First Molecular Genotyping of Cryptosporidium felis in Cattle, Iraq.

To the best of our knowledge, this is the first Iraqi study to detect C. felis in cattle by the polymerase chain reaction (PCR) assay and to confirm the local isolates in the National Center for Biotechnology Information (NCBI). Overall, 130 diarrheic calves of different ages and sexes were selected randomly from rural and sub-urban areas in Wasit province (Iraq) from February to April (2021) and subjected to direct collection of fresh fecal samples for DNA extraction and PCR examination. Targeting the heat shock protein 70 (HSP70) gene of C. felis showed that 17.69% of the animals were positive. Findings from clinical examination revealed no significant differences in values of temperature, pulse, or respiratory rates between the positive and negative calves. The association between the positive results and demographic risk factors showed no significant differences in the prevalence rate of infection and risk of exposure to C. felis between the rural and sub-urban areas; however, higher significant values were reported in calves aged 6 months than in calves aged 12 months, as well as in females than in males. Five of the local C. felis isolates were documented under the accession numbers MZ964600.1, MZ964601.1, MZ964602.1, MZ964603.1, and MZ964604.1. Finally, the data presented here provide epidemiological and molecular evidence that the range of C. felis in cattle is wider than expected globally, with a high probability of infection transmission from cattle to humans.

Revised recommendations from the CSO-HNS taskforce on performance of tracheotomy during the COVID-19 pandemic - what a difference a year makes.

BACKGROUND: During the early part of the COVID-19 pandemic, the Canadian Society of Otolaryngology - Head & Neck Surgery (CSO-HNS) task force published recommendations on performance of tracheotomy. Since then, our understanding of the virus has evolved with ongoing intensive research efforts. New literature has helped us better understand various aspects including patient outcomes and health care worker (HCW) risks associated with tracheotomy during the COVID-19 pandemic. Accordingly, the task force has re-evaluated and revised some of the previous recommendations. MAIN BODY: Based on recent evidence, a negative reverse transcription polymerase chain reaction (RT-PCR) COVID-19 swab status is no longer the main deciding factor in the timing of tracheotomy. Instead, tracheotomy may be considered as soon as COVID-19 swab positive patients are greater than 20 days beyond initial symptoms and 2 weeks of mechanical ventilation. Furthermore, both open and percutaneous surgical techniques may be considered with both techniques showing similar safety and outcome profiles. Additional recommendations with discussion of current evidence are presented. CONCLUSION: These revised recommendations apply new evidence in optimizing patient and health care system outcomes as well as minimizing risks of COVID-19 transmission during aerosol-generating tracheotomy procedures. As previously noted, additional evidence may lead to further evolution of these and other similar recommendations.

Recommendations from the CSO-HNS taskforce on performance of tracheotomy during the COVID-19 pandemic.

INTRODUCTION: The performance of tracheotomy is a common procedural request by critical care departments to the surgical services of general surgery, thoracic surgery and otolaryngology - head & neck surgery. A Canadian Society of Otolaryngology - Head & Neck Surgery (CSO-HNS) task force was convened with multi-specialty involvement from otolaryngology-head & neck surgery, general surgery, critical care and anesthesiology to develop a set of recommendations for the performance of tracheotomies during the COVID-19 pandemic. MAIN BODY: The tracheotomy procedure is highly aerosol generating and directly exposes the entire surgical team to the viral aerosol plume and secretions, thereby increasing the risk of transmission to healthcare providers. As such, we believe extended endotracheal intubation should be the standard of care for the entire duration of ventilation in the vast majority of patients. Pre-operative COVID-19 testing is highly recommended for any non-emergent procedure. CONCLUSION: The set of recommendations in this document highlight the importance of avoiding tracheotomy procedures in patients who are COVID-19 positive if at all possible. Recommendations for appropriate PPE and environment are made for COVID-19 positive, negative and unknown patients requiring consideration of tracheotomy. The safety of healthcare professionals who care for ill patients and who keep critical infrastructure operating is paramount.

Determination of the Effective Dose of Curcumin alone and in Combination with Antimicrobial Peptide CM11 on Promastigote Forms of Iranian Strain of L. major (MRHO / IR / 75 / ER).

