≥ 1 year, with ablative hyperthermia (> 60ºC). The ability of GO/rGO as effective photothermal conversion agents to promote a controlled hyperthermia is proven. The variability found for the CEM43 thermal doses on the reviewed studies reveals the potentiality to evaluate, for each application, the use of lower temperatures, by modulating time and/or repetitions in the doses.


Novel imidazo[1,2-a]pyrazine derivatives as potent reversible inhibitors of the gastric H+/K+-ATPase.
A series of novel 6-substituted imidazo[1,2-a]pyrazines were synthesized via palladium catalyzed amino- or alkoxycarbonylation as key step. The anti-secretory activity of these compounds has been assessed in a binding assay against H(+)/K(+)-ATPase from hog gastric mucosa. Some of the compounds proved to be potent inhibitors of the gastric acid pump.


When sad groups expect to meet again: interactive affective sharing and future interaction expectation as determinants of work groups' analytical and creative task performance.
The present study examines the moderating role of future interaction expectation in the relationship between affective sharing and work groups' task performance. We argue that group affect, a group defining characteristic, becomes more salient to its members when it is interactively shared, and that the anticipation of future interaction may strengthen the effects of group defining characteristics on subsequent group member behaviour. As a consequence, interactive sharing (vs. non-interactive sharing) of negative affect is more likely to influence work group outcomes when group members expect to meet again. Results from a laboratory experiment with 66 three-person work groups indeed show that interactively shared (vs. non-interactively shared) negative affect facilitated work groups' analytical task performance, whereas it inhibited performance on a creative fluency task when groups have expectations of future interaction and not when they do not have such expectations. The discussion focuses on how these results add to theory on group affect and contribute to insights in the effects of future interaction expectation.


Contact allergy to corticosteroids: Is the European baseline series sufficient?
Patients are consecutively screened for contact allergy to corticosteroids with budesonide and tixocortol-21-pivalate in the European baseline series. Centres using TRUE Test also include hydrocortisone-17-butyrate. A supplementary corticosteroid patch test series is used in case of suspicion of corticosteroid contact allergy or when a marker of corticosteroid contact allergy is positive. The aims were to evaluate (1) the efficacy of corticosteroids in the TRUE Test and (2) co-sensitization patterns. This retrospective study analysed patients patch tested with TRUE Test corticosteroids plus supplementary corticosteroid series in the period 2006-2020 at the Department of Dermatology and Allergy Centre, Odense University Hospital. Of 1852 patients tested, 119 were sensitised to TRUE Test corticosteroids and supplementary testing found additional reactions to other corticosteroids in 19 of 119 patients. TRUE Test corticosteroids gave more positive and stronger reactions compared to allergens in petrolatum/ethanol. Fourteen percent of sensitised patients were co-sensitised to multiple corticosteroid groups. Baeck group 3 corticosteroids accounted for 9 of 16 patients not identified by TRUE Test. Budesonide, hydrocortisone-17-butyrate, and tixocortol-21-pivalate in combination are sensitive corticosteroid markers. In case of clinical suspicion of corticosteroid contact allergy, patch testing with supplementary corticosteroids is highly recommended.


Reaction of small heat-shock proteins to different kinds of cellular stress in cultured rat hippocampal neurons.
Upregulation of small heat-shock proteins (sHsps) in response to cellular stress is one mechanism to increase cell viability.We previously described that cultured rat hippocampal neurons express five of the 11 family members but only upregulate two of them (HspB1 and HspB5) at the protein level after heat stress. Since neurons have to cope with many other pathological conditions, we investigated in this study the expression of all five expressed sHsps on mRNA and protein level after sublethal sodium arsenite and oxidative and hyperosmotic stress. Under all three conditions, HspB1, HspB5, HspB6, and HspB8 but not HspB11 were consistently upregulated but showed differences in the time course of upregulation. The increase of sHsps always occurred earlier on mRNA level compared with protein levels. We conclude from our data that these four upregulated sHsps (HspB1, HspB5, HspB6, HspB8) act together in different proportions in the protection of neurons from various stress conditions.


[Prevalence of childhood cerebral palsy in six provinces in China].
To investigate the prevalence of childhood cerebral palsy (CP) in China. A cross sectional survey was carried out among 1047327 children aged 1 approximately 6 years old in six provinces of China during May 1997 approximately December 1998. The crude prevalence of CP for children aged 1 approximately 6 years old was 1.92 per thousand; in addition, prevalences of CP were also reported for factors such as sex, age, ethic group, birth weight, gestational age, and area. Prevalence of CP in China is at the low range among the world, which may be due to low prevalence and low survival rate of premature newborn in China. The prevalence of cp is associated with areas, sex, age, ethnic group, birth weight, and gestational age.


The kidney and aldosterone in acclimatization at altitude.
Five climbers were studied during an ascent of a 7500 m mountain in the Hindu Kush. Representiative samples from 24-hour urines were preserved from five subjects at 5400 m and 6600 m and from two at 7000 m. Sodium and cortisol excretion was normal, but that of potassium and especially aldosterone was low, especially in two climbers who had episodes of peripheral oedema. Hypoaldosteronism is a feature of high altitude acclimatization but its cause and significance have still to be elucidated.


