MR Imaging and Clinical Characteristics of Diffuse Glioneuronal Tumor with Oligodendroglioma-like Features and Nuclear Clusters.

BACKGROUND AND PURPOSE: Diffuse glioneuronal tumor with oligodendroglioma-like features and nuclear clusters (DGONC) is a new, molecularly defined glioneuronal CNS tumor type. The objective of the present study was to describe MR imaging and clinical characteristics of patients with DGONC. MATERIALS AND METHODS: Preoperative MR images of 9 patients with DGONC (median age at diagnosis, 9.9 years; range, 4.2-21.8 years) were reviewed. RESULTS: All tumors were located superficially in the frontal/temporal lobes and sharply delineated, displaying little mass effect. Near the circle of Willis, the tumors encompassed the arteries. All except one demonstrated characteristics of low-to-intermediate aggressiveness with high-to-intermediate T2WI and ADC signals and bone remodeling. Most tumors (n = 7) showed a homogeneous ground-glass aspect on T2-weighted and FLAIR images. On the basis of the original histopathologic diagnosis, 6 patients received postsurgical chemo-/radiotherapy, 2 were irradiated after surgery, and 1 patient underwent tumor resection only. At a median follow-up of 61 months (range, 10-154 months), 6 patients were alive in a first complete remission and 2 with stable disease 10 and 21 months after diagnosis. The only patient with progressive disease was lost to follow-up. Five-year overall and event-free survival was 100% and 86±13%, respectively. CONCLUSIONS: This case series presents radiomorphologic characteristics highly predictive of DGONC that contrast with the typical aspects of the original histopathologic diagnoses. This presentation underlines the definition of DGONC as a separate entity, from a clinical perspective. Complete resection may be favorable for long-term disease control in patients with DGONC. The efficacy of nonsurgical treatment modalities should be evaluated in larger series.

[Risk factors of permanent stoma in rectal cancer patients undergoing transabdominal anterior resection with temporary stoma].

Objective: To investigate the risk factors of turning temporary stoma into permanent stoma in rectal cancer patients undergoing transabdominal anterior resection with temporary stoma. Methods: A case-control study was carried out. Data of rectal cancer patients who underwent transabdominal anterior resection with temporary stoma and completed follow-up in Department of General Surgery of Xiangya Hospital of Central South University from June 2008 to June 2018 were collected and analyzed. In this study, temporary stoma included defunctioning stoma (ostomy was made during operation) and salvage stoma (ostomy was made within one month after operation due to anastomotic leakage or severe complications). Cases of multiple intestinal tumors were excluded. A total of 308 rectal cancer patients were enrolled in the study, including 198 males and 110 females with a median age of 56 (48-65) years. Ninety-four patients received intraperitoneal chemotherapy during operation. Among 308 patients, upper rectal cancer was observed in 64 cases, middle rectal cancer in 89 cases and low rectal cancer in 155 cases. Twenty patients underwent transverse colostomy and 288 underwent ileostomy. Phone call following-up was conducted from August to September 2019 to investigate whether stoma was reversed, causes of reversal failure, and tumor relapsed or not in detail. Permanent stoma was defined as that the stoma was still not reversed by the latest follow-up. The univariate analysis was performed with chi-square test or Fisher's exact test, and variables with P value < 0.10 were included in the non-conditional logistic regression model for multivariate analysis. Results: The median follow-up time was 54.3 (32.4-73.8) months. During follow-up, 8 cases had local recurrence and 37 cases had distant metastasis. Among the 308 patients with temporary ostomy, 247 (80.2%) patients had stomas reversed and the median interval time was 4.5 (3.5-6.1) months. The median interval time in 65 patients with salvage stoma was significantly longer that in 182 patients with defunctioning stoma [5.5 (4.3-7.5) vs. 4.2 (3.4-5.5) months; Z=-4.387, P<0.001]. The temporary ostomy was confirmed to become permanent stoma in 61 patients (19.8%), including 45 cases of defunctioning stoma and 16 cases of salvage stoma. Univariate analysis showed that preoperative anemia, intraperitoneal chemotherapy during operation, middle rectal cancer, transverse colostomy, pathological stage, postoperative local recurrence and distant metastasis were associated with permanent stoma (all P<0.10). Multivariate analysis revealed that the intraperitoneal chemotherapy during operation (OR=1.961, 95% CI: 1.029-3.738, P=0.041), middle rectal cancer (OR=2.401, 95% CI: 1.195-4.826, P=0.014), transverse colostomy (OR=3.433, 95% CI: 1.234-9.553, P=0.018), and distant metastasis (OR=8.282, 95% CI:3.820-17.954, P<0.001) were independent risk factors of permanent stoma. Conclusions: There is high risk of turning temporary stoma into permanent stoma among rectal cancer patients undergoing transabdominal anterior resection who receive intraperitoneal chemotherapy during operation, present as the middle rectal cancer, undergo transverse colostomy or develop distant metastasis. Surgeons need to evaluate and balance the risks and benefits thoroughly, and then inform the patients in order to avoid potential conflicts.

