Characterization of adaptation of an avian influenza A (H5N2) virus to a mammalian host

We have used the mouse model to monitor the acquisition of virulence of a non-pathogenic influenza A virus upon adaptation to a new mammalian host. An avian strain, A/Mallard duck/Pennsylvania/10218/84 (H5N2) (Mld/PA/84) was adapted to mice by 23 serial lung-to-lung passages until a highly virulent mouse-adapted (MA) variant (Mld/PA/84-MA) emerged. This MA variant was characterized and compared to the parental strain as well as some of its intermediate passage variants. MA variant caused bronchopneumonia in mice with a high mortality rate (the virulence of Mld/PA/84-MA measured as log (EID50/LD50) was 1.75), while the parental, avirulent strain Mld/PA/84 did not cause illness and mortality in mice (log (EID50/LD50) was 7.25). Hemagglutination-inhibition (HAI) test with a set of hemagglutinin- (HA) specific monoclonal antibodies (MAbs) revealed antigenic differences between the parental strain and MA variant. Mld/PA/84-MA reacted with HA-specific MAbs in higher titers than the parental strain. The HA genes of the parental strain Mld/PA/84, the 1st, 3rd, 8th, and 15th intermediate passage variants, and Mld/PA/84-MA were sequenced. Three amino acid changes at positions 203, 273 and 320 were determined in the HA of MA variant. The first of them, Leu-->Pro (320), appeared in the HA stem region at the 8th passage. Two other in the HA1 globular region (Ser-->Phe (203) and Glu-->Gly (273)) appeared at the 15th passage. All of these substitutions were associated with the increase of viral infectivity for mouse lungs and changes in the HA antigenicity. The potential role of these changes in HA with respect to the process of viral interspecies transmission and acquisition of virulence for new host is discussed.     

Novel PRKD gene rearrangements and variant fusions in cribriform adenocarcinoma of salivary gland origin

Polymorphous low‐grade adenocarcinoma (PLGA) and cribriform adenocarcinoma of minor salivary gland (CAMSG) are low‐grade carcinomas arising most often in oral cavity and oropharynx, respectively. Controversy exists as to whether these tumors represent separate entities or variants of one spectrum, as they appear to have significant overlap, but also clinicopathologic differences. As many salivary carcinomas harbor recurrent translocations, paired‐end RNA sequencing and FusionSeq data analysis was applied for novel fusion discovery on two CAMSGs and two PLGAs. Validated rearrangements were then screened by fluorescence in situ hybridization (FISH) in 60 cases. Histologic classification was performed without knowledge of fusion status and included: 21 CAMSG, 18 classic PLGA, and 21 with “mixed/indeterminate” features. The RNAseq of 2 CAMSGs showed ARID1A‐PRKD1 and DDX3X‐PRKD1 fusions, respectively, while no fusion candidates were identified in two PLGAs. FISH for PRKD1 rearrangements identified 11 additional cases (22%), two more showing ARID1A‐PRKD1 fusions. As PRKD2 and PRKD3 share similar functions with PRKD1 in the diacylglycerol and protein kinase C signal transduction pathway, we expanded the investigation for these genes by FISH. Six additional cases each showed PRKD2 and PRKD3 rearrangements. Of the 26 (43%) fusion‐positive tumors, there were 16 (80%) CAMSGs and 9 (45%) indeterminate cases. A PRKD2 rearrangement was detected in one PLGA (6%). We describe novel and recurrent gene rearrangements in PRKD1–3 primarily in CAMSG, suggesting a possible pathogenetic dichotomy from “classic” PLGA. However, the presence of similar genetic findings in half of the indeterminate cases and a single PLGA suggests a possible shared pathogenesis for these tumor types. © 2014 Wiley Periodicals, Inc.     

Electro-encephalogram disturbances in different sleep-wake states following exposure to high environmental heat

In this study, cerebral electrical activity or electro-encephalogram (EEG) was studied following exposure to high environmental heat, in three different age groups of freely moving rats. Each age group was subdivided into three groups: the acute heat stress group, subjected to a single exposure of 4h at 38°C in the biological oxygen demand incubator; the chronic heat stress group, exposed for 21 days, for 1 h each day, at 38°C in the incubator; and the handling control group. The polygraphic sleep-wake recordings involved simultaneous recordings of cortical EEG, electrooculogram (EOG), and electromyogram (EMG), on paper and in digital form on computer hard disk, just after the heat exposure for the acute stressed rats and on the 22nd day for the chronic stressed rats. The power spectrum was calculated for 2s epochs of the EEG signals. Quantitative analyses of EEG (qEEG) showed that, in all three age groups, changes in higher-frequency components (β₂) were significant in all sleep-wake states following both acute and chronic heat stress conditions. The power of β₂ activity in all three age groups after acute heat exposure was significantly decreased during slow wave sleep (SWS) (p<0.05) and rapid eye movement sleep (p<0.05), whereas the reverse was observed in the awake state (p<0.05). Following chronic heat exposure, β₂ activity was found to increase in all three sleep-wake stages in all groups of rats (p<0.01 for SWS in the weaning group and p<0.05 for other data). Thus the study demonstrated that the cortical EEG is sensitive to environmental heat, and alterations in EEG frequencies in different states of mental consciousness due to high heat can be differentiated efficiently by EEG power spectrum analysis.     

Prevalence and characteristics of type 2 diabetes mellitus in 9-18 year-old children with diabetic ketoacidosis

OBJECTIVE: To estimate the prevalence of type 2 diabetes mellitus (DM2) in 9-18 year-old children with diabetic ketoacidosis (DKA) and to describe the presenting biochemical characteristics and response to standardized DKA treatment. METHODS: Data were collected from a consecutive sample of 9-18 year-old children presenting with DKA. DKA was defined as hyperglycemia and ketosis with an initial pH <7.3, or bicarbonate <15 mmol/l. Patients were classified as having DM2 if they had negative autoantibody status and normal or elevated fasting C-peptide. RESULTS: The prevalence of DM2 in patients with DKA was 13.0% (6.1-23.3%). There was no significant difference in the presenting pH (7.14 vs 7.15), but blood glucose was higher (735 vs 587 mg/dl) in patients with DM2, than in patients with type 1 DM (DM1). The duration of insulin infusion until resolution of acidosis (17.3 vs 13.2 h) and intensive care unit stay (2.4 vs 1.6 days) were longer in patients with DM2. Seven of the nine patients with DM2 did not require insulin at 1-year follow-up. CONCLUSIONS: Children with DM2 can present with DKA and constitute a significant percentage in the above 9-year age group. The need for insulin must be carefully re-evaluated as DKA resolves in these patients. Adolescents with DM2 and their families need to be educated about DKA.