Cytochrome bd biosynthesis in Bacillus subtilis: characterization of the cydABCD operon.
Under aerobic conditions Bacillus subtilis utilizes a branched electron transport chain comprising various cytochromes and terminal oxidases. At present there is evidence for three types of terminal oxidases in B. subtilis: a caa3-, an aa3-, and a bd-type oxidase. We report here the cloning of the structural genes (cydA and cydB) encoding the cytochrome bd complex. Downstream of the structural genes, cydC and cydD are located. These genes encode proteins showing similarity to bacterial ATP-binding cassette (ABC)-type transporters. Analysis of isolated cell membranes showed that inactivation of cydA or deletion of cydABCD resulted in the loss of spectral features associated with cytochrome bd. Gene disruption experiments and complementation analysis showed that the cydC and cydD gene products are required for the expression of a functional cytochrome bd complex. Disruption of the cyd genes had no apparent effect on the growth of cells in broth or defined media. The expression of the cydABCD operon was investigated by Northern blot analysis and by transcriptional and translational cyd-lacZ fusions. Northern blot analysis confirmed that cydABCD is transcribed as a polycistronic message. The operon was found to be expressed maximally under conditions of low oxygen tension.


Nanogold probe enhanced Surface Plasmon Resonance immunosensor for improved detection of antibiotic residues.
An exhaustive study is reported on the effect that antibody nanogold probes produce on the performance of a Surface Plasmon Resonance (SPR) immunosensor. The paper studies the improvement that different nanogold probes prepared at different antibody:gold nanoparticle (IgG:AuNP) ratios and AuNP sizes produce on the maximum signal and detectability of a simple SPR immunosensor developed to analyze fluoroquinolone (FQ) antibiotic residues (SPReeta system). The investigation compares the features of sensor enhanced formats using both, secondary and primary nanogold probes (anti-IgG and IgG coupled to AuNP, on double and single-antibody immunochemical assay steps, respectively), in respect to the unenhanced format. For this purpose, a reproducible bioconjugation procedure for preparing gold biohybrid nanoparticles has been established, involving the formation of a mixed self-assembled monolayer (m-SAM) with PEGylated cross-linkers around the AuNP followed by the covalent attachment of the antibodies. The procedure allows controlling the IgG:AuNP ratio of the nanogold probes on a reproducible manner and the functionalized NPs have been found to be stable during assay and storage. Both formats, using secondary and primary nanogold probes, are excellent strategies to improve immunosensor detectability. Thus, using anti-IgG-AuNP, the detectability could be improved by a factor of 14 (LOD 0.07±0.01 μg L(-1) vs. 0.98±0.38 μg L(-1)) reducing at the same time the amount of primary antibody used (30,000 vs. 1000 dilution factor). Likewise, the format using IgG-AuNP also allows improving detectability (LOD 0.11±0.01 μg L(-1)), but reducing the number of needed steps.


Substance P neuronal organization in the median region of the interpeduncular nucleus of the cat: an electron microscopic analysis.
The localization of substance P-like immunoreactivity in the interpeduncular nucleus using the peroxidase-antiperoxidase technique, revealed that the median region of the interpeduncular nucleus was one area rich in substance P-like immunoreactive processes. The ultrastructural characteristics of these substance P-like immunoreactive processes and their organization within the middle zone of the median region of the interpeduncular nucleus was studied. Substance P-like immunoreactivity was found in the perikaryon of small neurons, and in proximal and small dendrites. The substance P-like immunoreactive cell bodies and proximal dendrites receive a variety of unlabeled synaptic terminals. The immunoreactive small dendrites usually formed the central component of a "rosette"-like formation with unlabeled terminals. A few immunoreactive small unmyelinated axons and boutons were also present in the neuropil. The substance P-like immunoreactive boutons contained mainly small round vesicles with some large dense-core vesicles. These substance P-like immunoreactive boutons were presynaptic to unlabeled dendritic profiles, and frequently to substance P-like immunoreactive dendritic profiles. They were also seen in apposition to unlabeled boutons. Substance P-like immunoreactive boutons were not found to synapse with the crest-like dendritic processes in this part of the interpeduncular nucleus. It is suggested on the basis of morphological features, that some of the unlabeled terminals synapsing on substance P-like immunoreactive dendrities, may be cholinergic in nature.


[Arteriovenous fistula of the renal pedicle following nephrectomy responsible for heart failure. Report of 2 cases].
The authors report two cases of acquired post-nephrectomy arteriovenous fistula. The large shunt was responsible for cardiac failure. The first case was treated by percutaneous embolisation and the second by reoperation. The clinical results were excellent. These two cases illustrate the value of different imaging techniques and therapeutic approaches: direct surgery or embolisation.


Persistence of high intestinal lactase activity in Pakistan.
In order to determine the incidence of persistence of high intestinal lactase activity (PLA) in a Pakistani human population, 53 probands belonging to the Punjabi ethnic group were examined using a lactose tolerance test. The incidence of PLA in the sample was found to be 55%, which has been compared with the incidence in other ethnic groups racially or geographically related to the Pakistani population. Various possible explanations for the fairly high incidence of PLA in the Punjabi ethnic group have been discussed.


