Prevalence of resistance genes and antibiotic resistance profile among Stenotrophomonas maltophilia isolates from hospitalized patients in Iran.

Stenotrophomonas maltophilia has emerged as an important nosocomial pathogen. Treatment of S. maltophilia infections is difficult due to increasing resistance to multiple antibacterial agents. In this 12-month cross-sectional study, from 2017 to 2018, 117 isolates were obtained from different clinical sources and identified by conventional biochemical methods. Antibiotic susceptibility tests were performed according to CLSI 2018. Minocycline disk (30 µg) and E-test strips for ceftazidime, trimethoprim-sulfamethoxazole and chloramphenicol were used. PCR confirmed isolates. The frequency of different classes of integrons (I, II) and resistance gene cassettes (sul1, sul2, dfrA1, dfrA5 and aadB) were determined by PCR. The results showed the highest frequency of resistance to chloramphenicol and ceftazidime with 32 cases (27.11%). Among strains, 12 cases (10.25%) were resistant to trimethoprim-sulfamethoxazole (the lowest frequency of resistance), while 19 (16.1%) isolates were resistant to minocycline. Frequency of sul1, int1, aadB, sul2, dfrA5 genes were 64 (55.08%), 26 (22.3 %), 18 (15.25%) and 17 (14.4%), 14 (11.86%), respectively. int2 and dfrA1 were not detected. Although we have not yet reached a high level of resistance to effective antibiotics such as trimethoprim-sulfamethoxazole, as these resistances can be carried by a plasmid, greater precision should be given to the administration of these antibiotics.

COVID-19 meningitis without pulmonary involvement with positive cerebrospinal fluid PCR.

BACKGROUND: Coronavirus disease 2019 (COVID-19) typically presents with respiratory illness ranging in severity. Neurological complications of the disease remain largely unknown. Herein, we discuss the case of a woman diagnosed with COVID-19 meningitis following two positive cerebrospinal fluid (CSF) RT-PCR assays, and highlight the importance of recognizing the neurological manifestations of the disease. CASE REPORT: The patient was a 49-year-old woman with a history of hypertension who presented with non-specific symptoms (fever, headache, malaise, nausea/vomiting). Chest computed tomography (CT) revealed a lack of pulmonary involvement and oropharyngeal/nasopharyngeal RT-PCR was negative for COVID-19. A lumbar puncture was performed on the third day of admission and the CSF analysis elucidated a viral pattern, but the CSF bacterial culture and RT-PCR assay for herpes simplex virus were both negative. Surprisingly, the CSF RT-PCR for COVID-19 was positive. The diagnosis of COVID-19 meningitis was made and the patient was treated solely with Kaletra® , with a second CSF analysis confirming our unique finding 1 week later. The patient's clinical characteristics improved progressively, and she was discharged in excellent general condition after 21 days. CONCLUSION: In contrast to what was originally believed, the SARS-CoV-2 can cause meningitis in isolation, perhaps by crossing the blood-brain barrier. Hence, it seems essential that physicians maintain a high index of suspicion for neurological involvement among COVID-19 patients, with early CSF analysis and brain imaging sometimes being indicated.

Universal features of the jamming phase diagram of wet granular materials.

We investigate the influence of the shape of a particle on the structure of the jamming phase diagram of wet granular materials. We compute the jamming phase diagram of wet dimers (two fused disks) and compare it with that of the wet disks. Amplitude of the external force at solidification, i.e., the jamming force F(s), is computed as a function of the packing fraction ϕ, the capillary bridge energy ɛ, and the aspect ratio of dimers α. Based on data collapse, an equation for amplitude of the external force at solidification F(s)(ϕ,ɛ,α) is derived. F(s) has scaling and logarithmic relations with ϕ and ɛ, respectively, exactly the same type reported for wet disks earlier. Interestingly, F(s) does not depend on the aspect ratio of dimers α. The only difference is that wet dimers are found to be more stiffer than wet disks. However, the similarities of the equations describing F(s)(ϕ,ɛ,α) of wet dimers and disks imply that there exists, yet unknown, universal aspects of mechanical response of wet granular materials to the external forces, independent from the particle shape. In addition, we study local orientation of particles and its statistical properties.

