Characterization of resistance to tetracyclines and aminoglycosides of sheep mastitis pathogens: study of the effect of gene content on resistance.

AIMS: Mastitis causes economic losses and antimicrobials are frequently used for mastitis treatment. Antimicrobial resistance surveys are still rare in the ovine field and characterization of strains is important in order to acquire information about resistance and for optimization of therapy. METHODS AND RESULTS: Bacterial pathogens recovered in milk samples from mastitis-affected ewes were characterized for resistance to tetracyclines and aminoglycosides, members of which are frequently used antimicrobials in small ruminants. A total of 185 strains of staphylococci, streptococci, and enterococci, common mastitis pathogens, were tested for minimal inhibitory concentration (MIC) to tetracycline, doxycycline, minocycline, gentamicin, kanamycin, streptomycin, and for resistance genes by PCR. Effects of different tet genes arrangements on MICs were also investigated. Staphylococci expressed the lowest MIC for tetracycline and tet(K) was the most common gene recovered; tet(M) and tet(O) were also found. Gene content was shown to influence the tetracycline MIC values. Enterococci and streptococci showed higher MICs to tetracyclines and nonsusceptible strains always harboured at least one ribosomal protection gene (MIC above 8 µg ml(-1) ). Streptococci often harboured two or more tet determinants. As regards the resistance to aminoglycosides, staphylococci showed the lowest gentamicin and kanamycin median MIC along with streptomycin high level resistant (HLR) strains (MIC >1024 µg ml(-1) ) all harbouring str gene. The resistance determinant aac(6')-Ie-aph(2″)-Ia was present in few strains. Streptococci were basically nonsusceptible to aminoglycosides but neither HLR isolates nor resistance genes were detected. Enterococci revealed the highest MICs for gentamicin; two str harbouring isolates were shown to be HLR to streptomycin. CONCLUSION: Evidence was obtained for the circulation of antimicrobial-resistant strains and genes in sheep dairy farming. SIGNIFICANCE AND IMPACT OF THE STUDY: Tetracycline MIC of 64 µg ml(-1) and high-level resistance were detected for streptomycin (MIC >1024 µg ml(-1) ), so that effectiveness of common treatments may be at risk.

Label-free live characterization of mesenchymal stem cell spheroids by biophysical properties measurement.

Three-dimensional (3D) cell culture has become a consolidated method in the stem cell field, where mesenchymal stromal stem cells (MSCs) can be used to generate in vitro spheroid aggregates called MSC-Spheroids (MSph). MSph is a floating cluster of stem cells similar to those in literature are known as bone marrow-derived "mesenspheres". Even though MSC properties are shared by MSph, depending on the cell type and their tissue source, the morphology and degree of compaction of the MSph can be variable, creating limitations in such a cell model. Thus, during culture, a variation in stem cell functionality and viability, in addition to the suitability for comparing MSph in some experimental protocols, can be affected by spheroid biophysical intrinsic properties like mass density. To investigate this limitation and provide a new method for researchers, MSph of seven different tissue sources were compared by combining mass density, weight, and size evaluations with viability assays for ATP measurement. MSph cultured in traditional static conditions showed decreased in viability over the days of culture, while mass density exhibited different trends among cell types. Additionally, treatment of MSph with a non-toxic concentration of a natural compound cell regulator, such as plumbagin, altered the mass density of a selected cell type, thereby confirming the efficacy of the biophysical approach in monitoring MSph variability post-treatment. The results encourage using MSph in the early days of culture after their formation to ensure viability and likely retention of the stem cell phenotype.

Patients with cancer who will be cured and projections of complete prevalence in Italy from 2018 to 2030.

