Isolation and Characterization of Staphylococcus aureus from Raw Cow's Milk and Investigating the Effect of Bifidobacterium bifidum Probiotic Cell Free Supernatant on Their Enterotoxins Genes Expression.

The present reserach aimed to detect and isolate the genes involved in the staphylococcal enterotoxins (SEs) production in strains isolated from unprocessed cow's milk and to examine the impact of Bifidobacterium bifidum probiotic cell-free supernatant (CFS) on their expression. Standard techniques were used for isolation and identification of Staphylococci strains in unprocessed milk. The PCR was used to identify strains carrying enterotoxin genes. The B. bifidum CFS was applied to strains containing the target genes, and the genes expression levels were quantified using Real-time PCR. Using 16SrDNA sequencing, the phylogenic relationship of the isolated strains was determined. Analysis revealed that bacteria such as Staphylococcus species were found in the 72% of the samples. The PCR test showed the presence of various SE superantigens, including SEA (16.7%), SEC (11.7%), SED (8.3%), SEE (6.7%), and SEB (1.7%) in isolated strains. The B. bifidum CFS had obvious antimicrobial activity against strains 24, 51, 54, and 35 of Staphylococcus species, and the minimum inhibitory concentration and minimum bactericidal concentration values for these strains treated with B. bifidum CFS were in the range of 31.25-125 µg/ml. Strains 51 and 24 were clustered with S.aureus ATCC 25923, and strains 54 and 35 were clustered with S.aureus ATCC 12600, respectively. The RT-PCR exhibited that probiotics CFS suppressed the expression of SEA, SEB, SEC, and SEE genes (P<0.05). The average fold change for SEA, SEB, SEC, and SED genes was -1.681, -1.28, -1.52, and -0.84, respectively. The research demonstrated that probiotic bacteria can lower enterotoxin production by downregulating the expression of SEs genes.

Combined Molecular and Biochemical Identification of Alpha Toxin in Local Isolated Clostridium novyi from the Sheep Liver.

Clostridium novyi (C. novyi) causes deadly Black disease in sheep and rarely in other animals. Alpha toxin (α-toxin), the most apparent pathogen of this disease, is produced by C. novyi type B. Economic damages of C. novyi include sheep mortality costs, depreciation of affected farms, and health problems with infected carcasses. The identification of C. novyi and isolation of its pathogens by conventional methods is a time-consuming process, necessitating a simple and rapid method for isolating and detecting pathogenic C. novyi. Therefore, this study aimed to molecularly identify α-toxin in local C. novyi isolates from the sheep livers. In this study, 75 livers suspected of Black disease were sampled. The samples of the liver were cultured under anaerobic conditions. Some of the cultured colonies were used in biochemical tests. For molecular confirmation, the DNA of isolates was extracted, and the isolates were confirmed by the polymerase chain reaction (PCR) on the liver tissue and cultured samples using specific α-toxin primers. The PCR on α-toxin produced a band in the range of 609 bp, indicating that the samples belonged to C. novyi. According to the results, of 75 isolates, 18 isolates were confirmed as C. novyi. C. novyi type B was isolated from the liver and confirmed by biochemical and molecular characterization. The PCR assay ensured a sensitive and specific tool for the detection of C. novyi in the samples.

Erratum: Publisher's Note: "Laser-induced electron diffraction of the ultrafast umbrella motion in ammonia" [Struct. Dyn. 8, 014301 (2021)].

[This corrects the article DOI: 10.1063/4.0000046.].

Laser-induced electron diffraction of the ultrafast umbrella motion in ammonia.

