Anammox bacteria enrichment and characterization from municipal activated sludge.

A sustainable option for nitrogen removal is the anaerobic ammonium-oxidizing (anammox) process in which ammonium is oxidized to nitrogen gas with nitrite as electron acceptor. Application of this process, however, is limited by the availability of anammox biomass. In this study, two Brocadia-like anammox phylotypes were successfully enriched, detected and identified from an activated sludge taken from a domestic wastewater treatment plant (Minas Gerais, Brazil) employing a Sequencing Batch Reactor (SBR). The dominant phylotype was closely related to 'Candidatus Brocadia sinica', but one clone seemed to represent a novel species for which we propose the name 'Candidatus Brocadia brasiliensis'. Based on Fluorescence in situ hybridization (FISH) analysis, this enrichment led to a relative population size of 52.7% (±15.6) anammox bacteria after 6 months of cultivation. The cultivation process can be divided into three phases: phase 1 (approximately 25 days) was characterized by heterotrophic denitrification metabolism, phase 2 was the propagation phase and phase 3 (from the 87th day onwards), in which significant anammox activity was detected. A long-term performance of the SBR showed a near perfect removal of nitrite based on the influent NO(2)(-)-N concentration of 61-95 mg L(-1). The average ammonia removal efficiency was 90% with the influent NH(4)(+)-N concentration of 55-82 mg L(-1). Therefore, anammox cultivation and enrichment from activated sludge was possible under a controlled environment within 3 months.

The Canadian Neuromuscular Disease Registry 2010-2019: A Decade of Facilitating Clinical Research Througha Nationwide, Pan-NeuromuscularDisease Registry.

We report the recruitment activities and outcomes of a multi-disease neuromuscular patient registry in Canada. The Canadian Neuromuscular Disease Registry (CNDR) registers individuals across Canada with a confirmed diagnosis of a neuromuscular disease. Diagnosis and contact information are collected across all diseases and detailed prospective data is collected for 5 specific diseases: Amyotrophic Lateral Sclerosis (ALS), Duchenne Muscular Dystrophy (DMD), Myotonic Dystrophy (DM), Limb Girdle Muscular Dystrophy (LGMD), and Spinal Muscular Atrophy (SMA). Since 2010, the CNDR has registered 4306 patients (1154 pediatric and 3148 adult) with 91 different neuromuscular diagnoses and has facilitated 125 projects (73 academic, 3 not-for-profit, 3 government, and 46 commercial) using registry data. In conclusion, the CNDR is an effective and productive pan-neuromuscular registry that has successfully facilitated a substantial number of studies over the past 10 years.

Dynamics of Aeromonas species isolated from wastewater treatment system.

Aeromonas are widely distributed in the aquatic environment, and are considered to be emerging organisms that can produce a series of virulence factors. The present study was carried out in a sanitary sewage stabilization pond treatment system, located in Lins, State of São Paulo, Brazil. Most probable number was applied for estimation of the genus Aeromonas. Colony isolation was carried out on blood agar ampicillin and confirmed by biochemical characterization. Aeromonas species were isolated in 72.4% of influent samples, and in 55.2 and 48.3% of effluent from anaerobic and facultative lagoons, respectively. Thirteen Aeromonas species were isolated, representing most of the recognized species of these organisms. Even though it was possible to observe a tendency of decrease, total elimination of these organisms from the studied system was not achieved. Understanding of the pathogenic organism's dynamics in wastewater treatment systems with a reuse potential is especially important because of the risk it represents.

Molecular detection of enterotoxins in environmental strains of Aeromonas hydrophila and Aeromonas jandaei.

