Binary Targeting of siRNA to Hematologic Cancer Cells In Vivo using Layer-by-Layer Nanoparticles.

Using siRNA therapeutics to treat hematologic malignancies has been unsuccessful because blood cancer cells exhibit remarkable resistance to standard transfection methods. Herein we report the successful delivery of siRNA therapeutics with a dual-targeted, layer-by-layer nanoparticle (LbL-NP). The LbL-NP protects siRNA from nucleases in the bloodstream by embedding it within polyelectrolyte layers that coat a polymeric core. The outermost layer consists of hyaluronic acid (a CD44-ligand) covalently conjugated to CD20 antibodies. The CD20/CD44 dual-targeting outer layer provides precise binding to blood cancer cells, followed by receptor-mediated endocytosis of the LbL-NP. We use this siRNA delivery platform to silence B-cell lymphoma 2 (BCL-2), a pro-survival protein, in vitro and in vivo. The dual-targeting approach significantly enhanced internalization of BCL-2 siRNA in lymphoma and leukemia cells, which led to significant downregulation of BCL-2 expression. Systemic administration of the dual-targeted, siRNA-loaded nanoparticle induced apoptosis and hampered proliferation of blood cancer cells both in cell culture and in orthotopic non-Hodgkin's lymphoma animal models. These results provide the basis for approaches to targeting blood-borne cancers and other diseases, and suggest that LbL nanoassemblies are a promising approach for delivering therapeutic siRNA to hematopoetic cell types that are known to evade transfection by other means.

Isolated cervical Cutibacterium acnes osteomyelitis in a patient with no primary source of infection - A case report and review of the literature.

Background: Cervical vertebral osteomyelitis (CVO) is a rare pathology that leads to progressive osseous degradation and eventual loss of bone putting the patient at risk of devastating neurological injury in the event of bony collapse or instability. Cutibacterium acnes formerly called Propionibacterium acnes is rare, but within the last two decades has been an increasingly reported cause of osteomyelitis. The majority of C. acnes vertebral osteomyelitis cases have been reported in patients with a history of prior invasive procedures where direct contamination at the time of procedure was suspected as the underlying etiology. Case Description: We report a unique case of an otherwise healthy 39-year-old male with no prior history of invasive procedures who presented with CVO secondary to C. acnes. He underwent surgical debridement and fusion in conjunction with antibiotic treatment. The patient recovered well and a 2-year follow-up with serial imaging showed no evidence of disease recurrence. Conclusion: C. acnes is an under-recognized and under-reported etiology of spine infections. Clinicians should be aware of the pathological potential and atypical presentation of C. acnes vertebral osteomyelitis.

Tandem Suzuki Coupling/Intramolecular Oxetane Ring Opening to Form Polycyclic Ring Systems.

A tandem one-pot reaction featuring a cross-coupling followed by an intramolecular oxetane ring opening by mild nucleophiles is reported. The overall transformation comprises a carbon-carbon bond formation along with a carbon-heteroatom bond construction providing diverse multicyclic ring systems with a pendant hydroxymethyl handle for further elaboration. This approach constitutes a convergent method for rapid access to various scaffolds. Furthermore, a comparison of computed low-energy conformers is presented to rationalize instances in which cyclization was not observed.

Mild Intramolecular Ring Opening of Oxetanes.

Oxetanes have been increasingly used as stable motifs in medicinal chemistry as well as versatile synthetic intermediates. Herein, an intramolecular ring opening of oxetane carboxamides with mild nucleophiles, such as nitrogen heterocycles, is presented. The reaction proceeds under metal-free basic conditions which is highly unusual in ring opening reactions of oxetanes. Amide formation and oxetane ring opening/cyclization in a one-pot approach affords high levels of molecular complexity in a single step from simple, readily available substrates.

A small-molecule fusion inhibitor of influenza virus is orally active in mice.

Recent characterization of broadly neutralizing antibodies (bnAbs) against influenza virus identified the conserved hemagglutinin (HA) stem as a target for development of universal vaccines and therapeutics. Although several stem bnAbs are being evaluated in clinical trials, antibodies are generally unsuited for oral delivery. Guided by structural knowledge of the interactions and mechanism of anti-stem bnAb CR6261, we selected and optimized small molecules that mimic the bnAb functionality. Our lead compound neutralizes influenza A group 1 viruses by inhibiting HA-mediated fusion in vitro, protects mice against lethal and sublethal influenza challenge after oral administration, and effectively neutralizes virus infection in reconstituted three-dimensional cell culture of fully differentiated human bronchial epithelial cells. Cocrystal structures with H1 and H5 HAs reveal that the lead compound recapitulates the bnAb hotspot interactions.

