Phase I study of liposome-encapsulated c-raf antisense oligodeoxyribonucleotide infusion in combination with radiation therapy in patients with advanced malignancies.

PURPOSE: Raf proteins are key elements of growth-related cellular signaling pathways and are a component of cancer cell resistance to radiation therapy. Antisense oligonucleotides to c-raf-1 permit highly selective inhibition of the gene product and offer a strategy for sensitizing cancer cells to radiation therapy. In this dose escalation study, we evaluated the safety of combined liposomal formulation of raf antisense oligonucleotide (LErafAON) and radiation therapy in patients with advanced malignancies. EXPERIMENTAL DESIGN: Patients with advanced solid tumors were treated with LErafAON in a phase I dose escalation study while receiving palliative radiation therapy. Drug-related and radiation-related toxicities were monitored. Pharmacokinetics and expression of c-raf-1 mRNA and Raf-1 protein were determined in peripheral blood mononuclear cells. RESULTS: Seventeen patients with palliative indications for radiation therapy were entered into this study. Thirteen patients received daily infusions of LErafAON and four received twice-weekly infusions. Radiation therapy was delivered in daily 300-cGy fractions over 2 weeks. Patients tolerated radiation, and no unexpected radiation-related side effects were observed. Drug-related reactions (grade > or =2), such as back pain, chills, dyspnea, fatigue, fever, flushing, and hypertension, were observed in most patients and were managed by premedication with corticosteroids and antihistamines. Serious adverse events occurred in five patients, including acute infusion-related symptoms, abnormal liver function tests, hypoxia, dehydration, diarrhea, esophagitis, fever, hypokalemia, pharyngitis, and tachypnea. Twelve of 17 patients were evaluable for tumor response at completion of treatment; four showed partial response, four showed stable disease, and four experienced progressive disease. The intact rafAON was detected in plasma for 30 minutes to several hours. Six patients with partial response or stable disease were evaluable for c-raf-1 mRNA and/or Raf-1 protein expression. Inhibition of c-raf-1 mRNA was observed in three of five patients. Raf-1 protein was inhibited in four of five patients. CONCLUSION: This is the first report of the combined modality treatment using antisense oligonucleotides with radiation therapy in patients with advanced cancer. A dose of 2.0 mg/kg of LErafAON administered twice weekly is tolerated with premedication and does not enhance radiation toxicity in patients. The observation of dose-dependent, infusion-related reactions has led to further modification of the liposomal composition for use in future clinical trials.     

Storage of heat-fixed unstained sputum AFB smears for panel testing in a tuberculosis unit in South India.

To evaluate the suitability for panel testing of heat-fixed unstained sputum AFB smears stored for up to 10 months, panels of slides were prepared at the national laboratory and stored under ambient conditions. Every month, three slides were utilised for panel testing in each of 12 microscopy centres; 70 smears were checked in a blinded fashion after 10 months. Reading errors occurred in 15/360 slides used in panel testing and in 4/70 slides used in blinded checking. The quality and grading of heat-fixed unstained smears were unaffected for up to 10 months and were found suitable for panel testing.     

Naturally Inspired Pyrimidines Analogues for Alzheimer’s Disease

Alzheimer’s disease (AD) is a multifarious and developing neurodegenerative disorder. The treatment of AD is still a challenge and availability of drug therapy on the basis of symptoms is not up to the mark. In the context of existence, which is getting worse for the human brain, it is necessary to take care of all critical measures. The disease is caused due to multidirectional pathology of the body, which demands the multi-target-directed ligand (MTDL) approach. This gives hope for new drugs for AD, summarized here in with the pyrimidine based natural product inspired molecule as a lead. The review is sufficient in providing a list of chemical ingredients of the plant to cure AD and screen them against various potential targets of AD. The synthesis of a highly functionalized scaffold in one step in a single pot without isolating the intermediate is a challenging task. In few examples, we have highlighted the importance of this kind of reaction, generally known as multi-component reaction. Multi-component is a widely accepted technique by the drug discovery people due to its high atom economy. It reduces multi-step process to a one-step process, therefore the compounds library can be made in minimum time and cost. This review has highlighted the importance of multicomponent reactions by giving the example of active scaffolds of pyrimidine/fused pyrimidines. This would bring importance to the fast as well as smart synthesis of bio-relevant molecules.     

Financial burden of heart failure in a developing country: cost analysis from Manipal Heart Failure Registry,India

Aim

To estimate the comprehensive healthcare costs of heart failure (HF) and determine the utilization of healthcare resources (HRU) for 2 years following index hospitalization.

Subjects and methods

The Manipal Heart Failure Registry (MHFR) is a prospective registry analyzing the financial burden and HRU in 610 patients with HF. Costs incurred by patients during 2 years following index hospitalization were estimated, and their association with socio-demographic and clinical factors were calculated. After 54 (8.8%) in-hospital mortalities, 556 patients were followed up for 2 years.

Results

The mean age of the study cohort was 65.08?±?13.6 years, and 245 (40.2%) were females. Based on the ejection fraction (EF), 506 (82.9%) patients were diagnosed as having HF with reduced EF. Average hospital stay during index admission was 5.3?±?2.9 days. Total expenditure during index hospitalization was INR 36.3 million and during 2-year follow-up was INR 45.2 million. Average total expenditure per patient was INR 133,663. The average out-of-pocket expense was INR 82,766 and average health insurance coverage was INR 50,896. Difference in expenditure was significant (P?<?0.05) between specific groups, i.e., etiologies, genders, HF phenotypes, age groups, and healthcare insurance types.