Multiple myeloma: Detection of free monoclonal light chains by modified immunofixation electrophoresis with antisera against free light chains.
Neoplastic monoclonal gammopathies are frequently associated with production of excess free monoclonal light chains. A sensitive method for detecting free monoclonal light chains in serum could provide a marker for residual/minimal residual disease and as an adjunct to serum protein electrophoresis to serve as a screening method for monoclonal gammopathies. Conventional serum immunofixation electrophoresis was modified by applying undiluted serum samples, and staining for serum free light chains with antisera specific to free light chains. Washing/blotting of gels was enhanced to remove residual proteins that did not bind to the antibodies including intact monoclonal immunoglobulins. Results from this modified immunofixation electrophoresis were compared with results from commercially available MASS-FIX/MALDI assay. Monoclonal free kappa light chains could be detected to a concentration of about 1.78 mg/L and monoclonal free lambda light chains to a concentration of about 1.15 mg/L without the need for special equipment. Comparison of these thresholds with parallel results from a commercially available MASS-FIX/MALDI assay revealed the modified electrophoretic method to be more sensitive in the context of free monoclonal light chains. Modified serum immunofixation electrophoresis has been shown to detect low levels of monoclonal free light chains at a sensitivity suitable for the method to be used in detecting minimal residual disease, and potentially in a screening assay for monoclonal gammopathies. The disparity in the results with commercially available MASS-FIX/MALDI assay is postulated to be due to limited repertoire of reactivity of nanobodies of camelid origin.


Clinical and Urodynamic Predictors for Failure of Medical Management with Alpha Blockers in PBNO: A Retrospective Cohort Analysis.
To evaluate the clinical and urodynamic variables that may predict the failure of alpha-blockers in primary bladder neck obstruction (PBNO) patients. Alpha-blockers are useful as a treatment option in patients with PBNO. Nonresponders need to undergo bladder neck incision (BNI). Little is known about the predictive factors determining the success of treatment. This was a retrospective study, spanning over a period of 8 years. PBNO was diagnosed in the presence of a bladder outlet obstruction index (BOOI) >40 with video-urodynamic evidence of obstruction at the bladder neck. The patients were initially managed with alpha-blockers (alfuzosin and tamsulosin) for 3-6 months, and BNI contemplated when pharmacotherapy failed. The patients with upper tract changes managed with upfront BNI or clean intermittent catheterization were excluded. The data for the international prostate symptom score (IPSS), uroflowmetry, urodynamic studies, and ultrasonography of pre and post-treatment periods were reviewed. Treatment outcomes were defined as complete response (>50% improvement in Qmax and IPSS score) and partial response (30%-50% improvement in Qmax and IPSS score) at 3 or 6 months. Ninety-nine patients were analyzed. 21 patients underwent BNI for the failure of medical management and 31 for recurrence of symptoms at a mean follow-up of 18.8 ± 3.5 months (12-70 months). Independent predictors of failure of pharmacotherapy with alpha-blockers were age (P = .021), Pdet@Qmax (P = .015), and BOOI (P = .019). Alpha-blockers are more likely to fail in PBNO in younger patients generating higher voiding pressures and BOOI > 60.


In vitro tests to evaluate potential biological activity in natural substances.
The biological activity of polysaccharides from the mycelia of 40 Basidiomycetes was studied using an uncommon toxicity test technique, the planaria bioassay, and the better known potato disk bioassay. The results showed the utility of this duo of 'in vitro' tests as a preliminary screening of the toxicity of substances that are present in aqueous fungal extracts.


Immunogenicity and safety profiles of genetic vaccines against human Her-2/neu in cynomolgus monkeys.
Her-2/neu is a well-characterized tumor-associated antigen, the overexpression of which in human carcinomas correlates with a poor prognosis. Here, we evaluated Her-2/neu-specific humoral and cellular immune responses in immunized monkeys after immunization with nonreplicating adenovirus (AdHM) expressing the extracellular and transmembrane domain of human Her-2/neu (HM) and/or naked DNA vaccine (pHM-hGM-CSF) expressing human granulocyte-macrophage colony-stimulating factor together with HM. Priming of monkeys with AdHM generated Her-2/neu-specific long-lasting antibody production. Furthermore, these Her-2/neu-specific antibodies produced by AdHM immunization, some of which shared epitope specificity with Herceptin, were able to induce antibody-dependent cellular cytotoxicity against Her-2-expressing target cells. Cellular immune responses were elicited in all monkeys immunized with Her-2/neu-expressing vaccine; interferon-gamma was secreted when these splenocytes were restimulated with Her-2/neu-expressing autologous cells, and immunization with AdHM induced Her-2/neu-specific lymphoproliferative responses. Further, immunization with pHM-hGM-CSF before AdHM immunization noticeably enhanced cytotoxic T-lymphocyte activity. In addition, we observed no abnormalities that would indicate that the genetic vaccines had toxic effects in the immunized monkeys. Thus, we can conclude that our genetic vaccines efficiently elicited Her-2/neu-specific humoral and cellular immune responses without causing severe adverse effects in nonhuman primates and that as such they warrant further clinical investigation.


A Canadian urban early pregnancy assessment clinic: a review of the first year of operation.
To review the services provided by an early pregnancy assessment clinic (EPAC) in Vancouver, British Columbia, and to assess the characteristics of the women accessing their services and their satisfaction with the care provided. The EPAC at BC Women's Hospital and Health Centre was opened in September 2007 to offer assessment of early pregnancy complications and management of early pregnancy loss. We conducted a review of the records of all patients who accessed the clinic in its first year of operation to review the origin of referrals, patient assessments, diagnoses, treatment, complications, and follow-up. Additionally, we distributed satisfaction surveys to patients. A total of 411 patients were referred to the EPAC in its first year of operation. Most referrals were from family physicians (38%), and the primary reason for referral was most commonly cramping and/or spotting (57%). The mean wait time for patients to be seen was 2.02 working days, and the majority of women (91%) were given a diagnosis at their first clinic visit. After assessment, 48% of women had manual vacuum aspiration or dilatation and curettage arranged, 21% had expectant management, and 17% required no treatment, because of complete miscarriage. While 94% of patients with non-viable pregnancies were treated at EPAC, 6% were referred to other health care agencies. In the satisfaction surveys completed, comments about the EPAC were largely positive and primarily focused on the emotional support and level of care received. In its first year of operation, the EPAC was well used and provided efficient and compassionate care. Patients reported high satisfaction with the quality of care received from staff, and showed particular appreciation for the emotional care received.