BACKGROUND: The number and projections of cancer survivors are necessary to meet the healthcare needs of patients, while data on cure prevalence, that is, the percentage of patients who will not die of cancer by time since diagnosis, are lacking. MATERIALS AND METHODS: Data from Italian cancer registries (duration of registration ranged from 9 to 40 years, with a median of 22 years) covering 47% of the population were used to calculate the limited-duration prevalence, the complete prevalence in 2018, projections to 2030, and cure prevalence, by cancer type, sex, age, and time since diagnosis. RESULTS: A total of 3 347 809 people were alive in Italy in 2018 after a cancer diagnosis, corresponding to 5.6% of the resident population. They will increase by 1.5% per year to 4 012 376 in 2030, corresponding to 6.9% of the resident population, 7.6% of women and ∼22% after age 75 years. In 2030, more than one-half of all prevalent cases (2 million) will have been diagnosed by ≥10 years. Those with breast (1.05 million), prostate (0.56 million), or colorectal cancers (0.47 million) will be 52% of all prevalent patients. Cure prevalence was 86% for all patients alive in 2018 (87% for patients with breast cancer and 99% for patients with thyroid or testicular cancer), increasing with time since diagnosis to 93% for patients alive after 5 years and 96% after 10 years. Among patients who survived at least 5 years, the excess risk of death (1 - cure prevalence) was <5% for patients with most cancer types except for those with cancers of the breast (8.3%), lung (11.1%), kidney (13.2%), and bladder (15.5%). CONCLUSIONS: Study findings encourage the implementation of evidence-based policies aimed at improving long-term clinical follow-up and rehabilitation of people living after cancer diagnosis throughout the course of the disease. Updated estimates of complete prevalence are important to enhance data-driven cancer control planning.

Visualizing breast cancer using the Twente photoacoustic mammoscope: what do we learn from twelve new patient measurements?

We acquired images of breast malignancies using the Twente photoacoustic mammoscope (PAM), to obtain more information about the clinical feasibility and limitations of photoacoustic mammography. Results were compared with conventional imaging and histopathology. Ten technically acceptable measurements on patients with malignancies and two measurements on patients with cysts were performed. In the reconstructed volumes of all ten malignant lesions, a confined region with high contrast with respect to the background could be seen. In all malignant cases, the PA contrast of the abnormality was higher than the contrast on x-ray mammography. The PA contrast appeared to be independent of the mammographically estimated breast density and was absent in the case of cysts. Technological improvements to the instrument and further studies on less suspicious lesions are planned to further investigate the potential of PAM.

Paraneoplastic Acanthosis Nigricans: The importance of exhaustive and repeated malignancy screening.

Paraneoplastic acanthosis nigricans (P-AN) characteristically has a sudden onset, rapid progression, and extensive cutaneous involvement. The association between P-AN and internal malignancy is well established and the most common association is with adenocarcinoma of gastrointestinal origin. We present the case of an 81-year-old man with a 12-month history of anorexia, weight loss, and clinical evidence of extensive acanthosis nigricans. After exhaustive and repeated investigations a papillary thyroid carcinoma and a follicular adenoma were identified and he improved upon its resection. To our knowledge, P-AN in association with thyroid neoplasm has been reported on only one previous occasion.

Optimization of acoustic coupling for bottom actuated scattering based subsurface scanning probe microscopy.

The characterization of buried nanoscale structures nondestructively is an important challenge in a number of applications, such as defect detection and metrology in the semiconductor industry. A promising technique is Subsurface Scanning Probe Microscopy (SSPM), which combines ultrasound with Atomic Force Microscopy (AFM). Initially, SSPM was used to measure the viscoelastic contrast between a subsurface feature and its surrounding medium. However, by increasing the ultrasonic frequency to >1 GHz, it has been shown that SSPM can also measure acoustic impedance based contrasts. At these frequencies, it becomes difficult to reliably couple the sound into the sample such that the AFM is able to pick up the scattered sound field. The cause is the existence of strong acoustic resonances in the sample, the transducer, and the coupling layer-the liquid layer used to couple the sound energy from the transducer into the sample-in combination with the nonlinearity of the tip-sample interaction. Thus, it is essential to control and measure the thickness of the coupling layer with nanometer accuracy. Here, we present the design of a mechanical clamp to ensure a stable acoustic coupling. Moreover, an acoustic method is presented to measure the coupling layer thickness in real-time. Stable coupling layers with thicknesses of 700 ± 2 nm were achieved over periods of 2-4 h. Measurements of the downmixed AFM signals showed stable signal intensities for >1 h. The clamp and monitoring method introduced here makes scattering based SSPM practical, robust, and reliable and enables measurement periods of hours.

A systematic review of the effects of adding neostigmine to local anesthetics for neuraxial administration in obstetric anesthesia and analgesia.