Outbreak of multidrug-resistant Serratia marcescens infection in a neonatal intensive care unit.

BACKGROUND: Serratia marcescens causes healthcare-associated infections and significant morbidity and mortality in neonatal intensive care units (NICUs). We report the investigation and control of an outbreak of multidrug-resistant (MDR) S. marcescens infection at an NICU. METHODS: An outbreak investigation and a case-control study were undertaken at a 36-bed NICU in a tertiary care hospital in Baltimore, Maryland, for the period from October 2004 through February 2005. The outbreak investigation included case identification, review of medical records, environmental cultures, patient surveillance cultures, personnel hand cultures, and pulsed-field gel electrophoresis (PFGE). The case-control study included case identification and review of medical records. Infection control measures were implemented. Eighteen NICU neonates had cultures that grew MDR S. marcescens during the study period. The case-control study included 16 patients with the outbreak strain or an unidentified strain of MDR S. marcescens and 32 control patients not infected and/or colonized with MDR S. marcescens, treated in the NICU for at least 48 hours during the study period. RESULTS: PFGE analysis identified a single strain of MDR S. marcescens that infected or colonized 15 patients. Two patients had unique strains, and 1 patient's isolate could not be subtyped. An unrelated MDR S. marcescens isolate was recovered from a sink drain. Exposure to inhalational therapy was an independent risk factor for MDR S. marcescens acquisition after adjusting for birth weight. Extensive investigation failed to reveal a point source for the outbreak. CONCLUSION: A single epidemic strain of MDR S. marcescens spread rapidly and threatened to become endemic in this NICU. Transient carriage on the hands of healthcare personnel or on respiratory care equipment was the likely mode of transmission. Cohorting patients and staff, at the cost of bed closures and additional personnel, interrupted transmission and halted the outbreak.

Phenylpiperidine-benzoxazinones as urotensin-II receptor antagonists: synthesis, SAR, and in vivo assessment.

Various 4-phenylpiperidine-benzoxazin-3-ones were synthesized and biologically evaluated as urotensin-II (U-II) receptor antagonists. Compound 12i was identified from in vitro evaluation as a low nanomolar antagonist against both rat and human U-II receptors. This compound showed in vivo efficacy in reversing the ear-flush response induced by U-II in rats.

Systemic actinomycosis mimicking pelvic malignancy with pulmonary metastases.

A 40-year-old woman presented with profound weight loss, anemia and multiple pulmonary nodules. Further investigation revealed a hepatic abscess and a pelvic mass associated with an intrauterine device. Cultures of the hepatic abscess and from peripheral blood demonstrated Actinomyces species. The patient underwent an exploratory laparotomy and hysterectomy. Pathological examination showed characteristic sulphur granules of actinomycosis. Long-term therapy with high-dose intravenous penicillin resulted in clinical and radiographic improvement.

Granulomatous bronchiolitis with necrobiotic pulmonary nodules in Crohn's disease.

A 37-year-old man with extensive Crohn's disease of the stomach, small and large intestine for almost a decade developed respiratory symptoms and radiological findings suggestive of pneumonia that failed to resolve with antibiotic treatment. Computed tomography scanning of his lungs showed extensive changes with cavitated parenchymal nodules. Histological evaluation of an open lung biopsy showed granulomatous bronchiolitis and pulmonary necrobiosis. Treatment with steroids and immunosuppression resulted in complete resolution of his clinical symptoms of pneumonia and abnormal computed tomography imaging changes. Granulomatous bronchiolitis and necrobiotic nodules may be a manifestation of Crohn's disease in the absence of microbial agents, including mycobacteria or fungal agents. While a multiplicity of complex pulmonary changes may occur in Crohn's disease, their clinical recognition and precise pathological definition may be particularly important if treatment with a biological agent, such as infliximab, is being considered.

Frontiers in pulmonary hypertension in infants and children with bronchopulmonary dysplasia.