Patient characteristics associated with different types of prison TB: an epidemiological analysis of 921 TB cases diagnosed at an Ethiopian prison.
Despite incarcerated population being at an increased risk of tuberculosis (TB) and serving as a potential source of TB transmission for the general population, prison TB remains understudied. Given its adverse impact on progress towards TB elimination, World Health Organization (WHO) has identified prison TB research as a top priority to guide TB treatment/control interventions. We retrospectively analyzed 921 notified TB cases that were diagnosed at Kality Federal Prison, Ethiopia during 2009-2017. To assess trends of microbiologically confirmed pulmonary TB (PTB), extra-pulmonary TB (EPTB), and TB-HIV co-infection, an ecological analysis of aggregated cases was used to report trends over time. Additionally, we used multivariable log binomial regression to identify patient characteristics associated with microbiologically confirmed PTB, EPTB, and TB-HIV co-infection. Microbiologically confirmed PTB proportion increased over time. Young age was identified as an important risk factor for EPTB (adjusted prevalence ratio [aPR] = 1.74, 95% CI 0.97, 3.13) while HIV coinfection was negatively associated with EPTB (aPR = 0.73, 95% CI 0.55, 0.97). While previous TB history was associated with a lower likelihood of EPTB (aPR = 0.42, 95% CI 0.25, 0.70), it was associated with an increased risk of TB-HIV coinfection (aPR = 1.37, 95% CI 1.10, 1.71). Clinically diagnosed PTB patients were more likely to have TB-HIV coinfection compared to microbiologically confirmed PTB patients (aPR = 1.32, 95% CI 1.02, 1.72). Increasing proportion of microbiologically confirmed PTB may suggest delayed access to treatment, severe disease and increased risk of intramural transmission. Associations with clinical/demographic factors varied for different types of TB and were not always consistent with what has been previously reported for the general population, necessitating the need to refocus prison TB control/treatment strategies based on context specific epidemiological factors.


Two overlapping two-component systems in Xanthomonas oryzae pv. oryzae contribute to full fitness in rice by regulating virulence factors expression.
Two-component signal transduction systems (TCSs) are widely used by bacteria to adapt to the environment. In the present study, StoS (stress tolerance-related oxygen sensor) and SreKRS (salt response kinase, regulator, and sensor) were found to positively regulate extracellular polysaccharide (EPS) production and swarming in the rice pathogen Xanthomonas oryzae pv. oryzae (Xoo). Surprisingly, the absence of stoS or sreKRS did not attenuate virulence. To better understand the intrinsic functions of StoS and SreKRS, quantitative proteomics isobaric tags for relative and absolute quantitation (iTRAQ) was employed. Consistent with stoS and sreK mutants exhibiting a similar phenotype, the signalling circuits of StoS and SreKRS overlapped. Carbohydrate metabolism proteins and chemotaxis proteins, which could be responsible for EPS and swarming regulation, respectively, were reprogrammed in stoS and sreK mutants. Moreover, StoS and SreKRS demonstrated moderate expression of the major virulence factor, hypersensitive response and pathogenicity (Hrp) proteins through the HrpG-HrpX circuit. Most importantly, Xoo equipped with StoS and SreKRS outcompetes strains without StoS or SreKRS in co-infected rice and grows outside the host. Therefore, we propose that StoS and SreKRS adopt a novel strategy involving the moderation of Hrp protein expression and the promotion of EPS and motility to adapt to the environment.


Blood Meal Analysis and Molecular Detection of Leishmania DNA in Wild-Caught Sand Flies in Leishmaniasis Endemic Areas of Turkey and Northern Cyprus.
Phlebotomine sand flies (Diptera: Psychodidae) are known as the vector of diseases such as leishmaniasis, bartonellosis and viral diseases. The aim of this study is to detect the host feeding pattern of sand flies in the endemic areas for leishmaniasis in Turkey (Antalya, Kayseri) and Northern Cyprus (TRNC) as well as the presence of Leishmania DNA in the specimens. One-hundred seventy-six blood-fed sand fly specimens were examined for blood meal analysis. A SYBR Green-PCR assay was performed with specific forward primers for each host and a universal reverse primer. Primers of human and goat were used together in multiplex PCR while goat and cow were studied separately. ITS-1 qPCR assay was also performed on both blood-fed and non-blood-fed females to detect Leishmania parasites. Blood sources could be detected in 69 out of 176 blood-fed sand fly specimens. The results of blood meal analysis showed that specimens were fed mostly on cows (22.2%) followed by humans (5.7%), goats (2.8%) and dogs (0.6%). Multiple feeding patterns were also detected as human + cow (3.4%), cow + goat (2.8%) and human + goat (1.7%). Five of the blood-fed specimens were Leishmania spp. positive: P. major s.l. (n = 1), P. tobbi (n = 2) were L. tropica positive from Antalya, P. simici was positive for L. infantum from Kayseri and P. papatasi (n = 1) was positive for L. major from Cyprus. Leishmania infection rates were determined as 3.79%, 1.69% and 2.63% among the blood-fed sand fly specimens in Antalya, Kayseri and TRNC, respectively. The SYBR-Green-based multiplex PCR assay is a cost-effective and promising tool for blood meal identification of wild-caught sand flies as well as other blood-sucking arthropods. Feeding patterns of important vector species detected in the present study show the high risk in these endemic areas. As a next step, to identify the blood source in a shorter time and to make the test more sensitive, development of this assay to probe-based and multiplex PCR will be also planned.