Zoonotic cutaneous leishmaniasis t caused by Leishmania major is spread in focal areas of more than 90 countries in the tropics, subtropics, and southern Europe. In the absence of any effective vaccine, the only means to treat and control leishmaniasis is conventional medication. Glucantime is the first choice of anti-leishmanialdrug, has serious side effects like high toxicity, exorbitant cost, problems with the administration and development of resistance. Curcumin is the active component from the rhizome of herb Curcuma longa, possessing many pharmacological and biological activities with antiprotozoal and anti-proliferative effects which make it a good alternative to existing therapy. Antimicrobial peptides like CM11, a small peptide consisting of 11 amino acids, are also novel potential drugs against at least wide spectrum of microbial organisms. The aim of this study was to evaluate the effect of curcumin alone and in combination with CM11 on promastigote form of L. major (MRHO / IR / 75 / ER) for 12h and 24h in vitro. The results of Giemsa staining showed that the morphology of the flagellum and cell shape increased changed with increasing concentration of curcumin (5 µM, 10 μM, 20 μM, 40 μM and 80 μM). MTT and Trypan blue results demonstrated that the promastigotes were susceptible against curcumin in dose and time dependent manner, while CM11 alone at concentration of 8 µM as well as in combination with 10 and 20 µM curcumin had no significant effect on promastigotes. Our results revealed that curcumin can provide a new curative candidate against cutaneous leishmaniasis.

Epidemiological evaluation of Leishmania infantum zoonotic transmission risk in the recently established endemic area of Northwestern Italy.

Leishmania infantum infection had been expanding into new areas due to changes in vector and host biology. Zoonotic visceral leishmaniasis has become endemic in previously unsuitable areas as vectors find favourable climatic conditions and an increasing number of reservoir dogs are moved between traditionally and new endemic areas. Monitoring vector and disease expansion in areas of recent colonization is needed to understand transmission mechanisms and patterns of disease establishment. Here, we studied the infection status of 815 human blood donors and of 803 sympatric dogs from five, newly endemic, areas in Northwestern Italy. In autochthonous dogs, the seroprevalence of anti-L. infantum antibodies, recorded by Western blot, reached 42.22%, while in humans, the seroprevalence was of 16.81%. No significant correlation between the infection status of dogs and that of their human owners was found, but L. infantum infection was recorded in the different study areas with significant levels of diversity. Restriction fragment length polymorphism showed a high genetic variability of the circulating strains and gave useful insights on patterns of disease establishment into a naïve area.

[3D structure of DKK1 indicates its involvement in both canonical and non-canonical Wnt pathways].

Dikkoppf-1 (DKK1) is an antagonist of the canonical Wnt signaling pathway. The importance of DKK1 as a diagnostic and therapeutic agent in a wide range of diseases along with its significance in a variety of biological processes accentuate the necessity to decipher its 3D structure that would pave the way towards the development of relevant selective inhibitors. A DKK1 structure model predicted by the Robetta server with structural refinements including a 10 ns molecular dynamics run was subjected to functional and docking analyses. We hypothesize that the N-terminal region of the DKK1 molecule could be functionally important for both canonical and noncanonical Wnt pathways. Moreover, it seems that DKK1 could be involved in interactions with the Frizzled receptors, leading to the activation of the Planar Cell Polarity (PCP) pathway (activation of Jun N-terminal kinase (JNK) Pathway) and Wnt/Ca^(2+) pathway (activation of CamKII).

Perfluorodecalin and bone regeneration.

Perfluorodecalin (PFD) is a chemically and biologically inert biomaterial and, as many perfluorocarbons, is also hydrophobic, radiopaque and has a high solute capacity for gases such as oxygen. In this article we have demonstrated, both in vitro and in vivo, that PFD may significantly enhance bone regeneration. Firstly, the potential benefit of PFD was demonstrated by prolonging the survival of bone marrow cells cultured in anaerobic conditions. These findings translated in vivo, where PFD incorporated into bone-marrow-loaded 3D-printed scaffolds substantially improved their capacity to regenerate bone. Secondly, in addition to biological applications, we have also shown that PFD improves the radiopacity of bone regeneration biomaterials, a key feature required for the visualisation of biomaterials during and after surgical implantation. Finally, we have shown how the extreme hydrophobicity of PFD enables the fabrication of highly cohesive self-setting injectable biomaterials for bone regeneration. In conclusion, perfluorocarbons would appear to be highly beneficial additives to a number of regenerative biomaterials, especially those for bone regeneration.

High levels of HCV core+1 antibodies in HCV patients with hepatocellular carcinoma.