Aeromonas species are widely distributed in aquatic environments and recent studies include the genus in the emergent pathogens group because of its frequent association with local and systemic infections in immunocompetent humans. Aiming to search for virulence genes in environmental strains of Aeromonas hydrophila and Aeromonas jandaei, we designed specific primers to detect act/hlyA/aer complex and alt genes. Primers described elsewhere were used to detect ast. Eighty-seven strains previously identified using phenotypic and genotypic tests as A. hydrophila (41) and A. jandaei (46) were analysed for the presence of the virulence genes using PCR. DNA fragments of expected size were purified and directly sequenced. Among the 41 strains of A. hydrophila 70.7% (29), 97.6% (40) and 26.8% (11) possessed act/hlyA/aer complex, ast and alt genes, respectively. Among the 46 strains of A. jandaei, 4.4% (2), 0% (0) and 32.6% (15) were positive for act/hly A/aer complex, ast and alt genes, respectively. Sequencing allowed for the confirmation of amplified products using BLAST. The present work proposes a specific and rapid diagnostic method to detect the main virulence determinants of Aeromonas, a genus potentially pathogenic to humans.

Detection of metallo-beta-lactamases-encoding genes in environmental isolates of Aeromonas hydrophila and Aeromonas jandaei.

AIMS: To determine the prevalence and expression of metallo-beta-lactamases (MBL)-encoding genes in Aeromonas species recovered from natural water reservoirs in southeastern Brazil. METHODS AND RESULTS: Eighty-seven Aeromonas isolates belonging to Aeromonas hydrophila (n = 41) and Aer. jandaei (n = 46) species were tested for MBL production by the combined disk test using imipenem and meropenem disks as substrates and EDTA or thioglycolic acid as inhibitors. The presence of MBL genes was investigated by PCR and sequencing using new consensus primer pairs designed in this study. The cphA gene was found in 97.6% and 100% of Aer. hydrophila and Aer. jandaei isolates, respectively, whereas the acquired MBL genes bla(IMP), bla(VIM) and bla(SPM-1) were not detected. On the other hand, production of MBL activity was detectable in 87.8% and 10.9% of the cphA-positive Aer. hydrophila and Aer. jandaei isolates respectively. CONCLUSIONS: Our results indicate that cphA seems to be intrinsic in the environmental isolates of Aer. hydrophila and Aer. jandaei in southeastern Brazil, although, based on the combined disk test, not all of them are apparently able to express the enzymatic activity. SIGNIFICANCE AND IMPACT OF THE STUDY: These data confirm the presence of MBL-producing Aeromonas species in natural water reservoirs. Risk of waterborne diseases owing to domestic and industrial uses of freshwater should be re-examined from the increase of bacterial resistance point of view.

Sanitation assessment of wastewater treated by stabilization ponds for potential reuse in agricultural irrigation sanitation assessment.

Wastewater reuse has become an important alternative to agricultural irrigation; on the other hand, it poses concern with regard to public health. Total coliform and Escherichia coli concentration, presence of helminth eggs and Salmonella, and physical-chemical parameters were evaluated in raw and treated wastewater. Chemical and biochemical oxygen demand removal efficiency was 74.6 and 77.9%, respectively. As for organic nitrogen, total phosphorus, and total suspended solids, total efficiency removal was 17.4, 12.5, and 32.9%, respectively. The average density of total coliforms and E. coli was 3.5 x 10(9) and 1.8 x 10(8) MPN/100 mL and 1.1 x 10(7) MPN/100 mL and 3.9 x 10(5) MPN/100 mL for raw and treated wastewater, respectively. Ascaris eggs were observed in 80.8% of the samples collected, and viable eggs in 42.3% of the samples. Salmonella was detected in 36.4% of the samples. The values observed in treated wastewater did not show the adequate bacteriological quality, as recommended by World Health Organization (Geneva, Switzerland). Therefore, additional measures should be taken to achieve an improved microbiological and parasitological quality.

High-affinity chimeric anti-(colorectal carcinoma) antibody correlated to enhanced tumor targeting in biodistribution and imaging.