Growth, weaning performance and blood indicators of humoral immunity in Holstein calves fed supplemental flavonoids.

The primary objective was to test the hypothesis that flavonoids mediate immune response and affect calf performance. Twenty Holstein calves [7 +/- 2 days age; 41.4 +/- 0.7 kg body weight (BW)] were randomly assigned to four treatments of (i) no; (ii) low (7.3 x 10(-5) g/kg BW); (iii) medium (7.3 x 10(-4) g/kg BW); and (iv) high (3.6 x 10(-3) g/kg BW) doses of flavonoids intake in a completely randomized design. Calves received the treatments as a tablet until weaning or a daily intake of 680 g starter. After weaning, calves received no supplemental flavonoids and monitored until 120 days of age. The flavonoids were extracted from propolis. Treatments did not affect body length, wither height and the severity of scours. At week 5 of age, BW was higher when calves fed the high compared to the low dose of flavonoids. At week 6, calves fed the high dose of flavonoids had higher BW than those fed no or low doses of flavonoids. The serum immunoglobulin G (IgG) concentrations remained lower at the first 3 weeks of the experiment when calves received the low but not the high doses of flavonoids. At week 4, both medium and low doses of flavonoids moderated serum IgG. At week 8, the medium and high but not the low doses of flavonoids lowered serum IgG. At week 6, calves fed high and medium flavonoids doses had lower blood immunoglobulin M (IgM) than control calves. Results suggest that flavonoids affect the humoral immune response and can improve growth in young calves. This response depended on calf age. Future studies are needed to further evaluate the premise that dietary forages or the main source of flavonoids are helpful for a less stressful weaning in the modern calf raising.

Quantitative imaging of gene induction in living animals.

Methods to repeatedly, non-invasively, and quantitatively image gene expression in living animals are rapidly emerging and should fundamentally change studies of gene expression in vivo. We previously developed assays utilizing positron emission tomography (PET) to image reporter gene expression. In this paper we: (1) describe a new bi-directional, tetracycline-inducible system that can be used to pharmacologically induce target gene expression and to quantitatively image induced expression by using a PET reporter gene; (2) demonstrate the potential of this system in transient and stable cell transfection assays; and (3) demonstrate the ability to repetitively and quantitatively image tetracycline and tetracycline analog induction of gene expression in living animals. We utilize the dopamine type-2 receptor (D(2)R) and the mutant herpes-simplex virus type 1 thymidine kinase (HSV1-sr39tk) reporter genes to validate this system. We utilize microPET technology to show that quantitative tomographic imaging of gene induction is possible. We find a high correlation (r(2) = 0.98) between 'target' and reporter gene expression. This work establishes a new technique for imaging time-dependent variation of gene expression both from vectors with inducible promoters and in transgenic animals in which pharmacologic induction of gene expression must be monitored. These techniques may be applied both in gene therapy and for the study of gene expression in transgenic animals.

Human pharmacokinetic and dosimetry studies of [(18)F]FHBG: a reporter probe for imaging herpes simplex virus type-1 thymidine kinase reporter gene expression.

UNLABELLED: 9-[4-[(18)F]fluoro-3-(hydroxymethyl)butyl]guanine ([(18)F]FHBG) has been used as a reporter probe to image expression of herpes simplex virus type-1 thymidine kinase (HSV1-tk) reporter gene in living animals. Our aim was to study the kinetics, biodistribution, stability, dosimetry, and safety of [(18)F]FHBG in healthy human volunteers, preparatory to imaging patients undergoing HSV1-tk gene therapy. METHODS: [(18)F]FHBG was synthesized with a specific activity of 37,000--444,000 GBq/mmol and a radiochemical purity > 99%. Ten healthy volunteers consented to participate in the study. A transmission scan was obtained before bolus injection of 70.3--229.4 MBq [(18)F]FHBG into a hand vein, followed by dynamic PET imaging with 4 consecutive emission scans. Warmed hand-vein blood was withdrawn at various times after injection for blood time--activity measurements. Electrocardiography, blood pressure, and blood and urine pharmacologic parameters were measured before and after injection of the [(18)F]FHBG tracer (n = 5). The stability of [(18)F]FHBG in the urine was analyzed. Attenuation-corrected images were reconstructed using the ordered-subsets expectation maximization algorithm. Image region-of-interest time-activity data were used with the MIRD program to estimate absorbed radiation dosages. RESULTS: [(18)F]FHBG had rapid blood clearance; only 8.42% +/- 4.76% (mean +/- SD) of the peak blood activity remained at approximately 30 min. The average ratio of plasma activity to whole-blood activity during the study was 0.91 +/- 0.04. Penetration of [(18)F]FHBG across the blood-brain barrier was not observed. The primary routes of clearance were renal and hepatobiliary. High activities were observed in the bladder, gut, liver, and kidneys, but <0.0002% of the injected dose per gram was observed in other tissues. In the urine, 83% of activity 180 min after injection was stable [(18)F]FHBG. Blood and urine pharmacologic parameters did not change significantly after injection of the [(18)F]FHBG tracer. The bladder absorbed the highest radiation dose. CONCLUSION: [(18)F]FHBG has the desirable in vivo characteristics of stability, rapid blood clearance, low background signal, biosafety, and acceptable radiation dosimetry in humans. This study forms the foundation for using [(18)F]FHBG in applications to monitor HSV1-tk reporter gene expression.