Visualizing molecular transformations in real-time requires a structural retrieval method with Ångström spatial and femtosecond temporal atomic resolution. Imaging of hydrogen-containing molecules additionally requires an imaging method sensitive to the atomic positions of hydrogen nuclei, with most methods possessing relatively low sensitivity to hydrogen scattering. Laser-induced electron diffraction (LIED) is a table-top technique that can image ultrafast structural changes of gas-phase polyatomic molecules with sub-Ångström and femtosecond spatiotemporal resolution together with relatively high sensitivity to hydrogen scattering. Here, we image the umbrella motion of an isolated ammonia molecule (NH3) following its strong-field ionization. Upon ionization of a neutral ammonia molecule, the ammonia cation (NH3 +) undergoes an ultrafast geometrical transformation from a pyramidal ( Φ HNH = 107 ° ) to planar ( Φ HNH = 120 ° ) structure in approximately 8 femtoseconds. Using LIED, we retrieve a near-planar ( Φ HNH = 117 ± 5 ° ) field-dressed NH3 + molecular structure 7.8 - 9.8 femtoseconds after ionization. Our measured field-dressed NH3 + structure is in excellent agreement with our calculated equilibrium field-dressed structure using quantum chemical ab initio calculations.

Molecular structure retrieval directly from laboratory-frame photoelectron spectra in laser-induced electron diffraction.

Ubiquitous to most molecular scattering methods is the challenge to retrieve bond distance and angle from the scattering signals since this requires convergence of pattern matching algorithms or fitting methods. This problem is typically exacerbated when imaging larger molecules or for dynamic systems with little a priori knowledge. Here, we employ laser-induced electron diffraction (LIED) which is a powerful means to determine the precise atomic configuration of an isolated gas-phase molecule with picometre spatial and attosecond temporal precision. We introduce a simple molecular retrieval method, which is based only on the identification of critical points in the oscillating molecular interference scattering signal that is extracted directly from the laboratory-frame photoelectron spectrum. The method is compared with a Fourier-based retrieval method, and we show that both methods correctly retrieve the asymmetrically stretched and bent field-dressed configuration of the asymmetric top molecule carbonyl sulfide (OCS), which is confirmed by our quantum-classical calculations.

A case control study of Salmonella SPP. infection in stray dogs in Tehran shelters and the correlation between paraclinical tests results and clinical findings.

Salmonellosis as a zoonotic disease in dogs is not fully understood, and various reports have pointed to the transmission of antibiotic-resistant salmonella from dogs to humans. The current study aimed to evaluate the serologic and bacteriologic prevalence of Salmonella spp. in stray dogs placed in animal shelters around Tehran, compare the results to those of asymptomatic dogs, and determine the serotype of isolated species, as well as their antibiotic susceptibility pattern. A total of 100 fecal swab and blood samples were obtained from symptomatic and apparently healthy dogs (clinically) placed in four animal shelters around Tehran, Iran. Fecal and blood culture, as well as dog food culture, tube agglutination test, serotyping, and antibiotic susceptibility testing were performed on the samples. Fever, lethargy, diarrhea, and abdominal pain were observed in all the dogs in the case group, and bloody diarrhea was the least commonly detected symptom in clinical examination. A number of 11 and 4 collected fecal swabs from the case and control groups were positive for Salmonella spp., respectively. The polymerase chain reaction (PCR) also confirmed the laboratory tests results. Blood culture on the selective medium was negative for all the cases. Moreover, 60% and 100% of dogs in the case and control groups showed inflammatory markers in their blood test. The tube agglutination test was positive for 12% of the samples from the case group, while it was positive only for 5% of cases in the control group. The highest and lowest antibiotic resistance was observed against gentamicin and ciprofloxacin from the case group, respectively. Salmonella spp. infection in stray dogs placed in animal shelters is a great public health concern. In this regard, it is recommended that these animals be regularly monitored since they serve as Salmonella carriers.

Low concentrations of methamphetamine can suppress inflammation in trophoblast JEG-3 cell line induced by nicotine.