The genetically engineered chimeric cB72.3m4 and cB72.3m12 antibodies recognize the same tumor-associated TAG72 antigen. The high-affinity cB72.3m4 antibody had an approximately 18-fold higher affinity constant for the TAG72 antigen than the low-affinity cB72.3m12 antibody. The relationship between antibody affinity and tumor targeting was studied by using these two antibodies. In biodistribution and imaging studies in athymic mice bearing LS174T human colon cancer xenografts, the radiolabelled high-affinity cB72.3m4 antibody was rapidly cleared from the blood, whereas the low-affinity cB72.3m12 antibody had slower blood clearance. The data showed that the high-affinity cB72.3m4 antibody appeared to localize more in tumors (based on tumor:normal-tissue ratios) than did the low-affinity cB72.3m12 antibody, and enhanced the target-to-nontarget image contrast. This study provides evidence that the high-affinity chimeric antibody cB72.3m4 may be useful in both immunodetection and immunotherapy of cancer.

Potassium-channel opener cardioplegia is superior to St. Thomas' solution in the intact animal.

BACKGROUND: In isolated hearts, the potassium-channel opener pinacidil is an effective cardioplegic agent. This study tested the hypothesis that pinacidil is superior to St. Thomas' solution in the more clinically relevant intact animal. METHODS: Sixteen pigs were placed on full cardiopulmonary bypass. Hearts underwent 2 hours of global ischemia (10 degrees to 15 degrees C). Either St. Thomas' or 100 micromol/L pinacidil was administered every 20 minutes (10 mL/kg). Preischemic and postreperfusion slopes of the preload-recruitable stroke work relationship were determined. Changes in myocardial adenine nucleotide levels and cellular ultrastructure were analyzed. RESULTS: Pinacidil cardioplegia resulted in an insignificant change in the slope of the preload-recruitable stroke work relationship (40.6+/-2.1 mm Hg/mm before ischemia and 36.5+/-3.7 mm Hg/mm after ischemia; p = 0.466). In contrast, St. Thomas' solution resulted in a significant decrease in the slope after reperfusion (34.3+/-5.5 mm Hg/mm and 13.5+/-2.3 mm Hg/mm; p = 0.003). Adenine nucleotide levels, myocardial tissue water, and ultrastructural changes were similar between groups. CONCLUSIONS: Pinacidil ameliorated myocardial stunning associated with traditional hyperkalemic cardioplegia without causing significant differences in cellular metabolism.

Biological evaluation of 2-fluoro-2-[123I]iodo-mannose (FIM): biological evaluation of FIM.

This paper describes the biodistribution of a radio-iodinated analog of fluorodeoxyglucose (FDG). 123I-2-fluoro-2-iodo-mannose (FIM) was investigated as a potential single photon emission tomography (SPECT) imaging agent. We also compare the results with the observed distribution of the classical PET agent 18F-FDG and newly developed 18F-difluorodeoxyglucose (DFDG). Following radioiodination, the final product was stable in-vitro for 24 hrs. Mice showed a rapid blood clearance and deiodination of the 123I-FIM reflected by high stomach and thyroid uptake. Comparison with 18F-FDG and 18F-DFDG revealed a large discrepancy between the 18F labeled sugars and the 123I-FIM biological distribution. The iodinated product was not found to be a metabolic marker for in-vivo studies.

Immunoscintigraphic detection of primary and metastatic spontaneous canine osteosarcoma with F(ab')2 fragments of osteosarcoma-associated monoclonal antibody TP-1.

Monoclonal antibody TP-1 has been shown to bind selectively to human and canine osteosarcoma cells in vitro using immunohistochemical stains. This report describes the in vivo administration of radioiodinated F(ab')2 fragments of monoclonal antibody TP-1 in dogs with primary and/or metastatic spontaneous osteosarcoma. Two dogs were injected with 131labeled F(ab')2 TP-1 and two dogs were injected with 123labeled antibody fragments. Immunoscintigraphy successfully demonstrated the radiolabeled antibody fragments in 6/6 known primary or metastatic lesions and in addition detected 4 metastatic lesions not diagnosed by conventional radiographs. Concurrent imaging of 99mTc labeled autologous erythrocytes in two dogs confirmed that the accumulation of radiolabeled antibody fragments was independent of the blood pool. There was no immunoscintigraphic evidence of localization of radioiodine to normal organs or tissues other than those expected to accumulate free iodine. The present study has demonstrated the potential of monoclonal antibody TP-1 F(ab')2 fragments for early detection of metastatic spread of spontaneous osteosarcoma.