Diffuse glioneuronal tumour with oligodendroglioma-like features and nuclear clusters (DGONC) - a molecularly defined glioneuronal CNS tumour class displaying recurrent monosomy 14.

AIMS: DNA methylation-based central nervous system (CNS) tumour classification has identified numerous molecularly distinct tumour types, and clinically relevant subgroups among known CNS tumour entities that were previously thought to represent homogeneous diseases. Our study aimed at characterizing a novel, molecularly defined variant of glioneuronal CNS tumour. PATIENTS AND METHODS: DNA methylation profiling was performed using the Infinium MethylationEPIC or 450 k BeadChip arrays (Illumina) and analysed using the 'conumee' package in R computing environment. Additional gene panel sequencing was also performed. Tumour samples were collected at the German Cancer Research Centre (DKFZ) and provided by multinational collaborators. Histological sections were also collected and independently reviewed. RESULTS: Genome-wide DNA methylation data from >25 000 CNS tumours were screened for clusters separated from established DNA methylation classes, revealing a novel group comprising 31 tumours, mainly found in paediatric patients. This DNA methylation-defined variant of low-grade CNS tumours with glioneuronal differentiation displays recurrent monosomy 14, nuclear clusters within a morphology that is otherwise reminiscent of oligodendroglioma and other established entities with clear cell histology, and a lack of genetic alterations commonly observed in other (paediatric) glioneuronal entities. CONCLUSIONS: DNA methylation-based tumour classification is an objective method of assessing tumour origins, which may aid in diagnosis, especially for atypical cases. With increasing sample size, methylation analysis allows for the identification of rare, putative new tumour entities, which are currently not recognized by the WHO classification. Our study revealed the existence of a DNA methylation-defined class of low-grade glioneuronal tumours with recurrent monosomy 14, oligodendroglioma-like features and nuclear clusters.

Positive symptoms and white matter microstructure in never-medicated first episode schizophrenia.

BACKGROUND: We investigated cerebral structural connectivity and its relationship to symptoms in never-medicated individuals with first-onset schizophrenia using diffusion tensor imaging (DTI). METHOD: We recruited subjects with first episode DSM-IV schizophrenia who had never been exposed to antipsychotic medication (n=34) and age-matched healthy volunteers (n=32). All subjects received DTI and structural magnetic resonance imaging scans. Patients' symptoms were assessed on the Positive and Negative Syndrome Scale. Voxel-based analysis was performed to investigate brain regions where fractional anisotropy (FA) values significantly correlated with symptom scores. RESULTS: In patients with first-episode schizophrenia, positive symptoms correlated positively with FA scores in white matter associated with the right frontal lobe, left anterior cingulate gyrus, left superior temporal gyrus, right middle temporal gyrus, right middle cingulate gyrus, and left cuneus. Importantly, FA in each of these regions was lower in patients than controls, but patients with more positive symptoms had FA values closer to controls. We found no significant correlations between FA and negative symptoms. CONCLUSIONS: The newly-diagnosed, neuroleptic-naive patients had lower FA scores in the brain compared with controls. There was positive correlation between FA scores and positive symptoms scores in frontotemporal tracts, including left fronto-occipital fasciculus and left inferior longitudinal fasciculus. This implies that white matter dysintegrity is already present in the pre-treatment phase and that FA is likely to decrease after clinical treatment or symptom remission.

Screwworms, Cochliomyia hominivorax, reared for mass release do not carry and spread foot-and-mouth disease virus and classical swine fever virus.

