Solution Oligonucleotide APIs: Regulatory Considerations.

Manufacture of oligonucleotide active pharmaceutical ingredients (APIs) typically consists of solid-phase synthesis, deprotection and cleavage, purification and filtration, and isolation from aqueous solutions through lyophilization. In the first step of drug product manufacture, the API is dissolved in water again and excipients are added. While isolation of oligonucleotide APIs can be meaningful in many cases, there may be cases where keeping the API in solution provides benefit, and multiple technical aspects must be taken into account and balanced when determining the appropriate API form. A significant factor is whether an API in solution will contain additional components. While APIs in solution containing additional components (so-called formulated APIs) are well established for biological products, there are regulatory guidelines in place that represent hurdles for industry to using a formulated API approach for oligonucleotide drugs. The present communication outlines conditions where a formulated API approach can be chosen in compliance with existing guidelines. Relevant aspects pertaining to risk management, GMP standards, facility design, control strategies, and regulatory submission content are discussed. In addition, the authors propose that existing guidelines be modernized to enable the use of a formulated API approach for additional reasons than the ones described in the existing regulatory framework. The manuscript aims to promote a dialog with regulators in this field.

An economical and efficient method for postmortem DNA sampling in mass fatalities.

In mass fatality events, the need to identify large numbers of deceased persons using DNA can be a significant drain on already overburdened forensic practitioners, both in the field setting and the laboratory. The laboratory may be required to extract DNA from a variety of postmortem sample types, family or direct reference samples related to the missing, and perform matching of these results in a short period of time. While most forensic institutions are well equipped to handle both family and direct reference samples, postmortem samples such as bone or heterogeneous tissue samples can be difficult for labs to analyze. We have devised an easily deployable, efficient, and inexpensive method for collecting postmortem DNA samples on commercially available DNA preservation cards ("FTA®" cards). FTA® cards are already widely used in forensic labs and are convenient for shipping due to their small volume and stability at room temperature. We evaluated the suitability of a protocol involving swabbing of incisions made on cadavers and sample deposition onto FTA® cards over various postmortem intervals and under different environmental conditions. Each trial took place during a different point in the calendar year to evaluate the effects of seasonal weather patterns and temperature on decomposition, DNA yield, and rates of DNA degradation. To further account for the effects of seasonality (temperature and humidity), the progression of body decomposition was recorded following the Total Body Score (TBS) method [1]. DNA degradation was assessed either through STR amplification of 1.2 mm FTA punches or DNA extraction from 3.0 mm punches followed by real-time PCR quantification and STR amplification and genotyping. No consistent relationship was observed between postmortem interval and DNA degradation. Instead, the TBS score, which captures the stage of body decomposition, was shown to correlate well with DNA quantity. A TBS of 15 and below consistently yielded strong partial or full profiles (20 STR loci and Amelogenin using the PowerPlex 21 System) from all individuals from either 1.2 mm or 3.0 mm punches. Transfer of sample swabs to FTA cards is shown to be a simple and effective method for both field and laboratory operations over a range of conditions that can be evaluated by field forensic practitioners based on a body decomposition score. The approach could be beneficially integrated into mass fatality response plans.

Non-sting barrier cream in radiotherapy-induced skin reactions.

A pilot evaluation was undertaken in 13 patients with head and neck cancer exploring the use of a non-sting barrier film (Sorbaderm(®,) Aspen Medical Europe Ltd). The Society of Radiographers issued guidance in 2013 warning their members that the use of Aqueous cream for moisturising the skin during radiotherapy was potentially harmful. Patients were monitored over a period of 6 weeks and the aim was to explore whether applying non-sting barrier cream provided a protective barrier that did not impair treatment, slowed or prevented skin damage, was easy and simple to apply for patients and carers, improved quality of life for patients during radiotherapy or aided healing. There appeared to be a delay in skin breakdown in this evaluation from week 3 to week 4 and then only mild pain was recorded and with a maximum Radiation Therapy Oncology Group scale of 2.5 in the patient that had combined chemotherapy and radiotherapy. The patients' overall assessment demonstrated that the use of non-sting barrier cream provided symptom relief in both dry, tightening and itching of the skin associated with radiotherapy. All except one patient found the cream easy to apply. The head and neck nursing team rated the product as 'good' to 'very good'.

Polymeric membrane dressings for radiotherapy-induced skin damage.

Radiotherapy is one of the mainline treatments for cancer. One of the side effects associated with radiotherapy includes skin problems, which range from mild (dull erythema and tightening of the skin) to severe (moist desquamation resulting in open wounds that can be very painful associated with sloughy and, in some severe cases, necrosis). The increased use of advanced radical treatments, such as intensity-modulated radiotherapy treatment (IMRT), can also result in a higher number of patients experiencing skin reactions. It is estimated that approximately 87% of patients will experience a moderate-to-severe skin reaction (Harris et al, 2011) An evaluation was undertaken in 20 patients with head and neck cancer following a prescribed treatment of radiotherapy to compare a polymeric membrane dressing (PolyMem®) against the standard treatment. The standard treatment consisted of topical aqueous cream at the start of radiotherapy with the addition of paraffin gauze when moist desquamation occurred. A bespoke evaluation form was completed for a period of 4 weeks or until healed. Patients were asked to complete both qualitative descriptions and numerical scores of pain for symptoms and procedural pain. Analgesia and sleep patterns were logged and, in addition, free text diaries were provided for up to 4 weeks. Common themes were identified and qualitative data analysed.

Anthropological and radiographic comparison of antemortem surgical records for identification of skeletal remains.

This case review illustrates the important contributions of forensic archeological methods and forensic anthropological analysis to the identification of found skeletal remains. After reassociation of skeletal remains found in two locations, anthropological analysis provided the basis for a presumptive identification and a request for antemortem medical records. Partial DNA profiles were supportive but not conclusive and antemortem dental records were not available. Comparison of antemortem traumas, skeletal morphology, and surgical artifacts with antemortem radiographs and surgical records led to positive identification of an individual missing for almost a decade.