Conclusion

Healthcare expenditure of patients with HF in India is much lower than for the western counterparts. Higher utilization of healthcare resources in HF patients with ischemic etiology, non-compliant to medications, and elderly (age?>?60 years) was associated with increased expenses. Interventional procedures and implantations account for the bulk of the expenses in ischemic HF patients.

Trial registration number

Clinical Trial Registry of India: CTRI/2017/11/010395; National Institute of Health (NIH) clinical trial no.: NCT03157219.

     

Intra‐arterial Effects of Cisplatin on Microvascular Anastomoses in the Rat Model

Objective To evaluate the patency of microvascular anastomoses in arteries exposed to intra‐arterial cisplatin. Study Design Animal model. Methods The common iliac artery of 15 rats was injected with 150 mg/m² cisplatin. Five rats were injected with the same volume of saline serving as physiological controls. The ipsilateral femoral artery was transected and anastomosed using microsurgical technique within 3 to 5 days. A Doppler probe was used before and after the anastomosis to assess blood flow. The vessel was re‐examined on postoperative day 5. Pulsatile blood flow and the presence or absence of a Doppler signal was recorded at this time. Vessels were harvested to include the anastomosis site and fixed for histological evaluation. The contralateral femoral artery was also harvested for comparison. Results All femoral artery anastomoses in the experimental and control arm had good, pulsatile blood flow by microscopic evaluation. No thrombosed vessels were visualized, and Doppler signals remained strong at all vessel anastomoses. Histological analysis of the vessels revealed a trend toward increased inflammatory infiltrate in the walls of the vessels treated with cisplatin. We did not appreciate a functional decrease in lumen size. Conclusions Selective catheterization intra‐arterial cisplatin chemotherapy does not affect the patency of vessels following a microvascular anastomosis in the rat model. The trend toward increased inflammatory response in the vessel walls may suggest the need for closer monitoring in patients treated with intra‐arterial chemotherapy.     

Enhanced Cellular Uptake of Oligonucleotides by EGF Receptor-Mediated Endocytosis in A549 Cells

Purpose. The goal of this study was to investigate the feasibility of utilizing epidermal growth factor (EGF) receptor-mediated endocytosis to enhance cellular uptake and targeting of oligonucleotides in epithelial cancer cells. To overcome the problem of endosomal entrappment associated with receptor-mediated delivery, we also examined the effects of two fusogenic peptides, polymyxin B and influenza HA2 peptide, for their capability to promote cytoplasmic delivery of oligonucleotides. Methods. A molecular conjugate consisting of EGF and poly-L-lysine (PL) was synthesized and complexed with 5′ fluorescently-labeled oligonucleotide. Cellular uptake of the complex in presence or absence of the fusogenic peptides was monitored fluorometrically. Microscopic studies were performed to visualize the intracellular distribution of the oligonucleotide. Results. Cells treated with the complex exhibited intracellular fluorescence intensity significantly enhanced over free oligonucleotide-treated controls. The uptake of the complex was shown to occur via the EGF receptor-mediated pathway. Fluorescence microscopic studies revealed cellular internalization of the complex, however, the complex appeared to be accumulated in endocytic vesicles. Exposure of the cells to complex in presence of HA2 peptide and polymyxin B resulted in a more diffused intracellular fluorescence pattern and a corresponding increase in fluorescence intensity. These results are consistent with the known fusion and destabilizing activities of the peptides. Conclusions. Since EGF receptors are overexpressed in many cancer cell types, the EGF-PL conjugate may potentially be used as an effective and selective delivery system to enhance uptake of oligonucleotides into cancer cells.     

Diagnosis and Management of Rhabdomyosarcoma in Children and Adolescents: ICMR Consensus Document

Rhabdomyosarcoma (RMS) is a highly malignant tumor which is thought to originate from the pluripotent mesenchyme. It is the most common soft-tissue sarcoma of childhood. This review article summarizes the recent and older published literature and gives an overview of management of RMS in children. RMS can arise in a wide variety of primary sites, some of which are associated with specific patterns of local invasion, regional lymph nodal spread, therapeutic response and long term outcome, hence requiring physicians to be familiar with site-specific staging and treatment details. Most common primary sites include the head and neck region, genitourinary tract, and extremities. Prognosis for children and adolescents with RMS has recently improved substantially, especially for patients with local or locally extensive disease because of the development of multi-modal therapy incorporating surgery, dose-intensive combination chemotherapy, and radiation therapy. Despite aggressive approaches the outcome for patients who present with metastatic disease remains unsatisfactory. Clinical trials are ongoing to reduce toxicity and improve outcomes of such patients; newer agents in combination are being investigated.     

Growth failure in hereditary spherocytosis and the effect of splenectomy

Objectives

To analyze growth-failure and improvement, if any, following splenectomy in children with hereditary spherocytosis.

Methods

Data collection from case-records (n=82) over 27-years (1985-2011).

Results

Prevalence of stunting was 26%; 32% were underweight. Stunted children were older in age (P=0.006) and presented late (P=0.003). Splenectomy (n=26) improved anemia (P<0.001). However, height-for-age did not improve at 1-year, or 4.5-years (median) following splenectomy (P=1.0). Number of underweight children did not reduce at 1-(P=0.21), or 4.5-years (P=0.21) following surgery.

Conclusion

Growth-failure is frequent in children with hereditary spherocytosis in India. Splenectomy corrected the anemia but failed to improve the growth.