Experiments were done to determine if transporting live screwworms Cochliomyia hominivorax Coquerel (Diptera: Calliphoridae) for developing new strains from countries where foot-and-mouth disease and classical swine fever are endemic, to the mass rearing facilities in Mexico and Panama, may introduce these exotic diseases into these countries. Are screwworms capable of harboring and spreading foot-and-mouth disease virus (FMDV) and classical swine fever virus (CSFV) when they are grown in virus-inoculated larval rearing medium? In one experiment, screwworm larvae were reared in a FMDV-inoculated artificial medium containing either 0.1 % formaldehyde or antibiotics as an antimicrobial agent. In another experiment, larvae were similarly reared in a CSFV-inoculated artificial medium containing 0.1% formaldehyde. In each experiment, samples of larvae and the rearing media were collected daily until pupation occurred. The presence of FMDV was assayed by observing cytopathic effects on cell cultures and a conventional reverse transcription-polymerase chain reaction (RT-PCR); CSFV was assayed using an avidin-biotin complex assay and a conventional RT-PCR. For media containing antibiotics, FMDV was detected in a larval sample collected on day 1 and in media samples on days 1, 2 and 3. No FMDV was detected from larval and media samples collected on all other days. For media containing formaldehyde, FMDV and CSFV were not detectable in larval or media samples collected on all sampling days. These results indicate that FMDV and CSFV cannot survive in rearing medium containing formaldehyde as an antimicrobial agent. Therefore, insects collected in endemic regions and reared using formaldehyde-containing medium for at least one generation at the collection site should be free of FMDV and CSFV and can be transported safely to a strain development/mass rearing facility.

[Coding region cDNA sequence cloning of rat neuroglobin gene, its polymorphism feature and tissue expression profile analysis].

The coding region cDNA sequence of rat neuroglobin (NGB) was obtained by RT-PCR technique using a degeneracy PCR primer pair based on previously reported cDNA sequence of human and mouse NGB gene. Result demonstrated that the coding region cDNA sequence of rat NGB gene is 456 bp in length, which could encode a protein of 151 amino acids. The rat NGB gene is highly homology with mouse (96%) and human (88%) NGB gene. However, several polymorphism sites were also detected in the rat NGB coding region: 113 t/c [L38P], 133 a/g [N45D], 388 a/g[R130G], 417 t/c. The cDNA sequence of rat NGB gene has been registered in GenBank under the accession number AF333245. Moreover, highly expression level of rat NGB in brain, liver, kidney, heart and skeletal muscle was detected by using multiple tissue RT-PCR technique, indicating the functional importance of this novel gene.

Magnetic immunoseparation PCR assay (MIPA) for detection of hepatitis A virus (HAV) in American oyster (Crassostrea virginica).

In order to detect the low numbers of hepatitis A viral (HAV) particles which may potentially be present in food and cause a serious illness, an original procedure which combines immunomagnetic separation and PCR is described. The use of streptavidin magnetic beads coated with biotinylated human anti-HAV IgG allows virus capture and the removal of the RT-PCR inhibitory compounds which usually are present in shellfish extracts. Following immunomagnetic capture, the separated HAV were lysed, the beads discarded, and the supernatant containing the viral RNA subjected to the RT-PCR protocol. Levels of HAV ranging from 10 to 10(5) pfu were successfully detected in artificially contaminated samples of shucked American oyster (Crassostrea virginica).

Enterotoxigenic Escherichia coli detected in foods by PCR and an enzyme-linked oligonucleotide probe.

A polymerase chain reaction (PCR) and an enzyme-linked oligonucleotide probe hybridization assay were developed for the detection of enterotoxigenic Escherichia coli (ETEC) in ground beef, chicken, pork and raw milk. Two synthetic primers, one of which was biotinylated, were used in the PCR to amplify a fragment of the E. coli heat-labile enterotoxin (LT) gene. The identity of the amplified products was confirmed by liquid hybridization using a horseradish peroxidase-linked internal oligonucleotide probe in a 96-well microplate coated with streptavidin. The final quantitation of the PCR products was performed by a colorimetric reaction. Under established conditions (including 1 min at 60 degrees C for primer annealing and extension in PCR cycles), this method detected all 7 LT-producing E. coli pathogenic for humans, but did not detect all 7 LT-positive E. coli of animal origin 3 E. coli strains that do not produce LT, and 9 other bacteria. Under less stringent PCR conditions (55 degrees C for annealing and extension), 2 strains of LT-producing E. coli of porcine origin were detected while the results of other bacterial strains remained unchanged. In pure cultures, the detection limit of the method was 1.4 colony forming units (CFU). Prior to PCR amplification, all food samples inoculated with an LT-producing ETEC, were subjected to enrichment in brain heart infusion broth for 8 h at 37 degrees C. From these cultures, 10 microliters was heated at 95 degrees C for 10 min and directly used in the PCR. An initial inoculum of as few as 1.2 to 12 CFU of the LT-producing ETEC per 25 g (or ml) of food sample gave a positive reaction.

Detection of Escherichia coli O157:H7 by multiplex PCR.