BACKGROUND: Drugs used in obstetric patients must accomplish two goals: efficacy and safety for both mother and fetus. Neostigmine has been co-administered epidurally and intrathecally with local anesthetics and other adjuncts in the obstetric setting. The aim of this meta-analysis was to assess the efficacy and incidence of adverse events related to the use of neostigmine in obstetric anesthesia. METHODS: A meta-analysis of randomized-controlled human trials was conducted using the data sources Google Scholar and PubMed (updated 1 November 2014). Inclusion criteria were: random allocation to treatment; comparison of neostigmine or neostigmine with local anesthetics and/or other adjuvants versus placebo or placebo with local anesthetics and/or other adjuvants; and approval by an ethics committee. RESULTS: The use of neostigmine as an adjuvant in neuraxial anesthesia is associated with a reduction in the dose of local anesthetic during labor analgesia and postoperative analgesia following cesarean section: mean reduction of local anesthetic (ropivacaine or bupivacaine) vs. control -4.08 (95% CI -6.7 to -1.5) mg/h (P=0.002). The risk of nausea was increased vs. control with intrathecal neostigmine (OR 8.99 [95% CI 4.74 to 17.05], P <0.001) but not with epidural neostigmine (OR 0.97 [95% CI 0.46 to 2.05], P=0.94). Use of neuraxial neostigmine was associated with a decrease in the risk of pruritus but there was no increase in the incidence of hypotension, dizziness or sedation and no effect on the incidence of abnormal fetal heart rate patterns or Apgar scores. CONCLUSIONS: Neuraxial administration of neostigmine significantly reduces local anesthetic consumption without serious adverse side effects to the mother or fetus. However, neostigmine is only recommended for epidural administration as intrathecal use significantly increases the incidence of maternal nausea and vomiting.

Photoacoustic image patterns of breast carcinoma and comparisons with Magnetic Resonance Imaging and vascular stained histopathology.

Photoacoustic (optoacoustic) imaging can visualize vasculature deep in tissue using the high contrast of hemoglobin to light, with the high-resolution possible with ultrasound detection. Since angiogenesis, one of the hallmarks of cancer, leads to increased vascularity, photoacoustics holds promise in imaging breast cancer as shown in proof-of-principle studies. Here for the first time, we investigate if there are specific photoacoustic appearances of breast malignancies which can be related to the tumor vascularity, using an upgraded research imaging system, the Twente Photoacoustic Mammoscope. In addition to comparisons with x-ray and ultrasound images, in subsets of cases the photoacoustic images were compared with MR images, and with vascular staining in histopathology. We were able to identify lesions in suspect breasts at the expected locations in 28 of 29 cases. We discovered generally three types of photoacoustic appearances reminiscent of contrast enhancement types reported in MR imaging of breast malignancies, and first insights were gained into the relationship with tumor vascularity.

Efficacy of nicardipine slow release (SR) on hypertension, potassium balance and plasma aldosterone in idiopathic aldosteronism.

We evaluated the efficacy and tolerance of a dihydropyridine calcium antagonist, nicardipine slow release, in eight patients with idiopathic aldosternism. Nicardipine (80 mg/day) was given orally for 12 weeks and no dietary restrictions were imposed. During the study measurements were made of supine blood pressure, plasma renin activity, plasma aldosterone concentration, and serum potassium. Nicardipine lowered systolic and diastolic blood pressure to normal, plasma aldosterone was reduced and serum potassium levels were increased. Basal renin concentration was not altered by nicardipine. There were no side effects sufficient to cause withdrawal from the study. These results suggest that nicardipine, for efficacy and tolerance, may represent an alternative among Ca2+ channel blockers, either controlling blood pressure or reducing aldosterone levels in patients with idiopathic aldosteronism.

[Acute bilateral sensorineural hearing loss caused by immuno-mediated inner ear disease]. / Sordera neurosensorial aguda bilateral por enfermedad del oído interno inmuno-mediada.