AIM: Nicotine at high concentrations induces apoptosis in trophoblastic cells through induction of cell cytotoxicity and Reactive Oxygen Species (ROS). Methamphetamine in low dose has pharmaceutical properties. It seems that this components in low dose can protect the trophoblastic cells from nicotine-induced cell death. METHOD: Trophoblastic (JEG-3) cells grown in DMEM culture medium. MTT assay test detected the cell viability and Lactate Dehydrogenase test measured the cells cytotoxicity. Griess reaction was used for NO production analysis. Cell migration traced by wounding technique. Human Cytokine Array Focused 13-plex was also used for analysis of IL-1α, IL-1ß, IL-6, INFγ, and TNFα pre-inflammatory cytokines. RESULTS: Methamphetamine, in very low dose (pM), increased the cell viability and NO production, and decreased cell cytotoxicity, IL-1α, IL-1ß, IL-6, INFγ, and TNFα pre-inflammatory cytokines of JEG-3 cell which were exposed to high dose of nicotine, respectively. Cell migration was enhanced by low dose of methamphetamine in JEG-3 cells. CONCLUSION: Methamphetamine in very low dose suppressed the JEG-3 cell death induced by high dose of nicotine (Fig. 5, Ref. 48) Keywords: methamphetamine, nicotine, cell death, NO.

Effects of low-dose morphine suppress methamphetamine-induced cell death by inhibiting the ROS generation and caspase-3 activity.

OBJECTIVE: Methamphetamine in low doses can increase vigilance and power and at high doses has destructive effects that cause toxicity and death of various cell lines and affect the central nervous system. Morphine has also protective properties, which were observed in low concentrations, for nerve cells and also seem to have the ability to reduce cell death in neural cell lines. MATERIALS AND METHODS: In this study, we used PC12 and U87 cell lines, which grew in DMEM culture media. Assays used in this study are listed below: MTT test for cell viability detection, LDH test for cytotoxicity measurement, caspase activity colorimetric assay kit (Bio-techne) for caspase 3 activity diagnosis, Rhodamine 123 for Detection of mitochondrial membrane potential. TUNNEL test for DNA fragmentation, fura-2 for Measurement of (Ca2+) ic and (Ca2+) m. fluorescence microscope for measurement of antioxidant enzyme activities. RESULTS: morphine increased cell viability and the rhodamine-123 absorbance. It reduced cell cytotoxicity, caspase 3 activity, ic (et) m Ca2+ concentration, (.OH) generation, and DNA fragmentation in all concentrations of 1 pM t0 100 nM (p < 0.05) by optimal concentration of 1 pM. CONCLUSION: morphine as a pain mediator can reduce the methamphetamine-induced cell death, may be due to its anti-inflammatory properties (Fig. 7, Ref. 52).

Low concentration of morphine protects against cell death, oxidative stress and calcium accumulation by nicotine in PC12 cells.

OBJECTIVE: Nicotine causes cell death in many cell lines. Morphine at low concentrations has protective effects against cell death. We investigated the effects of low concentration of morphine on nicotine-induced cell death in PC12 cells. MATERIALS AND METHODS: PC12 are cells that grow in DMEM culture medium. Cell viability was detected by MTT test and cells cytotoxicity was measured by LDH test. The activity of caspase-3 was diagnosed by the caspase activity colorimetric assay kit, and detection of mitochondrial membrane potential was confirmed by rhodamine 123 and TUNEL test was performed for DNA fragmentation detection. The fura-2 AM and also rhod 2-AM was used for measurement of intracellular calcium (Ca2+) ic and mitochondrial calcium (Ca2+) m and finally, measurement of antioxidant enzyme activities was assessed. RESULTS: The low concentration of morphine increased cell viability and suppressed cell cytotoxicity, cell death and the formation of mitochondrial membrane potential compared to nicotine treated cells.  It also reduced the intracellular calcium (Ca2+) ic and mitochondrial calcium (Ca2+)m concentration, respectively. CONCLUSION: Morphine as a pain reducer drug, in low concentrations, can protect PC12 cells from nicotine-induced cell death (Fig. 7, Ref. 59).