The core region of the hepatitis C virus (HCV) genome possesses an overlapping ORF that has been shown to encode a protein, known as the alternate reading frame protein (ARFP), F or core+1. The biological role of this protein remains elusive, as it appears to be non-essential for virus replication. However, a number of independent studies have shown that the ARFP/F/core+1 protein elicits humoral and cellular immune responses in HCV-infected individuals and interacts with important cellular proteins. To assess the significance of the core+1 humoral response in HCV-infected patients, we examined the prevalence of anti-core+1 antibodies in sera from patients with hepatocellular carcinoma (HCC) in comparison with chronically HCV-infected individuals without HCC. We produced two HCV core+1 histidine-tagged recombinant proteins for genotypes 1a (aa 11-160) and 1b (aa 11-144), as well as a non-tagged highly purified recombinant core+1/S protein (aa 85-144) of HCV-1b. Using an in-house ELISA, we tested the prevalence of core+1 antibodies in 45 patients with HCC in comparison with 47 chronically HCV-infected patients without HCC and 77 negative-control sera. More than 50 % of the serum samples from HCC patients reacted with all core+1 antigens, whereas <26 % of the sera from the non-HCC HCV-infected individuals tested positive. No core+1-specific reactivity was detected in any of the control samples. In conclusion, the high occurrence of anti-core+1 antibodies in the serum of HCC patients suggests a role for the ARFP/F/core+1 protein in the pathogenesis of HCC.

A Lack of tolerance to the anxiolytic action of Echium amoenum.

The anxiolytic effect of the flower of Echium. amoenum was shown in several experimental studies in mice. The present study was aimed to determine whether tolerance develops to anxiolytic action of E. amoenum in mice. NMRI male mice were injected intraperitoneal with hydroalcoholic extract (12.5, 25 and 50 mg/kg) or saline once each day (8 am) for period of 7 days and then tested on light/dark box model. Anxiolytic effect was determined by light/dark box and elevated plus-maze. According to the results, hydroalcohoic extract of E. amoenum when given both acutely and chronically (7 days) at 25 and 50 mg/kg, significantly increased the time in the illuminated zone. The number of transitions in the light/dark apparatus, however, was not significantly altered by the tested doses of the plant. Diazepam at 0.5 and 1 mg/kg produced anxiolytic effect in both model of anxiety, namely, the light/dark box and elevated plus-maze. No tolerance was developed to the anxiolytic effect of E. amoenum extract after 7 days of treatment. Our results suggest that one week treatment with extract of the E. amoenum does not produce tolerance to its anxiolytic action. Longer period of treatment using implant procedure is probably necessary to cause molecular changes in order to induce tolerance.

Gender similarity in low agreement between written and video ISAAC asthma questionnaires.

BACKGROUND AND AIM: The prevalence of childhood asthma varies among different nations and appears to have increased in recent years. It has been difficult to determine this prevalence precisely and compare the communities due to the lack of a standardized method. The International Studies of Asthma and Allergies in Childhood (ISAAC) has developed a written and video questionnaire to measure asthma prevalence and overcome these difficulties. This study was carried out to determine the prevalence of childhood asthma in Kashan, the central area of Iran, and to assess the agreement between the ISAAC written and video questionnaires and its gender relationship. METHODS: Between December 2002 and June 2003, 2533 school children, aged 13-14 years, in Kashan secondary schools completed sequentially the ISAAC written and video questionnaires. The agreement between responses to the two questionnaires for reported "ever wheeze", "exercise wheeze", "night wheeze" and "night cough" was determined, using kappa coefficient and concordance. RESULTS: The prevalence of asthma symptoms were significantly lower in the responses to the video questionnaire compared with the written questionnaire. Kappa coefficient showed only poor to fair agreement (kappa = 0.13-0.24) between video and written questionnaires, although the concordance between them always exceeded 60%. CONCLUSIONS: This study showed that reported asthma symptoms based on video questionnaires were significantly lower than the written questionnaire, and that there is low agreement between the two questionnaires which requires further investigation to explain the findings. In addition, this investigation revealed no significant difference between the genders for this low agreement.

Identification, cloning, and initial characterization of rot, a locus encoding a regulator of virulence factor expression in Staphylococcus aureus.