Establishment of a rat colonic carcinoma model for study of immunoreagents against the human tumor-associated TAG72 antigen.

TAG72 originally defined by the mouse B72.3 antibody is a mucin-like, human tumor-associated antigen present in more than 85% of human colonic adenocarcinomas. Establishment of a tumor model expressing the TAG72 antigen in immunocompetent animals would be of great benefit in evaluating the therapeutic efficacy and studying anti-tumor immune mechanisms of anti-TAG72 immunoreagents. In this study, we screened 6 animal tumor cell lines including 3 derived from mouse colonic adenocarcinomas (MCR-26, MCR-38-LD and CA-51), 1 from mouse ovarian adenocarcinoma (MOT), 1 from rat colonic adenocarcinoma LMCR, and 1 from rat mammalial adenocarcinoma (R3230AB) for TAG72 expression by using the B72.3 antibody. Immunohistochemistry disclosed significant amounts of TAG72-expression in the dimethylhydrazine-induced BDIX rat colonic adenocarcinoma LMCR. The rat TAG72 antigen purified from rat LMCR tumors showed strong immunoreactivity for the B72.3 antibody in ELISA analysis and displayed a smear band of high molecular weight in Western blotting, which is similar to the human TAG72 antigen purified from human LS174T colonic adenocarcinoma. In addition, the iodinated B72.3 antibody was able to localize LMCR tumor in BDIX rats. Therefore, this rat LMCR model should be useful in studying human colonic cancer, especially in evaluating the therapeutic efficacy of anti-TAG72 immunoreagents such as the recombinant fusion proteins possessing the anti-TAG72 antibody fragment and the cytokine moiety.

Pharmaceutical interference with the [14C] carbon urea breath test for the detection of Helicobacter pylori infection.

Helicobacter pylori bacteria reside in the mucosal lining of the stomach where, due to a variety of factors, the infection predisposes patients to peptic ulcer disease. Detection of H. pylori is important in the treatment and follow-up of patients with peptic ulcer disease and the urea breath test is the method of choice. This article will briefly review the methods for diagnosing H. pylori, emphasizing the [(14)C]urea breath test. The agents which can interfere with the results of the breath test will be reviewed and the role of the consulting pharmacist will be discussed.

Pharmacologic interventions in nuclear medicine assessment of cardiac perfusion.

Drugs that are currently used for therapeutic purposes can also be used in diagnostic tests. This paper will review the use of such pharmacological interventions in cardiac assessment in Nuclear Medicine. To fully comprehend the effect of these drugs, a small review of diagnostic nuclear medicine as currently used to assess cardiac perfusion is included. This will allow pharmacists to understand the rationale behind the single administration of either vasodilator or inotropic agents and to review which drugs and food may interact with the test.

Immunohistochemical staining and radionuclide imaging of canine tumors, using a monoclonal antibody recognizing a synthetic carbohydrate antigen.

The in vitro and in vivo binding of a monoclonal antibody (MAB) that recognizes a tumor-associated carbohydrate antigen was studied in dogs. Monoclonal antibody 155H.7 was raised in response to innoculation of mice with beta-galactose(1-3)beta N-acetylgalactosamine conjugated to human serum albumin. Avidin-biotin-complex immunohistochemical staining of cryostat sections of normal and neoplastic canine tissue specimens revealed heterogenous binding of MAB 155H.7 to the cells of many canine mammary and lung carcinomas and homogenous staining of many sarcomas, including osteogenic sarcoma. In addition, there was variable staining of a variety of normal tissues including some ductual epithelium, peripheral nerve fibers, and some endothelial cells and fibroblasts. Immunoscintigraphy with 131I-labeled MAB 155H.7 was used to study the in vivo distribution of the antibody. The 131I-labeled MAB 155H.7 was administered to 1 clinically normal dog, 7 dogs with osteogenic sarcoma, 1 dog with undifferentiated sarcoma, and 2 dogs with mammary tumor. Scintigraphy revealed concentration of radioactivity in 8 of 10 tumor sites within 24 hours after MAB administration. The ratio of 131I in tumor sites to 131I in the surrounding normal tissues, compared with the similar ratio of 99mTc-labeled erythrocytes ranged from 1.1 to 4.3, in tumor vs normal tissue with a mean value of 2, confirming tumor localization of the radiolabeled MAB in excess of that associated with enhanced tumor vascularization.