Molecular Identification of Extended-Spectrum ß-lactamase and Integron Genes in Klebsiella Pneumonia.

INTRODUCTION: Infections caused by Gram negative bacteria, producing extended-spectrum ß-lactamase, including Klebsiella pneumoniae are increasing all over the world with high morbidity and mortality. The aim of the present study was determined antimicrobial profile susceptibility and the prevalence of antibiotic resistance genes by multiplex PCR. METHODS: In the present study, we obtained one-hundred isolates of K. pneumoniae from different clinical samples. The antibiotic susceptibility testing was done in thirteen antibiotic and, therefore, M-PCRs were conducted using the DNA amplification for detection of ESBLs (blaTEM, blaCTX-M, blaSHV) and int (I, II, III) genes. RESULTS: The results of resistance to amoxicillin/clavulanate, ciprofloxacin, amikacin, trimethoprim-sulfamethoxazole, cefotaxime, ampicillin, aztreonam, imipenem, gentamicin, ceftazidime, Cefepime, ceftriaxone and levofloxacin were obtained 37%, 37%, 93%, 84%, 52%, 87%, 59%, 8%, 24%, 67%, 52%, 43% and 26%, respectively. The frequency of the extended-spectrum ß-lactamase K. pneumoniae was obtained 37%. The prevalence of resistance genes of ESBLs in the M-PCR method showed that the blaTEM, blaCTX and blaSHV were 38%, 24% and 19%, respectively, however, only 8 (8%) out of 100 isolates were found to have positive outcomes for the existence of class 1 integrons and there were no detected class 2 or class 3 integrons. CONCLUSIONS: Our results recommend the likely co-carriage of some ESBLs genes and antibiotic resistance integrons on the same plasmids harboring the MDR genes.

Three-dimensional imaging of carbonyl sulfide and ethyl iodide photodissociation using the pixel imaging mass spectrometry camera.

The Pixel Imaging Mass Spectrometry (PImMS) camera is used in proof-of-principle three-dimensional imaging experiments on the photodissociation of carbonyl sulfide and ethyl iodide at wavelengths around 230 nm and 245 nm, respectively. Coupling the PImMS camera with DC-sliced velocity-map imaging allows the complete three-dimensional Newton sphere of photofragment ions to be recorded on each laser pump-probe cycle with a timing precision of 12.5 ns, yielding velocity resolutions along the time-of-flight axis of around 6%-9% in the applications presented.

Uncultured undifferentiated adipose-derived nucleated cell fractions combined with inside-out artery graft accelerate sciatic nerve regeneration and functional recovery.

Effects of transplantation of adipose-derived nucleated cell fractions (ADNCs) on sciatic nerve regeneration were studied. A 10-mm sciatic nerve defect was bridged using artery graft filled with ADNCs. In control group, artery graft was filled with saline alone. Regenerated nerve fibres were studied for 12 weeks. In sham-operated group, sciatic nerve was only exposed and manipulated. Behavioural and functional studies confirmed faster recovery of regenerated axons in ADNCs transplanted animals than in control group (P<0.05). At the end of study period, animals in ADNCs transplanted group achieved a sciatic functional index (SFI) value of -31.6 ± -3.14, whereas in control group a value of -42.5 ± -3.7 was found. Gastrocnemius muscle mass in ADNCs transplanted animals was found to be significantly higher than that in control group (P=0.001). Morphometric indices of regenerated fibres showed the number and diameter of myelinated fibres to be significantly higher in ADNCs transplanted animals than in control group (P=0.001). On immunohistochemistry, there was more positive staining of S100 in the ADNCs transplanted animals than in control group. ADNCs transplantation into an artery graft could be considered a readily accessible technique that improves functional recovery of sciatic nerve.

ABCD2 score and BNP level in patients with TIA and cerebral stroke.