Direct correlation between positron emission tomographic images of two reporter genes delivered by two distinct adenoviral vectors.

Biodistribution, magnitude and duration of a therapeutic transgene's expression may be assessed by linking it to the expression of a positron emission tomography (PET) reporter gene (PRG) and then imaging the PRG's expression by a PET reporter probe (PRP) in living animals. We validate the simple approach of co-administering two distinct but otherwise identical adenoviruses, one expressing a therapeutic transgene and the other expressing the PRG, to track the therapeutic gene's expression. Two PET reporter genes, a mutant herpes simplex virus type 1 thymidine kinase (HSV1-sr39tk) and dopamine-2 receptor (D(2)R), each regulated by the same cytomegalovirus (CMV) promoter, have been inserted into separate adenoviral vectors (Ad). We demonstrate that cells co-infected with equivalent titers of Ad-CMV-HSV1-sr39tk and Ad-CMV-D(2)R express both reporter genes with good correlation (r(2) = 0.93). Similarly, a high correlation (r(2) = 0.97) was observed between the expression of both PRGs in the livers of mice co-infected via tail-vein injection with equivalent titers of these two adenoviruses. Finally, microPET imaging of HSV1-sr39tk and D(2)R expression with 9-(4-[(18)F]fluoro-3-hydroxymethylbutyl) guanine ([(18)F]FHBG) and 3-(2-[(18)F]fluoroethyl)spiperone ([(18)F]FESP), utilizing several adenovirus-mediated delivery routes, illustrates the feasibility of evaluating relative levels of transgene expression in living animals, using this approach.

Dietary titanium and infant growth.

Dietary titanium as TiO2+ improved animal growth during infancy while inhibiting the metabolism of intestinal bacteria. TiO2+ was also found capable of inhibiting human cytomegalovirus in tissue culture. These and other findings indicate TiO2+ improves infant growth by acting as an antibacterial and antiviral agent. The behavior of TiO2+ stands in contrast to that of TiO2, which is inert.

Biological roles of titanium.

Ti4+ in soil is a natural antibiotic mobilized by bacteria-generated H+. When added to the diet of young mice, Ti4+ enhanced their growth. These and observations of others indicate that Ti4+ has a variety of biological roles.

Prevalence of myocardial viability as detected by positron emission tomography in patients with ischemic cardiomyopathy.

BACKGROUND: Detection of myocardial viability is important in patients with ischemic cardiomyopathy. Restoration of blood flow to viable myocardium is associated with improved left ventricular function and improved patient prognosis. However, the prevalence of viable myocardium in patients with ischemic cardiomyopathy is unknown. METHODS AND RESULTS: To determine the prevalence of myocardial viability, clinical [13N]ammonia/18F-deoxyglucose PET studies performed in 283 patients (age, 63+/-10 years) with ischemic heart disease (mean ejection fraction, 26+/-8%) were visually analyzed for the presence and extent of viable and nonviable myocardium. The myocardium was divided into 19 segments. The extent of viable myocardium was considered "functionally" significant if >/=5 segments ( approximately 25% of the left ventricular myocardium) exhibited a blood flow/metabolism mismatch and "prognostically" significant if 1 to 4 left ventricular segments did so. Of all patients, 41% had no evidence of viable myocardium, 55% had viable myocardium, and 4% had normal blood flow and metabolism within an enlarged left ventricle. Functionally significant viability was found in 27% and prognostically significant viability in 28% of the patients. Multivariate analysis revealed the presence of angina to be the only clinical parameter associated with the presence of functionally significant viability. CONCLUSIONS: Revascularization might improve patient prognosis in 55% and result in improved left ventricular function in 27% of all patients with ischemic cardiomyopathy.