Use of technetium Tc 99m sestamibi for detection of a parathyroid adenoma in a dog with primary hyperparathyroidism.

Double-phase parathyroid gland scintigraphy, using technetium Tc 99m sestamibi, correctly identified the existence and location of a parathyroid adenoma in a dog with primary hyperparathyroidism. The parathyroid adenoma was removed surgically 2 days after scintigraphy. An area of focal radionuclide uptake persisted in the region corresponding to the left external parathyroid gland in the delayed-phase image. Delayed-phase images from 3 healthy dogs and a dog with hypercalcemia of malignancy caused by lymphoma did not reveal an area of persistent radiotracer uptake. Double-phase parathyroid gland scintigraphy, using 99mTc-sestamibi, is a simple, rapid, noninvasive test, which can be used for detection and localization of parathyroid adenomas in hypercalcemic dogs. It also can help to differentiate these dogs from dogs with hypercalcemia of malignancy.

Management of primary headache in emergency services of Santos and surrounding towns.

OBJECTIVES: Primary headaches are often seen by Clinicians on duty at Emergency Services. We have investigated the treatment of such patients by 43 medical doctors who have been working at Emergency Services in the city of Santos and surrounding towns for many years. RESULTS: We confirmed the high prevalence of primary headaches in Emergency Services. There seem to be diagnosis difficulties concerning differentiating attacks of migraine and tension type headache. We also observed that IV dipirone was the most frequently prescribed treatment for patients with primary headaches in this study. There is no protocol in the literature which recommends IV dipirone for the treatment of migraine attacks or other primary headaches. CONCLUSION: It would be advisable to perform controlled double blind studies in order to verify the advantages of IV dipirone in the treatment of intense attacks primary headaches. We concluded that headache management recycling programs could be of interest for doctors who regularly work at Emergency Services.

Single pulse frequency compounding protocol for superharmonic imaging.

Second harmonic imaging is currently accepted as the standard in commercial echographic systems. A new imaging technique, coined as superharmonic imaging (SHI), combines the third till the fifth harmonics, arising during nonlinear sound propagation. It could further enhance the resolution and quality of echographic images. To meet the bandwidth requirement for SHI a dedicated phased array has been developed: a low frequency subarray, intended for transmission, interleaved with a high frequency subarray, used in reception. As the bandwidth of the elements is limited, the spectral gaps in between the harmonics cause multiple (ghost) reflection artifacts. A dual-pulse frequency compounding method aims at suppressing those artifacts at a price of a reduced frame rate. In this study we explore a possibility of performing frequency compounding within a single transmission. The traditional frequency compounding method suppresses the ripples by consecutively emitting two short Gaussian bursts with a slightly different center frequency. In the newly proposed method, the transmit aperture is divided into two parts: the first half is used to send a pulse at the lower center frequency, while the other half simultaneously transmits at a slightly higher center frequency. The suitability of the protocol for medical imaging applications in terms of the steering capabilities was performed in a simulation study with INCS and the hydrophone measurements. Moreover, an experimental study was carried out to find the optimal parameters for the clinical imaging protocol. The latter was subsequently used to obtain the images of a tissue mimicking phantom containing strongly reflecting wires. Additionally, the images of a human heart in the parasternal projection were acquired. The scanning aperture with the developed protocol amounts to approximately 90°, which is sufficient to capture the cardiac structures in the standard anatomical projections. The theoretically estimated and experimentally measured grating lobe levels are equal to -28.3 dB and -35.9 dB, respectively. A considerable improvement in the axial resolution of the SHI component (0.73 mm) at -6 dB in comparison with the third harmonic (2.23 mm) was observed. A similar comparison in terms of the lateral resolution slightly favored the superharmonic component by 0.2 mm. Additionally, the images of the tissue mimicking phantom exhibited the absence of the multiple reflection artifacts. The in-vivo acquisition allows one to clearly observe the dynamic of the mitral valve leaflets. The new method is equally effective in eliminating the ripple artifacts associated with SHI as the dual-pulse technique, while the full frame rate is maintained.