The audiovestibular system can be affected by an immunologic etiology. The immune-mediated inner ear disease (IMIED) is a syndrome that includes rapidly progressive sensorineural hearing loss, vertigo and tinnitus, which occurs as a primary disorder or complicates certain autoimmune systemic conditions. However, if treated promptly with immunosuppression, the audiological sequel of IMIED may be avoided. We present a 28 year old female patient, who after rhinitis and mioarthralgias developed a vestibular syndrome. A week later she experienced bilateral hearing loss that progressed to deafness in 72 hours. The examination revealed horizontal and torsional nystagmus, a disrupted vestibulo-ocular reflex and vertigo with the positional changes. Laboratory data were normal except for eritrosedimentation rate (75 mm/1 hour). The autoantibodies usually present in rheumatologic autoimmune systemic diseases were negative. The antibodies to the 68-kD antigen found in the inner ear were positive. The chest x-ray and sinus x-ray were normal. The head magnetic resonance imaging with gadolinium and ear computed tomography were normal. Cerebrospinal fluid studies showed normal findings. With the possible diagnosis of IMIED we started early treatment with corticosteroids, with improvement in auditory and vestibular function thereafter. We highlight the early recognition of IMIED as a differential diagnosis in patients with acute bilateral hearing loss, because prompt treatment with immunosuppression might have a positive effect on auditory function recovery.

[Idiopathic osteosclerosis in jaw bones. Clinical statistic study on a population of Sardinian origin]. / Osteosclerosi idiopatiche dei mascellari. Indagine clinico-statistica su una popolazione originaria della Sardegna.

BACKGROUND: Idiopathic Osteosclerosis are localized, non-expansive and asymptomatic areas of bone radio-opacity, whose aetiology is unknown. The aim of this study was to assess its frequency, and distribution according to its location and to patients' age and gender, in a sample of patients native in Sardinia. METHODS: 697 consecutive clinical records of patients who underwent odontological treatment at the Department of Odontostomatologic Sciences at Cagliari University, over a 10-year period were studied. The panoramic radiographs attached to the clinical records were examined and searched for all cases that showed radio-opacity areas in maxillary bones that could be identified as idiopathic osteosclerosis (IO). RESULTS: By examining orthopantomographies we have located 58 IOs. We found no significant differences between the sexes; average age was 37.8 years. The mandible has proved to be the most affected site (98.2%) in the first molar tooth region (34.5%), followed by the premolar teeth region (25.9%) and of the second molar tooth (15.5%). The lesions examined had an average diameter of 8.8 mm. CONCLUSIONS: IO frequency has proved to be slightly higher than that reported in other articles, though remaining significantly lower than the values reported in Eastern populations. From the clinical point of view, IOs have been identified as sporadic radiographic findings without any real pathological relevance, except for one case in which the lesion caused nervous compression and dental retention, so that it was necessary to administer specific treatment.

Role of angiogenesis inhibitors in colorectal cancer: sensitive and insensitive tumors.

Angiogenesis is a key factor in the carcinogenesis process. In oncological practice, angiogenesis inhibition, mainly through the blockade of the VEGF family and its receptors, has been robustly demonstrated to produce clinical benefits and, in specific disease subsets such as colorectal cancer, to extend the overall survival of treated patients. VEGF is a multifunctional growth factor that mediates its functions through cognate receptors on endothelial cells and it has been discovered for its capability to induce macromolecule hyperpermeability in veins and venules. Several approaches have been taken to target angiogenesis in cancer: drugs that target one or more soluble ligands of the VEGF family, drugs that selectively inhibit one or more receptors of the VEGF receptor family, and drugs that inhibit VEGF receptor(s) among other, non VEGF-related targets. At present, two compounds have shown significant clinical activity, bevacizumab, Avastin® and aflibercept, Zaltrap®, and only one of these (bevacizumab) has so far been registered for use in clinical practice. In the present review, we explore and summarize the main features of the angiogenetic process, concerning in particular a common and potentially lethal disease as colorectal cancer. We overview the molecular pathways that characterize angiogenesis, focusing on VEGF family, the current applications and limitations of its blockade in oncology, and the hypothetical future perspectives of anti-angiogenic therapy.

[Indications and limits to endourologic procedures for endometriosis of the urinary tract]. / Indicazioni e limiti delle procedure endourologiche nell'endometriosi dell'apparato urinario.