Multicenter Evaluation of Xpert Carba-R Assay for Detection and Identification of the Carbapenemase Genes in Rectal Swabs and Clinical Isolates.
Rapid detection of carbapenemase-producing organisms is clinically desirable for hospital infection control and antibiotic stewardship. In this multicenter study, the Xpert Carba-R assay was evaluated for detection of the five carbapenemase genes (blaKPC, blaNDM, blaIMP, blaOXA-48, and blaVIM) in 2404 nonduplicate rectal swabs of admitted inpatients and 521 Gram-negative isolates from four tertiary hospitals in China, compared with the reference growth-based method with DNA sequence analysis of colonies. All suspected false-positive results in rectal swabs were resolved by supplementary sequencing from broth cultures. A total of 197 blaKPC, 171 blaNDM, 142 blaIMP, 6 blaVIM, and 5 blaOXA-48 genes were detected by Xpert Carba-R in 417 rectal swabs, with overall positive and negative percentage agreements ranging from 94.5% to 100% and from 94.8% to 99.9%, respectively. Notably, 17.5% (263/1500) of inpatients had rectal colonization with carbapenem-nonsusceptible organisms detected in intensive care units, and 63.1% (166/263) were Xpert Carba-R positive. Among the 469 carbapenem-nonsusceptible and 52 carbapenem-susceptible isolates examined, 373 were Enterobacteriaceae, 55 were Pseudomonas aeruginosa, and 93 were Acinetobacter baumannii. Compared with the reference isolate sequencing, overall positive and negative percentage agreements were 99.7% and 98.0%, respectively. The intra-assay and interassay coefficient of variability values were both <2%. Thus, we show that Xpert Carba-R assay provides good reproducibility and reliable results for detection and differentiation of five carbapenemase genes in both rectal swabs and clinical isolates.


The first case of Tyzzer's disease in a young foal in Italy: a case report.
Seizures, coma and death rapidly appeared after admission in a one ‑month‑old foal with a history of lethargy, fever and anorexia. Severe icterus and necrotizing hepatitis were observed at necropsy. Clinical signs, laboratory and postmortem findings were compatible with a suspect of clostridial hepatitis. Tyzzer's disease was confirmed by the presence of organisms morphologically consistent with Clostridium piliforme in the hepatocytes at the margins of multiple areas of hepatic necrosis. To the authors' knowledge, this is the first reported case of clostridial hepatitis caused by Clostridium piliforme in a horse in Italy.


Bradykinin contracts the pupillary sphincter and evokes ocular inflammation through release of neuronal substance P.
Bradykinin contracts the isolated rabbit sphincter pupillae muscle. The contraction produced by 10(-8) M bradykinin was resistant to atropine but not to tetrodotoxin, suggesting a non-cholinergic nervous mechanism. The contraction was blocked by specific substance P (SP) antagonists, suggesting the involvement of SP. The SP antagonists tested were [D-Pro2,D-Trp7,9]SP-(1-11) and [Arg5,D-Trp7,9]SP-(5-11). The bradykinin-induced contraction exhibited marked tachyphylaxis in contrast to that induced by SP. It appears that the tachyphylaxis reflects the depletion of a bradykinin-sensitive neuronal pool of SP. Injection of bradykinin into the vitreous chamber of the rabbit eye caused miosis and disruption of the blood-aqueous barrier (manifested as aqueous flare). A second administration of bradykinin a few hours after the first injection evoked a reduced response; the response to SP upon repeated administration was unchanged. Atropine was without effect on the response to bradykinin whereas tetrodotoxin and the SP antagonists reduced the response. The results suggest that bradykinin causes miosis and aqueous flare at least partly through local release of neuronal SP.


An interrupted time series analysis of trends in opioid-related emergency department visits from pre-COVID-19 pandemic to pandemic, from the Canadian Hospitals Injury Reporting and Prevention Program.
Opioid-related emergency department (ED) visits in Canada increased during the COVID-19 pandemic, but how trends in volume and case severity changed from pre-pandemic times through the pandemic is not known. Trends in ED visits related to specific types of opioids also remain unclear. Our objective was to describe pre-pandemic trends and how they changed with the onset of COVID-19 and thereafter. Based on data from the Canadian Hospitals Injury Reporting and Prevention Program, we identified opioid-related ED visits and constructed a time series from March 12, 2018 through March 7, 2021-two pre-COVID periods and one COVID period. We used an interrupted time series (ITS) analysis to examine trends in volume and case severity. We compared medians and means of monthly counts and percentages of severe cases between the periods, by sex, age, and opioid type. Before the pandemic, there was an increasing trend in fentanyl-related visits for males, females and 25- to 64-year-olds, and a decreasing trend in heroin-related visits for males and 18- to 64-year-olds. Fentanyl-related visits for 18- to 24-year-olds showed an immediate increase at the start of the pandemic and a decreasing trend during the pandemic. Heroin-related visits for 12- to 17-year-olds had an immediate increase at the start of the pandemic; for 18- to 24-year-olds and 45- to 64-year-olds, the prior decreasing pre-pandemic trend ceased. For pooled opioid-related visits, no significant trend in the percentage of severe cases was observed throughout the entire study period. This study shows that an ITS approach in trend analysis is a valuable supplement to comparisons of before and after measures (with or without controlling seasonal effects). The findings provide evidence on how ED presentations for opioid use evolved in Canada from 2018 to 2021. The results can inform policies designed to reduce opioid-related harm in the context of a public health emergency.