Emergence of Klebsiella pneumoniae carrying the novel extended-spectrum beta-lactamase gene variants bla(SHV-40), bla(TEM-116) and the class 1 integron-associated bla(GES-7) in Brazil.

A clinical Klebsiella pneumoniae isolate carrying the extended-spectrum beta-lactamase gene variants bla(SHV-40), bla(TEM-116) and bla(GES-7) was recovered. Cefoxitin and ceftazidime activity was most affected by the presence of these genes and an additional resistance to trimethoprim-sulphamethoxazole was observed. The bla(GES-7) gene was found to be inserted into a class 1 integron. These results show the emergence of novel bla(TEM) and bla(SHV) genes in Brazil. Moreover, the presence of class 1 integrons suggests a great potential for dissemination of bla(GES) genes into diverse nosocomial pathogens. Indeed, the bla(GES-7) gene was originally discovered in Enterobacter cloacae in Greece and, to our knowledge, has not been reported elsewhere.

Risk factors for colonisation of newborn infants during an outbreak of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae in an intermediate-risk neonatal unit.

We describe a cross-sectional survey to identify risk factors for colonisation of neonates by extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae. This occurred following exposure to a colonised healthcare worker during an outbreak in an intermediate-risk neonatal unit. In total, 120 neonates admitted consecutively during a three-month period were screened for ESBL-producing K. pneumoniae by rectal swabbing and 27 were identified as colonised. Multivariate analysis showed colonisation to be independently associated with use of antibiotics and absence of breastfeeding. Previous use of antibiotics presented an odds ratio (OR) of 12.3 [95% confidence interval (CI): 3.66-41.2, P<0.001]. The most commonly used antibiotics were penicillin and amikacin. Breastfeeding was associated with reduced risk for colonisation (OR: 0.22; 95% CI: 0.05-0.99; P=0.049). Nine isolates recovered during the first stage of the outbreak and 27 isolates from surveillance cultures were typed thereafter by pulsed-field gel electrophoresis, revealing six different profiles (A-F). Clones A, C, and E were implicated in the first stage of the outbreak, whereas among the 27 strains recovered from surveillance cultures, all six clones were identified. Clone A was also found on the hand of a nursing auxiliary with onychomycosis. We concluded that prior antimicrobial use predisposed to colonisation. The possible role of breastfeeding as a protective factor needs to be further elucidated. Detection of different genotypes of ESBL-producing K. pneumoniae suggests that dissemination of mobile genetic elements bearing the ESBL gene may have been superimposed on the simple dissemination of a clone during the outbreak.

Immunolocalization of hepatic metastases of human colonic cancer by chimeric anti-TAG72 antibody in SCID mice.

TAG72 is a well-characterized, human tumor-associated antigen present in > 85% of human colonic cancers. In this study, we established an animal model of hepatic metastases of human colonic carcinoma. The high-mucin variant cell line, designated HM7, was derived from the human colonic carcinoma cell line LS174T. Following intrasplenic injection, HM7 was able to induce much greater hepatic metastases in SCID mice compared to its parental cell line LS174T. Numerous hepatic metastases were evident 18 days subsequent to the intrasplenic injection of tumor cells. Using the chimeric anti-TAG72 antibody ccM4, immunohistochemistry demonstrated strong expressions of the TAG72 antigen in these metastases. Our biodistribution and imaging data also showed that the radiolabelled ccM4 antibody was able to localize hepatic metastases in the SCID mice. Based upon these findings, w anticipate that the herein described SCID mouse model will prove most useful in studying hepatic metastases of human colonic carcinoma by using anti-TAG72 therapeutic immunoreagents.