A chromosomal insertion of transposon Tn917 partially restores the expression of protease and alpha-toxin activities to PM466, a genetically defined agr-null derivative of the wild-type Staphylococcus aureus strain RN6390. In co-transduction experiments, transposon-encoded erythromycin resistance and a protease- and alpha-toxin-positive phenotype are transferred at high frequency from mutant strains to agr-null strains of S. aureus. Southern analysis of chromosomal DNA and sequence analysis of DNA flanking the Tn917 insertion site in mutant strains revealed that the transposon interrupted a 498-bp open reading frame (ORF). Similarity searches using a conceptual translation of the ORF identified a region of homology to the known staphylococcal global regulators AgrA and SarA. To verify that the mutant allele conferred the observed phenotype, a wild-type allele of the mutant gene was introduced into the genome of a mutant strain by homologous recombination. The resulting isolates had a restored agr-null phenotype. Virulence factor gene expression in mutant, restored mutant, and wild-type strains was quantified by measuring alpha-toxin activity in culture supernatant fluids and by Northern analysis of the alpha-toxin transcript. We named this ORF rot (for repressor of toxins) (GenBank accession no. AF189239) because of the activity associated with rot::Tn917 mutant strains.

High incidence of penicillin resistance amongst clinical isolates of Streptococcus pneumoniae in northern Palestine.

One hundred and thirteen consecutive isolates of Streptococcus pneumoniae were collected in Nablus, Palestine between March and Aug. 1997 from children with acute lower respiratory tract infections. Resistance rates were: penicillin 88%, cefuroxime 85%, erythromycin 63%, tetracycline 45%, chloramphenicol 27% and ofloxacin 2%. Resistances to erythromycin and cefuroxime were significantly associated with penicillin resistance. Ofloxacin may be useful against pneumococci resistant to traditional antimicrobial agents. Factors associated with penicillin resistance included hospitalisation and previous use of beta-lactam antibiotics.

FT-IR microspectroscopic imaging of flax (Linum usitatissimum L.) stems.

Fourier transform infrared (FT-IR) microspectroscopy was used to image and subsequently produce maps of the distribution chemical components in flax stem tissue. Cross-sections of two flax cultivars, Ariane and Natasja, were examined. The infrared maps produced were keyed to the visible images of the samples, thus permitting superimposition of chemical composition onto anatomical structure. The functional group frequencies utilized to produce chemical maps were based on their diagnostic capability for indicating each component to the exclusion of others in the matrix, by comparison of the spectra of "pure" components. Waxes were best indicated by a sharp shoulder at 2850 cm(-1) and appeared primarily in the cuticular and epidermal tissues. Pectins gave a band ca. 1615 cm(-1) for the Ca2+ salt form that was primarily detected in tissue surrounding the fiber bundles. Cellulose was the strongest contributor to the band located at ca. 1335 cm(-1) that showed its greatest intensity in the fiber cells. Aromatic compounds were indicated most readily by the band at 1510 cm(-1) and were located primarily in the core tissue. Acetyl groups, likely associated with hemicellulosic polysaccharides, were indicated by a band centered at 1250 cm(-1) and found within the fibers and core tissue.

Aedes aegypti: induced antibacterial proteins reduce the establishment and development of Brugia malayi.

The effect of host immune activation on the development of Brugia malayi in one susceptible and four refractory strains of Aedes aegypti and in Armigeres subalbatus was assessed. A. aegypti that were immune activated by the injection of saline or bacteria 24 hr before feeding on a B. malayi-infected gerbil had significantly reduced prevalences and mean intensities of infection from those of naive controls when exposed to bloodmeals with low (105 mf/20 microliters) and medium (160 mf/20 microliters) microfilaremias. At a higher microfilaremia (237 mf/20 microliters) there were no significant differences in mean intensities, suggesting that the number of parasites ingested may affect the host's ability to mount an effective defense response. Because the major immune proteins in A. aegypti are defensins, we did Northern analyses of fat body RNA 8 hr after immune activation or bloodfeeding. All mosquitoes demonstrated rapid transcriptional activity for defensins following immune activation by intrathoracic inoculation with either saline or bacteria. However, no strain of A. aegypti, susceptible or refractory to B. malayi, nor Ar. subalbatus produced defensin transcripts after bloodfeeding on an uninfected or a B. malayi-infected gerbil. These data suggest that inducible immune proteins of mosquitoes can reduce the prevalence and mean intensity of infections with ingested parasites, but these proteins are not expressed routinely after parasite ingestion and midgut penetration and probably do not contribute to existing refractory mechanisms. Immune proteins such as defensins, however, represent potential candidates to genetically engineer mosquitoes for resistance to filarial worms.