Serum-based surface-enhanced Raman spectroscopy combined with PCA-RCKNCN for rapid and accurate identification of lung cancer.
Early and precise diagnosis of lung cancer is critical for a better prognosis. However, it is still a challenge to develop an effective strategy for early precisely diagnose and effective treatments. Here, we designed a label-free and highly accurate classification serum analytical platform for identifying mice with lung cancer. Specifically, the microarray chip integrated with Au nanostars (AuNSs) array was employed to measure the surface-enhanced Raman scattering (SERS) spectra of serum of tumor-bearing mice at different stages, and then a recognition model of SERS spectra was constructed using the principal component analysis (PCA)-representation coefficient-based k-nearest centroid neighbor (RCKNCN) algorithm. The microarray chip can realize rapid, sensitive, and high-throughput detection of SERS spectra of serum. RCKNCN based on the PCA-generated features successfully differentiated the SERS spectra of serum of tumor-bearing mice at different stages with a classification accuracy of 100%. The most prominent spectral features for distinguishing different stages were captured in PCs loading plots. This work not only provides a practical SERS chip for the application of SERS technology in cancer screening, but also provides a new idea for analyzing the feature of serum at the spectral level.


Ileocecal bladder augmentation in myelodysplasia.
We discuss 14 children and adolescents with myelodysplasia who underwent bladder augmentation with the ileocecal segment. The bowel was not detubularized nor was the ileocecal valve intussuscepted. Urodynamic evaluation was performed before and after the procedure in 13 patients with a followup of 1 to 8 years. Postoperative capacity and compliance were normal but cecal contractions occurred in 8 patients despite adequate doses of anticholinergics. Reflux was demonstrated at capacity with a cecal contraction in 4 patients but upper tract dilatation and infection were not clinical problems. Three patients required reoperation for complications owing to ureteroileal stenosis and/or urinary tract calculi. Although the clinical results were satisfactory, detubularized segments of bowel with intussuscepted afferent loop valves to prevent reflux may resolve these problems in the future.


Prothrombin and intraventricular haemorrhage.
In the study by Salonvaara et al., low prothrombin activity in preterm infants was associated with intraventricular haemorrhage (IVH). The interesting research question-do preterm infants who develop IVH have underlying coagulation disturbance?-could not be answered in this study. The association needs further investigation. The role of prothrombin in perinatal brain damage in the development of IVH and of white matter damage has to be further elucidated.


Single-wavelength sO2 measurement based on Grüneisen-relaxation photoacoustic effect.
The photoacoustic effect-based sO2 measurement is attracting more and more attention due to its non-invasiveness and accuracy. Compared with the linear dual-wavelength method, the sO2 measurement based on single-wavelength excitation can be potentially applied with simplified system construction. However, the single-wavelength methods proposed in previous studies decreases the safety or lacks the in-depth resolution. This paper proposes a novel single-wavelength method based on the Grüneisen-relaxation (GR) nonlinear effects. It avoids the high fluence excitation with maintaining in-depth resolution and obtains the signals in hundreds of nanoseconds, simultaneously improving the safety and detection speed. The construction of a single laser source for GR effect generation makes the system stable. The sO2 quantification results of blood samples have a good consistency with the reference values. Our work provides a safer and faster measurement method, and a stable system, to promote its application in the clinical area.


Haemodynamic and blood gas changes in buffaloes (Bubalus bubalis) in the supine position following thiopentone anaesthesia with premedication.
Studies were undertaken on seven adult female buffaloes (Bubalus bubalis) premedicated with triflupromazine hydrochloride (0.3 mg/kg) and chloral hydrate (30 mg/kg). Anaesthesia was induced with thiopentone sodium (10 mg/kg) and animals were kept under intermittent positive pressure ventilation. Measurements were made for 30 min in the lateral position, for 20 min in the supine position and subsequently for 40 min again in the lateral position. Regurgitation during anaesthesia was avoided by evacuation of the rumen by rumenotomy two days prior to the experiment. When animals were in the lateral position there were minimal changes in arterial pressure and heart rate but central venous pressure rose. These changes were related to intermittent positive pressure ventilation. In the supine position, increase in heart rate (28%) was associated with profound hypotension (56%) and decrease in central venous pressure (171%) in these animals. There was a fall in venous oxygen tension and a marked increase (91%) in oxygen extraction ratio. Restoration to the lateral position ameliorated all these changes rapidly. It was suggested that a 'low pressure-low flow' circulatory state exists in anaesthetised adult buffaloes in the supine position.


