Small molecule inhibitors of the host cell COX/AREG/EGFR/ERK pathway attenuate cytomegalovirus-induced pathogenesis

As with other herpesviruses, human cytomegalovirus (hCMV) has the ability to establish lifelong persistence and latent infection following primary exposure, salivary glands (SMGs) being the primary site of both. In the immunocompromised patient, hCMV is a common cause of opportunistic infections, and subsequent morbidity and mortality. Elucidating the molecular pathogenesis of CMV-induced disease is critical to the development of more effective and safer drug therapies. In the present study, we used a novel mouse postnatal SMG organ culture model of mCMV-induced dysplasia to investigate a candidate signaling network suggested by our prior studies (COX-2/AREG/EGFR/ERK). The objective was to employ small molecule inhibitors to target several key steps in the autocrine loop, and in this way ameliorate pathology. Our results indicate that upregulation of ERK phosphorylation is necessary for initial mCMV-induced pathogenesis, and that ErbB receptor family phosphorylation and downstream signaling are highly relevant targets for drug discovery.     

Comparison of Cuffed and Uncuffed Endotracheal Tubes in Young Children during General Anesthesia

Background: Uncuffed endotracheal tubes are routinely used in young children. This study tests a formula for selecting appropriately sized cuffed endotracheal tubes and compares the use of cuffed versus uncuffed endotracheal tubes for patients whose lungs are mechanically ventilated during anesthesia.

Methods: Full-term newborns and children (n = 488) through 8 yr of age who required general anesthesia and tracheal intubation were assigned randomly to receive either a cuffed tube sized by a new formula [size(mm internal diameter) = (age/4) + 3], or an uncuffed tube sized by the modified Cole's formula [size(mm internal diameter) = (age/4) + 4]. The number of intubations required to achieve an appropriately sized tube, the need to use more than 21 [center dot] min sup -1 fresh gas flow, the concentration of nitrous oxide in the operating room, and the incidence of croup were compared.

Results: Cuffed tubes selected by our formula were appropriate for 99% of patients. Uncuffed tubes selected by Cole's formula were appropriate for 77% of patients (P < 0.001). The lungs of patients with cuffed tubes were adequately ventilated with 2 1 [center dot] min sup -1 fresh gas flow, whereas 11% of those with uncuffed tubes needed greater fresh gas flow (P < 0.001). Ambient nitrous oxide concentration exceeded 25 parts per million in 37% of cases with uncuffed tubes and in 0% of cases with cuffed tubes (P < 0.001). Three patients in each group were treated for croup symptoms (1.2% cuffed; 1.3% uncuffed).     

A wrinkled structure of gold film greatly improves the signaling of electrochemical aptamer-based biosensors

Electrochemical aptamer-based (E-AB) sensors provide a great opportunity towards the goal of point-of-care and wearable sensing technologies due to their good sensitivity and selectivity. Nevertheless, the output signals from this sensor class remain low when sensors are interrogated via square-wave voltammetry. This low signaling limits the sensor''s precision for its capability to detect small changes in target concentrations. To circumvent this, we proposed here the use of a readily shrink-induced, wrinkled Au-coating polyolefin film to immobilize a greater number of DNA probes and thus improve the signaling. Specifically, wrinkled gold film exhibits a 5.5-fold increase of surface area in comparison to the unwrinkled ones. Using these substrates we fabricated a set of E-AB sensors of three biological molecules, including kanamycin, doxorubicin and ATP. We achieved up to 10-fold increase in its current and also good accuracies within ±20% error in the target concentration range across 2 orders of magnitude.

A wrinkled gold substrate greatly improves the signaling of electrochemical aptamer-based biosensors, achieving up to 10-fold increase in signals.     

Initial fluid resuscitation for patients with diabetic ketoacidosis: how dry are they?

A prospective consecutive case series of patients aged 5 to 20 years who presented to a pediatric emergency department with diabetic ketoacidosis (DKA) was studied to determine the actual percent loss of body weight during an episode of DKA to determine the degree of dehydration and thereby provide a guide for hydration therapy during such an episode. Patients' weights at the time of presentation, inpatient discharge, and at the first follow-up clinic visit were used to calculate the percent loss of body weight. Data from 33 episodes of DKA showed that the majority of patients with DKA had moderate (4% to 8%) dehydration. Clinical assessment was a poor predictor of severity of dehydration and overestimated the percent dehydration in 67% of patients. Based on these data it is recommend that initial fluid therapy for DKA should assume moderate dehydration with adjustment made according to clinical response.     

Composition of the Inflammatory Infiltrate in Pediatric Penile Lichen Sclerosus Et Atrophicus (Balanitis Xerotica Obliterans): A Prospective, Comparative Immunophenotyping Study

Dermatopathological evaluation of pediatric preputial inflammatory disease rarely allows for specific diagnosis other than pediatric penile lichen sclerosus et atrophicus (balanitis xerotica obliterans, LSA/BXO). A prospective immunopathological study was performed on 20 consecutive, unselected, clinically and histopathologically confirmed LSA/BXO cases to determine the relative presence of T and B lymphocytes. There were seven cases with early stages of disease, eight with florid disease, and five with later stages of disease. Two ritual circumcision specimens and 12 specimens with non-LSA/BXO balanitis, collected during the same period, were used as controls. The infiltrate in LSA/BXO patients was wholly composed of T cells (positive with UCLH-1 antibody) in all cases. B cells (positive with L-26 antibody) were found only focally in small, discreet, easily recognizable (follicular or early follicle-like) aggregates, positioned slightly deeper than the band-like infiltrate of T cells. T cells were inconspicuous in 9 of the 12 control specimens. In the three other controls, T cells were much more obvious and these patients showed clinical features possibly suggestive of LSA/BXO in early, fiediagnosable phases of development. We conclude that limited immunophenotyping may be a useful adjunct to diagnosis in pediatric cases in which onb limited tissue is available 07 the disease may be m e difficult to classify with confidence.     

Modulation of bacterial growth by tumor necrosis factor-alpha in vitro and in vivo

Tumor necrosis factor-alpha (TNF-alpha) plays an important role in innate immunity. Recent in vitro studies have shown that TNF-alpha may also serve as a growth factor for some bacteria. We examined the physiologic relevance of this phenomenon both in vitro and in vivo. Recombinant mouse TNF-alpha increased in vitro proliferation of Escherichia coli but not Pseudomonas aeruginosa in a concentration-dependent manner, and this effect was attenuated by anti-TNF-alpha antibodies. However, in vivo, TNF-alpha gene-deficient (TNF-alpha-/-) mice showed higher mortality than wild-type (TNF-alpha+/+) mice after inoculation of intranasal bacteria. An impaired bacterial clearance in TNF-alpha-/- mice was associated with decreased systemic concentrations of chemokine macrophage inflammatory protein-2, reduced pulmonary neutrophil recruitment, and depressed expression of neutrophil CD11b and CD16/CD32, suggesting that the effect of TNF-alpha on E. coli growth was outweighed by the recruited neutrophils. We also demonstrated that neutropenic TNF-alpha+/+ mice had approximately 100-fold higher E. coli counts in their lungs than TNF-alpha-/- mice, although survival rates in both groups were similar. We conclude that TNF-alpha augments E. coli growth in vitro and in vivo. However, in vivo, this effect becomes only apparent in neutropenic animals. The relevance of these findings for immune compromised patients remains to be investigated.