Scoring systems have been designed to help physicians in early prediction of cerebral stroke following Transitional Ischemic Attack (TIA). ABCD2 system is one of these scoring systems. Considering increase of brain natriuretic peptide following cerebral ischemic stroke, BNP level may be associated with incidence of ischemic stroke following TIA. The present study evaluates ABCD2 score, BNP level in patients with TIA and incidence of cerebral stroke. This cross sectional-analytical study evaluated 78 patients with TIA. ABCD2 score was calculated for all patients based on some criteria including age, blood pressure, clinical manifestations (speech/motor disorder), symptoms duration and diabetes. BNP level was measured at the reference laboratory when the patient referred to the treatment center. The patients were followed up for 6 months considering incidence of cerebral stroke and TIA. Mean age of the patients was 66.53 +/- 13.08 years and the sample was consisted of 62.8% male and 37.2% female patients. Mean BNP level and mean ABCD2 score was 611.31 +/- 125.61 and 4.61 +/- 10.99 in all patients, respectively. During follow-up period, TIA recurrence and cerebral stroke were, respectively seen in 11.5 and 3.8% of cases. Mortality was reported in 5.1% of the patients. BNP was significantly higher in cases with recursive TIA (p = 0.03). But, there was not any difference considering ABCD2 score (p = 0.38). BNP is capable of predicting TIA recurrence following first TIA and it can be used in this case.

The use of undifferentiated bone marrow stromal cells for sciatic nerve regeneration in rats.

In recent years, cell transplantation has become a focus of attention and reliable outcomes have been achieved in regeneration of the sciatic nerve. The effect of undifferentiated bone marrow stromal cells (BMSCs) on peripheral nerve regeneration was studied using a rat sciatic nerve regeneration model. A 10-mm sciatic nerve defect was bridged using an inside-out vein graft (IOVG) filled with undifferentiated BMSCs (2 × 10(7)cells/ml). In the control group, the vein was filled with phosphate buffer saline alone. The regenerated fibres were studied 4, 8 and 12 weeks after surgery. Assessment of nerve regeneration was based on functional (walking track analysis), histomorphometric and immunohistochemical (Schwann cell detection by S100 expression) criteria. The functional study confirmed significant recovery of regenerated axons in the IOVG/BMSC group (P<0.05). Quantitative morphometric analyses of regenerated fibres showed the number and diameter of myelinated fibres in the IOVG/BMSC group were significantly higher than in the control group (P<0.05). This demonstrates the potential for using undifferentiated BMSCs in peripheral nerve regeneration without the limitations of donor-site morbidity associated with isolation of Schwann cells. It also reduces costs because the interval between tissue collection and cell injection is reduced and the laboratory procedures are simpler compared to undifferentiated BMSCs.

Sudden death associated with myocardial contraction band necrosis in Boer goat kids.

Parenteral selenium (Se) and vitamin E (Vit E) were administered to all newborn kids at a Boer goat farm where there was previous high neonatal mortality assumed to be due to nutritional myopathy. All treated kids were affected by severe respiratory distress and died within 8 hours of Se/Vit E administration. Gross lesions included severe pulmonary edema, hydrothorax, and hydropericardium. The primary histopathologic finding was severe, acute, and monophasic myocardial contraction band necrosis. The diagnosis was accidental acute selenosis based on trace mineral analysis of the liver. This case highlights an important differential diagnosis in cases of acute myocardial contraction band necrosis and sudden death in goats and emphasizes the need for caution when administering parenteral Se/Vit E preparations.

Evaluation of pearl millet and flaxseed effects on egg production and n-3 fatty acid content.