Microfluidic assay without blocking for rapid HIV screening and confirmation.
The essential step for HIV spreading limitation is the screening tests. However, there are multiple disadvantages in current screening assays which need further confirmation test. Herein we developed a rapid HIV assay combining screening and confirmation test by using the microfluidic network assay. Meanwhile, the assay is accelerated by bypassing the step of blocking. We call this method as microfluidic assay without blocking (MAWB). Both the limit of detection and reagent incubation time of MAWB are determined by screening of one model protein pair: ovalbumin and its antibody. The assay time is accelerated about 25% while the limit of detection (LOD) is well kept. Formatting the method in for both HIV screening (testing 8 HIV-related samples) and confirmation (assaying 6 kinds of HIV antibodies of each sample) within 30 min was successful. Fast HIV screening and confirmation of 20 plasma samples were also demonstrated by this method. MAWB improved the assay speed while keeping the LOD of conventional ELISA. Meanwhile, both the accuracy and throughput of MAWB were well improved, which made it an excellent candidate for a quick HIV test for both screening and confirmation. Methods like this one will find wide applications in clinical diagnosis and biochemical analysis based on the interactions between pairs of molecules.


Early tracheostomy in patients undergoing mechanical thrombectomy for acute ischemic stroke.
Respiratory failure following mechanical thrombectomy (MT) for acute ischemic stroke (AIS) is a known complication, and requirement of tracheostomy is associated with worse outcomes. Our objective is to evaluate characteristics associated with tracheostomy timing in AIS patients treated with MT. The National Inpatient Sample was queried for adult patients treated with MT for AIS from 2016 to 2019. Baseline demographic characteristics, comorbidities, and inpatient outcomes were analyzed for associations in patients who received tracheostomy. Timing of early tracheostomy (ETR) was defined as placement before day 8 of hospital stay. Of 3505 AIS-MT patients who received tracheostomy, 915 (26.1%) underwent ETR. Patients who underwent ETR had shorter length of stay (LOS) (25.39 days vs 32.43 days, p < 0.001) and lower total hospital charges ($483,472.07 vs $612,362.86, p < 0.001). ETR did not confer a mortality benefit but was associated with less acute kidney injury (OR, 0.697; p = 0.013), pneumonia (OR, 0.449; p < 0.001), and sepsis (OR, 0.536; p = 0.002). An expected increase in complications and healthcare resource utilization is seen in AIS-MT patients receiving tracheostomy, likely reflecting the severity of patients' post-stroke neurologic injury. Among these high-risk patients, ETR was predictive of shorter LOS and fewer complications.


Sporulation of Bacillus subtilis in Binary Cultures with Ultramicrobacteria.
The effect of ultramicrobacterial epibionts of the genera Kaistia (strain NF1), Chryseobacterium (strain NF4), and Stenotrophomonas (strain FM3) on the process of sporulation of Bacillus subtilis ATCC 6633 was studied. The investigated strains of ultramicrobacteria (UMB) were found to inhibit the sporulation process of B. subtilis ATCC 6633 in binary mixed cultures, exhibiting a 3-day delay of the onset of sporulation compared to the control one, an extended period of the prospore maturation, formation of the fraction of immature spores, and development of ultrastructural defects in many endospores. Thus, investigation of binary mixed cultures of B. subtilis and UMB revealed that, apart from suppression of reproduction and lysis of host vegetative cells, inhibition of spore formation and destruction of endospores was yet another feature of intermicrobial parasitism. The UMB parasites of the studied genera are assumed to participate in the regulation of development and reproduction of B. subtilis in natural habitats of this spore-forming bacterium.


Modified forms of vasopressin and oxytocin in a bovine pineal preparation.
A bovine pineal acid extract displays a vasotocin-like bioactivity in several bioassays, and is recognized by antibodies against the Pro-Arg-Gly-amide ending common to vasopressin and vasotocin. By using molecular sieve filtration and reversed-phase HPLC, a vasopressin- and oxytocin-like peptide was isolated from this pineal preparation, while no evidence for a vasotocin-like peptide was obtained. The isolated neuropeptides contain a modified amino acid at position 2. This structural difference with authentic pituitary vasopressin and oxytocin may alter their biological and immunological properties, which have been interpreted as vasotocin-like, and thus underlies the controversy concerning the existence of vasotocin in the mammalian pineal gland.


Mitochondrial DNA induces nucleus pulposus cell pyroptosis via the TLR9-NF-κB-NLRP3 axis.
Nucleus pulposus cell (NPC) death and progressive reduction play important roles in intervertebral disc degeneration (IVDD). As part of a damage-associated molecular pattern, mitochondrial DNA (mtDNA) can be recognized by TLR9 and triggers the expression of NF-κB and NLRP3 inflammasomes, inducing pyroptosis and inflammatory response. However, whether mtDNA induces NPC pyroptosis via the TLR9-NF-κB-NLRP3 axis and promotes IVDD remains uncertain. We constructed an in vitro NPC oxidative stress injury model to clarify the mechanism of mtDNA release, TLR9-NF-κB signaling pathway activation, and NPC injury. We further verified the mechanism of action underlying the inhibition of mtDNA release or TLR9 activation in NPC injury in vitro. We then constructed a rat punctured IVDD model to understand the mechanism inhibiting mtDNA release and TLR9 activation in IVDD. We used human NP specimen assays to show that the expression levels of TLR9, NF-κB, and NLRP3 inflammasomes correlated with the degree of IVDD. We demonstrated that mtDNA mediated TLR9-NF-κB-NLRP3 axis activation in oxidative stress-induced human NPC pyroptosis in vitro. Oxidative stress can damage the mitochondria of NPCs, causing the opening of the mitochondrial permeability transition pores (mPTP) and leading to the release of mtDNA into the cytosol. Furthermore, inhibition of mPTP opening or TLR9 activation blocked TLR9-NF-κB-NLRP3 axis activation and thereby mediated NPC pyroptosis and IVDD. mtDNA plays a key role in mediating NPC pyroptosis and IVDD via the TLR9-NF-κB-NLRP3 axis. Our findings provide new potential targets for IVDD.