INTRODUCTION: The incidence of Urinary tract endometriosis (UTE) ranges from 1% to 3%; bladder is the most affected organ (85% of UTE), followed by ureter (12 - 14% of UTE), for which we distinguish an intrinsic very rare form and an extrinsic variety most frequently occurring in advanced pelvic endometriosis. MATERIALS AND METHODS: From 1997 to 2010, 33 surgical procedures for urologic endometriosis were performed, involving the urinary tract, in 28 patients with mean age of 31 years (25-43). The localization of endometriosis were: 7 cases in the bladder, 2 cases in the vesicoureteral tract, and 19 cases of ureteral tract only. Of these, two cases were diagnosed with an intrinsic localization. RESULTS: Overall, we performed 3 TURB, 5 partial cystectomies (2 with open surgical approach and 3 by laparoscopy procedure), 12 laparoscopic ureterolysis and simultaneous protection of the upper urinary tract with stent, 9 cases of ureterocystoneostomy (UCNS) according to Lich-Gregoire procedure, and 3 according to Boari-Kuess procedure. Of the 12 patients who underwent ureterolysis with laparoscopic and stenting procedure, five cases required a UCNS according to Lich-Gregoire technique for persistent ureteral obstruction. CONCLUSIONS: The limits of endoscopic procedures in endometriosis of the urinary tract are correlated both to the degree of extension and the localization of the disease. It is mandatory to achieve an interdisciplinary consensus in order to ensure the disease removal and the simultaneous functional results of the upper urinary tract.

[Three cases of renal cell carcinoma with sarcomatoid differentiation]. / Tre casi di carcinoma renale con differenziazione sarcomatoide.

Renal cell carcinoma with sarcomatoid differentiation has an incidence of 4-6%. It occurs more frequently with clinical advanced presentation in relation to the greater biological aggressiveness. This variant was also found both in transplanted kidneys and in patients with von Hippel-Lindau syndrome. The authors present three cases of cancer all clinically evolving to rapidly progressive conditions. The pathologic staging was, respectively: Case 1 pT4 N0 M0 G4 case, Case 2 P G4 T3a N2 M0, Case 3 pT3a N0 M1 G4. The clinical responses related to protocols with VEGF drugs that seem to have better clinical response compared to immunotherapy are still being studied. From the pathologic point of view it is necessary, for all renal cell cancers, to search and identify the sarcomatoid components that, although poorly represented, give a negative prognosis.

[A case of chronic schistosomiasis four years after infestation]. / Caso di schistosomiasi cronica a quattro anni dalla infestazione.

Authors present a case of bilharziosis incidentally diagnosed in a patient undergoing TURB for suspected bladder cancer. The patient, who in 2005 had gone to Malaysia, had been suffering from recurrent hemorrhagic cystitis since 2007, which were treated with antibiotic therapy. In November 2009 he presented to our observation for persistent hematuria, underwent ultrasound examination, fibroscopy and TURB diagnostics for suspicious lesions. The histopathology diagnosis found granulomatous lesions with typical parasites eggs due to schistosomiasis eggs. As a consequence of that, the patient underwent medical therapy. The pathologist's role becomes nullifying not only for the diagnosis of parasitic infections but also for the exclusion or evidence of urothelial squamous neoplasia. The low incidence of this rare parasitic disease in European tourists and the presence of immigrants in our country require to spread the knowledge of these parasites and the most simple tests for early detection.

Bitter taste study in a sardinian genetic isolate supports the association of phenylthiocarbamide sensitivity to the TAS2R38 bitter receptor gene.

Recently, a major locus on chromosome 7q was found in association with the taste sensitivity to phenylthiocarbamide (PTC) in humans. This region contains the TAS2R38 gene that encodes a member of the TAS2R bitter taste receptor family. Three SNPs within this gene demonstrated a strong association with taster status in Utah families and in an additional sample of 85 unrelated individuals. We studied a small isolated village in eastern Sardinia and carried out a genome-wide scan to map the genetic basis of PTC perception in this population. We performed both qualitative and quantitative PTC-taste linkage analysis. Qualitative analysis was carried out by defining a cut-off from the bimodal distribution of the trait and classifying subjects as tasters and non-tasters (75 and 25%, respectively). Linkage analysis on 131 subjects belonging to a unique large multi-generation pedigree comprising 239 subjects confirmed significant evidence for linkage at 7q35 also in our population. Haplotype analyses of the three SNPs inside the PTC gene allowed us to identify only two haplotypes that were associated with the non-taster phenotype (80% AVI homozygous) and to taster phenotype (40% PAV homozygous and 56% PAV/AVI heterozygous). Sex, age and haplotype effect explained 77.2 % of the total variance in PTC sensitivity.