Pulmonary hypertension (PH) is an increasingly recognized complication of premature birth and bronchopulmonary dysplasia (BPD), and is associated with increased morbidity and mortality. Extreme phenotypic variability exists among preterm infants of similar gestational ages, making it difficult to predict which infants are at increased risk for developing PH. Intrauterine growth retardation or drug exposures, postnatal therapy with prolonged positive pressure ventilation, cardiovascular shunts, poor postnatal lung and somatic growth, and genetic or epigenetic factors may all contribute to the development of PH in preterm infants with BPD. In addition to the variability of severity of PH, there is also qualitative variability seen in PH, such as the variable responses to vasoactive medications. To reduce the morbidity and mortality associated with PH, a multi-pronged approach is needed. First, improved screening for and increased recognition of PH may allow for earlier treatment and better clinical outcomes. Second, identification of both prenatal and postnatal risk factors for the development of PH may allow targeting of therapy and resources for those at highest risk. Third, understanding the pathophysiology of the preterm pulmonary vascular bed may help improve outcomes through recognizing pathways that are dysregulated in PH, identifying novel biomarkers, and testing novel treatments. Finally, the recognition of conditions and exposures that may exacerbate or lead to recurrent PH is needed to help with developing treatment guidelines and preventative strategies that can be used to reduce the burden of disease.

Nonpeptide urotensin-II receptor antagonists: a new ligand class based on piperazino-phthalimide and piperazino-isoindolinone subunits.

We have discovered two related chemical series of nonpeptide urotensin-II (U-II) receptor antagonists based on piperazino-phthalimide (5 and 6) and piperazino-isoindolinone (7) scaffolds. These structure types are distinctive from those of U-II receptor antagonist series reported in the literature. Antagonist 7a exhibited single-digit nanomolar potency in rat and human cell-based functional assays, as well as strong binding to the human U-II receptor. In advanced pharmacological testing, 7a blocked the effects of U-II in vitro in a rat aortic ring assay and in vivo in a rat ear-flush model. A discussion of U-II receptor antagonist pharmacophores is presented, and a specifically defined model is suggested from tricycle 13, which has a high degree of conformational constraint.

Bradycardia and hypotension associated with fomepizole infusion during hemodialysis.

UNLABELLED: We report a case of hypotension and bradycardia associated with intravenous fomepizole infusion. CASE REPORT: A 59-year-old man presented to hospital 10 hours after ethylene glycol ingestion with ataxia, slurred speech, metabolic acidosis, heart rate 70/min, blood pressure 160/100 mmHg. Treatment with hemodialysis and fomepizole began 7.5 hours after admission. Severe bradycardia (29/min) and hypotension (69 mmHg systolic) occurred immediately following a 30 minute intravenous infusion of the first (19 mg/kg) fomepizole dose, but rapidly corrected with 1 mg atropine. Transient bradycardia (48/min) and hypotension (89/57 mmHg) recurred immediately after the second (10 mg/kg) fomepizole dose, also given during dialysis. DISCUSSION: Hemodialysis may cause a drop in blood pressure and heart rate; however, the close temporal relationship with fomepizole infusions, dose-related symptom intensity and recurrence with rechallenge suggest a causal relationship with fomepizole. Hemodialysis, acidosis and high initial fomepizole dose may have enhanced patient susceptibility, as a post-dialysis fomepizole dose was well tolerated. CONCLUSION: Fomepizole may precipitate bradycardia and/or hypotension during hemodialysis. Monitor vital signs closely during and immediately after infusion.

Selection of a 2-azabicyclo[2.2.2]octane-based alpha4beta1 integrin antagonist as an inhaled anti-asthmatic agent.

The alpha4beta1 integrin, expressed on eosinophils and neutrophils, induces inflammation in the lung by facilitating cellular infiltration and activation. From a number of potent alpha4beta1 antagonists that we evaluated for safety and efficacy, 1 was selected as a lead candidate for anti-asthma therapy by the inhalation route. We devised an optimized stereoselective synthesis to facilitate the preparation of a sufficiently large quantity of 1 for assessment in vivo. Administration of 1 to allergen-sensitive sheep by inhalation blocked the late-phase response of asthma and abolished airway hyper-responsiveness at 24h following the antigen challenge. Additionally, the recruitment of inflammatory cells into the lungs was inhibited. Administration of 1 to ovalbumin-sensitized guinea pigs intraperitoneally blocked airway resistance and inhibited the recruitment of inflammatory cells.