1. A 6-week trial was conducted to evaluate the effects of pearl millet (PM) as a replacement for maize, in combination with flaxseed (FS), on productivity, egg trait parameters and egg n-3 fatty acid (FA) content in White Leghorn hens. 2. Six diet treatments were used: a control (CTL, maize-soybean meal based) diet, and diets containing 0, 2, 4, 8 or 12% FS, in which all maize was replaced by PM (PM-0, PM-2, PM-4, PM-8 and PM-12, respectively). All diets were isocaloric and isonitrogenous and met NRC (1994) requirements. Eight cage replicates per treatment were used (three hens per cage). At the end of each week, three eggs were randomly collected from each cage to measure egg trait parameters and yolks were separated, pooled and lyophilised for FA determination. Body weights and feed consumption were recorded weekly. Egg production (number of eggs and egg mass produced) was recorded daily. At the end of the experiment, all hens were killed to determine liver haemorrhage score. 3. Egg traits and flock performance parameters were not different among treatments except at week 4, when birds on the PM-12 diet produced smaller eggs than hens on the PM-0 and PM-2 diets. Yolk pigmentation scores were lower for the PM-0 and PM-2 diets (1.60 +/- 0.24 and 1.80 +/- 0.20, respectively), increased with higher inclusions of FS (2.75 +/- 0.47 for PM-12 diet) but did not reach control levels (6.00 +/- 0.01). Liver haemorrhage scores were not affected by dietary treatment. Hens given the PM-8 and PM-12 diets produced eggs with n-3 FA content higher than required to be considered as n-3 FA enriched eggs, and had a lower n-6/n-3 FA acid ratio than eggs of hens consuming CTL or the PM based diets with lower FS supplementation. 4. These results suggest that PM can be used to substitute for maize in the diets of layers and may reduce the amount of FS needed to obtain n-3 FA enriched eggs.

Absidia posttraumatic infection: successful treatment with posaconazole.

The class zygomycetes is a large group of filamentous fungi found ubiquitously in nature, primarily in humid soil. Their virulence in immunocompetent hosts is low; therefore, they are usually considered to be contaminants of cultures. In the present study, we describe a patient who had severe traumatic injury with an Absidia infection, who developed intolerance to antifungal treatment with amphotericin B, but responded to posaconazole treatment.

[Unplanned discharge in a psychiatric day clinic - argument for an individualized approach]. / Behandlungsabbruch in der psychiatrischen Tagesklinik - Plädoyer für eine differenzierte Betrachtungsweise.

OBJECTIVE: Drop-outs, respectively unplanned discharges in psychiatric therapy are often explained by so-called lack of insight into the disease or difficult circumstances of admission. We were interested in the question how often drop outs are noticed in a voluntarily day clinic setting where nearly all psychiatric diseases are treated. METHOD: The charts of all patients discharged in 1998 (n = 65) were screened referring to their mode of discharge. RESULTS: 31 patients (47.7 %) regularly ended their treatment. An unplanned discharge was registered with 34 patients (52.3 %). CONCLUSIONS: The statistically relevant term of drop-out does not appear to be the right one to deal with the individual treatment situation and the motivation for the limitation of treatment. A typology of prematurely ended therapies is developed.

Physical mapping of the srl recA region of Escherichia coli: analysis of Tn10 generated insertions and deletions.

A restriction endonuclease map for the enzymes EcoRI, BamHI, SalI, and PstI covering 23.5 kilobase pairs (kb) of the srl recA region of Escherichia coli was constructed. An insertion of the transposon Tn10 in the negative regulatory gene srlR was shown to be located 5.8 kb away from the promoter proximal end of the recA gene. The extent of several Tn10 generated deletions, originating from the srlR301::Tn10 insertion, were analyzed by physical mapping. Three mutations that had removed the Tn10 encoded tetracycline resistance gene, del(srl-recA)302, del(srl-recA)304, and del(srl-recA)303, were found to be deleted for 40%, 45%, and 50% of the recA structural gene, respectively. A deletion, del(srl-recA)306, that had not affected the structure of the Tn10 in srlR301 was shown to have removed the entire recA structural gene.