In order to develop a PCR assay for Escherichia coli O157:H7, a portion of the 60-MDa plasmid harbored by enterohemorrhagic E. coli (EHEC) was sequenced and PCR primers were designed. A multiplex PCR method was then designed by employing primers specific for the EHEC eaeA gene, conserved sequences of Shiga-like toxins I (SLT-I) and II (SLT-II), and the 60-MDa plasmid. PCR products of 1,087 bp (eaeA), 227 and/or 224 bp (SLT-I and/or SLT-II), and 166 bp (plasmid) were successfully amplified simultaneously in a single reaction. The multiplex PCR method can be used to specifically identify EHEC of serogroup O157.

Persistence of inoculated hepatitis A virus in mixed human and animal wastes.

The persistence of hepatitis A virus (HAV) was determined both in mixtures of septic tank effluent (STE) with dairy cattle manure slurry (DCMS) and in mixtures of STE with swine manure slurry (SMS). HAV was consistently inactivated more rapidly in the two types of mixed wastes than in STE alone or in the control Dulbecco's phosphate-buffered saline (PBS). At 5 degrees C, the D values (time, in days, for a 90% reduction of virus titer) were 34.6 for the mixed STE and DCMS, 48.5 for the mixed STE and SMS, 58.5 for STE, and 217.4 for the Dulbecco's PBS control. At 22 degrees C, the D values were 23.0, 17.1, 35.1, and 90.1 for the four suspension media, respectively. A comparison of HAV inactivation in mixed wastes subjected to different treatments at the same pH and temperatures showed that the virus inactivation in the mixed wastes was related, at least in part, to microbial activity. In mixed STE and DCMS, the D values at 25 degrees C were 8.3 for raw mixed wastes, 15.1 for autoclaved mixed wastes, and 9.6 for bacterium-free filtrate of raw mixed wastes; D values at 37 degrees C were 6.8, 10.1, and 7.0 for these three suspension media, respectively. In mixed STE and SMS, the D values at 25 degrees C were 8.1 for raw mixed wastes, 14.3 for autoclaved mixed wastes, and 9.1 for bacterium-free filtrate of raw mixed wastes; the D values at 37 degrees C were 6.8, 9.4, and 6.9 for the three suspensions, respectively.

Antiviral effects of bacteria isolated from manure.

The objectives of this study were to determine the role of microbial activity in inactivation of hepatitis A virus (HAV) and to learn how the virus is inactivated. Of 31 bacterial strains isolated from animal manure, 10 efficiently inactivated HAV in fluid thioglycollate medium, with D10 values (time, in days, required for a 90% reduction of virus titer) of ≤ 10 at 30°C. The D10 value of the control suspension without bacteria was 35.1. Most of the 10 strains raised the pH of the medium during growth; comparisons suggested that alkalinity was not a principal antiviral property of these cultures. Cell-free filtrates of nine of these strains caused net 90% inactivation of HAV within 6 days at 37°C; the other did not. The inactivation capacity of four of the nine culture filtrates was significantly reduced by incubation with selected protease inhibitors before the virus was added. These protease inhibitors did not affect the activities of the other five culture filtrates. Fractions prepared by ultrafiltration (nominal molecular weights <1,000) from two of these cultures inactivated HAV suggesting that their mode of action was not enzymatic.

Detection of hepatitis A virus in environmental samples by antigen-capture PCR.

The efficacy of the antigen-capture PCR (AC-PCR) method for the detection of hepatitis A virus (HAV) in environmental samples was demonstrated. HAV was captured from a seeded liquid waste or a shellfish sample with homologous antibody and then heat denatured and subjected to reverse transcription and the PCR, all in the same tube. Subsequently, the AC-PCR products were analyzed by oligonucleotide probe hybridization in solution, agarose gel electrophoresis, and autoradiography. The AC-PCR detected as little as 0.053 PFU of cell culture-adapted HAV strain HM175/18f. The results of cDNA-RNA hybridization indicated that the particle/PFU ratio of this virus strain is approximately 79:1. Therefore, the detection limit of the AC-PCR was estimated to be four virus particles. No amplified products were observed when poliovirus 1, coxsackievirus A9, coxsackievirus B3, echovirus 6, reovirus 1, adenovirus type 40, human rotavirus type 1, and bovine enterovirus type 2 were tested, confirming the specificity of the assay. There were no differences between the nucleotide sequences of AC-PCR products of HAV strain HM175/18f and the sequences of wild-type HAV strain HM175 derived from molecularly cloned cDNA. Of 121 waste and shellfish samples tested by both plaque assays (PA) in cell cultures and the AC-PCR, 81 (67%) were positive and 31 (26%) were negative as determined by both methods, whereas 9 (7%) were positive as determined by the AC-PCR and negative as determined by the PA, and none were positive as determined by the PA and negative as determined by the AC-PCR.