Glycine-induced long-term potentiation is associated with structural and functional modifications of alpha-amino-3-hydroxyl-5-methyl-4-isoxazolepropionic acid receptors.

Global long-term potentiation (LTP) was induced in organotypic hippocampal slice cultures by a brief application of 10 mM glycine. Glycine-induced LTP was occluded by previous theta burst stimulation-induced potentiation, indicating that both phenomena share similar cellular processes. Glycine-induced LTP was associated with increased [3H]alpha-amino-3-hydroxyl-5-methyl-4-isoxazolepropionic acid (AMPA) binding in membrane fractions as well as increased amount of a selective spectrin breakdown product generated by calpain-mediated spectrin proteolysis. Antibodies against the C-terminal (C-Ab) and N-terminal (N-Ab) domains of GluR1 subunits were used to evaluate structural changes in AMPA receptor properties resulting from glycine-induced LTP. No quantitative or qualitative changes were observed in Western blots from membrane fractions prepared from glycine-treated slices with C-Ab. In contrast, Western blots stained with N-Ab revealed the formation of a 98-kDa species of GluR1 subunits as well as an increased amount of immunoreactivity after glycine-induced LTP. The amount of spectrin breakdown product was positively correlated with the amount of the 98-kDa species of GluR1 after glycine treatment. Functional modifications of AMPA receptors were evaluated by determining changes in the effect of pressure-applied AMPA on synaptic responses before and after glycine-induced LTP. Glycine treatment produced a significant increase in AMPA receptor function after potentiation that correlated with the degree of potentiation. The results indicate that LTP induction produces calpain activation, truncation of the C-Ab domain of GluR1 subunits of AMPA receptors, and increased AMPA receptor function. They also suggest that insertion of new receptors takes place after LTP induction.

Effects of a nitric oxide synthase inhibitor on NMDA receptor function in organotypic hippocampal cultures.

Nitric oxide (NO) has been proposed to trigger long-term potentiation (LTP) at CA3 to CA1 synapses. We previously reported that NO synthesis inhibitors and blockers reduce an electrophysiological index of NMDA receptor activation in acute hippocampal slices. We now show that the NOS inhibitor, NG-methyl-L-arginine (MLA), also reversibly prevents LTP induction in organotypic hippocampal slices and significantly reduces a biochemical index of NMDA receptor function. These results results further indicate that MLA inhibits LTP induction by interfering with NMDA receptor functions.

Offline assessment of atherosclerotic coronary calcium from electron beam tomograms.

Coronary calcium screening using electron beam computed tomography (EBCT) is being applied clinically and for research purposes. We compared the accuracy of a specialized image analysis system with the standard proprietary software in the scanner's host computer. Sixty-seven symptomatic patients underwent coronary angiography and EBCT. Tomograms were analyzed using the proprietary software included in the scanner and with a specialized coronary calcium scoring work station. Sensitivities, specificities, and receiver operating characteristic curve areas were calculated for the proprietary software and the specialized system using the angiographic definition of disease of at least one stenosis causing greater than 50% luminal narrowing. There were no significant differences between the proprietary and the specialized software's accuracy. Receiver operating characteristic curve areas were 0.84 and 0.82 for proprietary software, respectively. During a 50 minute analysis session, the average number of studies analyzed were 12.6 +/- 1.7 using the proprietary software and 23.2 +/- 5.7 using the specialized software (P = .02).Image analysis was thus found to be more rapid using the specialized software. The specialized coronary calcium analysis system is as accurate as the proprietary software for scoring EBCT for coronary calcium. The reduction in analysis time makes the specialized system the preferable method.