A Case of Urbanorum spp. in a Woman from an Urban-Marginal Sector of Ecuador.
BACKGROUND Worldwide, there are few cases of Urbanorum spp. in humans; however, it is associated with gastrointestinal pathologies, where humans probably acquire the disease by fecal-oral transmission, by ingesting food or water with infective cysts. The main symptoms of the patients who have this presumed parasite are fever, vomiting, colic, dyspepsia, and watery diarrhea. Since the first case of Urbanorum spp. was reported in 1994 in Colombia, cases have subsequently been reported in Peru, Ecuador, Colombia, Brazil, and Mexico. In Ecuador, a prevalence of 1.16% has been reported, and the objective of this study was to record another case of Urbanorum spp. infection in Ecuador. CASE REPORT A female patient (mixed race; 40 years old; and thin, weighing 57 to 62 kg) requested help from the FCI Project approved and financed by the University of Guayaquil (Ecuador). She underwent routine tests, such as direct parasitology and sedimentation with centrifugation using saline solution. Samples were observed under an optical microscope with 10x and 40x objectives and stained with and without Lugol's solution. We detected a rounded structure with several filaments similar to light yellow pseudopods. CONCLUSIONS Herein, a case of Urbanorum spp. infection in Ecuador, where current environmental and sanitary conditions have contributed to new cases, is reported, indicating that the community was exposed to this probable parasite with importance in public health. Further studies are recommended to confirm its etiology, life cycle, and epidemiology, in order to create a national registry, in case it is defined as a protozoan.


Effect of dapagliflozin on COVID-19 infection and risk of hospitalization.
Dapagliflozin has been proposed as a potential treatment for coronavirus disease 2019 (COVID-19) by reducing cytokine production and inflammation. However, there are limited data on its effectiveness. We aimed to evaluate the impact of dapagliflozin on COVID-19 severity (including hospitalization risk, ICU admission, in-hospital death and progression to severe COVID-19) and its potential on susceptibility to COVID-19 infection. We conducted a population-based case-control study. For aim 1, we assessed COVID-19 severity in cases (positive PCR patients requiring hospitalization) and matched controls (negative PCR patients or positive PCR patients not requiring hospitalization). For aim 2, we compared positive PCR cases (hospitalized and non-hospitalized) with controls. Adjusted odds ratios (aORs) were calculated using a generalized linear mixed model. We analysed 86 602 subjects: 3060 were hospitalized cases, 26 757 were non-hospitalized cases and 56 785 were controls. Among the hospitalized COVID-19 patients, 228 were admitted to the ICU and 413 died. Dapagliflozin had no effect on the risk of hospitalization (aOR 0.98; 95% CI 0.65-1.48; P = 0.915), ICU admissions (aOR 1.21; 95% CI 0.34-4.25; P = 0.767) or in-hospital death (aOR 1.33; 95% CI 0.53-3.30; P = 0.543). Dapagliflozin reduced the risk of progression to severe COVID-19 by 35%, but this was not statistically significant (aOR 0.65; 95% CI 0.40-1.06; P = 0.086). Dapagliflozin was associated with a 30% increased risk of susceptibility to COVID-19 infection (aOR 1.31; 95% CI 1.05-1.62; P = 0.015). Use of dapagliflozin prior to SARS-CoV-2 infection was not associated with an increased risk of hospitalization, ICU admission, mortality or progression to severe COVID-19. However, it was associated with an increased risk of susceptibility to COVID-19 infection.


Immunological consequences of intestinal helminth infections: antigen presentation and immunosuppression by peritoneal cells.
The functional characteristics of peritoneal cells (PC's) acquired in response to infection with Nematospiroides dubius were studied to identify changes which could be associated with the high level, long-term survival of this parasite or explain its transient immunosuppressive properties. The number of cells in the peritoneal cavity increased steadily during the infection, and the population displayed a transient hyperresponsiveness to inflammatory stimulation. N. dubius did not affect the ability of antigen-pulsed PC's to induce humoral responses or delayed-type hypersensitivity (DTH) in primed recipients, but lymphoproliferative responses induced in vitro were depressed by PC's from infected donors. In addition, unpulsed PC's from N. dubius-infected mice depressed proliferative responses to parasite antigens by autologous mesenteric lymph node cells. These effects are attributable to increased lymphocytostatic activity in the PC's, which peaked one week after infection and correlated with the expansion of a population of large plastic-adherent vacuolated cells with elevated acid phosphatase activity (activated macrophages). Comparable inhibitory cells did not develop in the same period of time during a short-lived infection with Nippostrongylus brasiliensis.


Effective, safe and affordable - the nurse's dream shopping list.
A team of nurses is being charged with the task of assessing equipment for cost and clinical effectiveness in a bid to improve NHS procurement.