Degradation of Giardia lamblia cysts in mixed human and swine wastes.

This study was conducted to determine the persistence of Giardia lamblia cysts in mixed septic tank effluent and swine manure slurry and to correlate fluorescein diacetate-propidium iodide staining of G. lamblia cysts with their morphology under low-voltage scanning electron microscopy. Under field conditions, G. lamblia cysts were degraded more rapidly in the mixed waste than in the control Dulbecco's phosphate-buffered saline (PBS). For total and viable cysts, the mixed waste had D values (time for a 90% reduction in number of cysts) of 18.3 and 15.5 days, and the Dulbecco's PBS control had D values of 41.6 and 26.8 days. The rates of cyst degradation in septic tank effluent and in Dulbecco's PBS were similar. Increasing the proportion of swine manure slurry in the mixed waste favored degradation of the parasite. These results indicate that the mixed waste treatment was the predominant factor affecting the cyst persistence and that it was swine manure slurry that played the role of degrading the parasite. Visualization of viable and nonviable Giardia cysts with low-voltage scanning electron microscopy revealed an excellent correlation between the viability of the cysts determined by fluorescein diacetate-propidium iodide staining and their electron microscopic morphology.

Inactivation of poliovirus type 1 in mixed human and swine wastes and by bacteria from swine manure.

The persistence of poliovirus type 1 (PO1) in mixed septic tank effluent and swine manure slurry was determined, and the antiviral effects of several bacterial cultures isolated from swine manure slurry were demonstrated. In two field experiments, PO1 was consistently inactivated more rapidly in the mixed waste than in the control Dulbecco's phosphate-buffered saline (D-PBS). D values (time [in days] for a 90% reduction of virus titer) were 18.7 and 29.9 for the mixed waste and 56.5 and 51.8 for the D-PBS control, respectively. The virus inactivation in the mixed waste was temperature dependent. A comparison of PO1 inactivation in raw mixed waste, autoclaved mixed waste, and bacterium-free filtrate of raw mixed waste at the same pH and temperatures provided an initial demonstration that the virus inactivation in the mixed waste is related, at least in part, to microbial activity. At 25 degrees C, the D value was 6.8 for the mixed waste, 11.2 for the autoclaved mixed waste, and 10.5 for the bacterium-free filtrate of raw mixed waste. At 37 degrees C, D values were 1.3, 3.9, and 3.1 for these three suspending media, respectively. Three bacterial isolates which had shown antiviral effects in a screening test each caused virus inactivation in autoclaved mixed waste, in which the effect of other microorganisms was excluded. Inhibition of PO1 inactivation by protease inhibitors suggests that the virus inactivation in the mixed waste was due in part to proteolytic enzymes produced by bacteria in the waste.

Detection of duck plague virus by reverse passive hemagglutination test.

A reverse passive hemagglutination (RPHA) test was developed to detect duck plague virus (DPV). The technique used sheep erythrocytes stabilized with formaldehyde and pyruvaldehyde and coated with immunoglobulin G (IgG) containing anti-DPV antibody prepared from antiserum produced in sheep. Optimum coating of stabilized erythrocytes occurred at 25 C and pH 4.0 with a concentration of IgG of 20-40 micrograms/ml and a 90-min incubation period. The coated cells were stable for 40 days when stored at 4 C or for at least 4 months (the longest period tested) when frozen at -70 C or -196 C. The RPHA test was conducted at 25 C and read after 3 hours. The high specificity of the test is indicated by the absence of cross-reactions with heterologous virus strains, with specimens prepared from normal duck livers, and with normal chicken embryo chorioallantoic fluid, as well as by the inhibition of hemagglutination only with DPV antiserum. The RPHA test detected six strains of DPV in all virus-containing specimens as well as the immunofluorescence (IF) test did; however, conventional plaque assays (PA) failed to detect virus in five specimens that contained three non-plaque-forming strains of DPV. The mean quantity of DPV that could be detected in the RPHA test was 25 plaque-forming units or 65 fluorescent units per ml. Although the RPHA test was less sensitive than either the PA or the IF test, there was a positive correlation in the titers of DPV antigens between all three tests. The RPHA test is a rapid, simple procedure that is sufficiently sensitive for diagnostic detection of DPV in acute infections, especially in tissues of ducks dying of the disease.