Advancements in Graft Placement Techniques.

For decades, the placement of follicular units (FUs) into incisions in the recipient area was exclusively carried out using forceps. In 1992, Dr. Choi introduced an instrument known as the "implanter," which had the advantage of simultaneously creating incisions and placing FUs without damaging sensitive parts. Its initial popularity was greater in the East, primarily due to the characteristics of Asian hair. Asian hair is typically straight and thick, with FUs mostly consisting of just one or two hair.With the description of the follicular unit excision (FUE) technique in 2002 and its widespread adoption nearly a decade later, the advantages of using the implanter also gained popularity in the West. The uniformity in the size of FUs provided by the FUE technique and the possibility of delegating the placement were key attractions in the use of this placement tool. In addition to the traditional Korean implanter with a sharp needle, other implanters and inserters have been described.The choice of implantation technique depends on individual adaptation and the advantages and disadvantages offered by each instrument. Although forceps allow for safe placement in the hands of well-trained teams, the increased fragility of FUs obtained with the FUE technique has led to the growing acceptance of techniques that employ implanters and inserters.

Whole-body autoradiography in drug discovery.

In the drug discovery process the pharmacokinetic screening, drug stability studies, evaluation of metabolites, CYP involvement, enzyme induction and inhibition, and excretion studies play a major role. The use of more sensitive and novel detection systems have made the discovery process less cumbersome than in previous years. In particular, the use of whole-body autoradiography (WBA) for tissue distribution, which was once considered an impractical tool, owing to the long turn around time (4-10 weeks), is coming to the forefront for rapidly resolving issues encountered in discovery. In today's research environment early lead compounds can be radio-labeled and whole-body sections imaged quickly (3-5 days) using new techniques, which has made (14)C- and (3)H-WBA a viable tool. The technique has been used in vivo in species from mice to monkeys, and ex vivo and/or in vitro in larger animals and humans. WBA has considerable merit in identifying "pharmacodeficient" compounds and providing insight on mechanistic questions. WBA data can provide information related to tissue pharmacokinetics, routes of elimination, CYP or Pgp mediated drug-drug interactions, tissue distribution, site specific drug localization and retention, metabolism, clearance, compound solubility issues, routes of administration, penetration into specific targets (e.g., tumors), tissue binding (e.g., melanin), and interspecies kinetics. Thus, WBA is quickly becoming part of the battery of studies conducted during the lead optimization process to select optimal drug candidates. Examples of the use of the WBA tool in early discovery are reviewed.

Quantitative whole-body autoradiography in the pharmaceutical industry. Survey results on study design, methods, and regulatory compliance.

INTRODUCTION: Quantitative whole-body autoradiography (QWBA) is a technique used to determine the tissue distribution of radiolabeled compounds in laboratory animals. This relatively new technique is quickly replacing wet-tissue dissection techniques, which, up to now, have been used by the pharmaceutical industry when performing tissue distribution studies to develop new drugs and to address regulatory compliance needs. In an effort to harmonize QWBA procedures across the pharmaceutical industry, the Society for Whole Body Autoradiography (SWBA) surveyed its membership to gain insight into the procedures and practices being used to perform tissue distribution studies conducted in support of drug development. METHODS: The survey polled 29 respondents, who represent pharmaceutical companies in the United States, Europe, and Asia. Participants answered approximately 50 questions related to study design, applications, autoradiography methods, tissue quantitation, and regulatory compliance. RESULTS: The survey revealed general consistencies and inconsistencies among the labs that responded. Consistencies were related to: isotope use and doses of radioactivity, number of animals per time point, exsanguination of animals, freezing methods, section thickness, tissue collection lists, section lyophilization, imaging technology, blood and calibration standards, tissues and sections sampled for quantitation, use of QWBA data for human dosimetry, and QWBA method validation. Inconsistencies were related to: number of time points used, euthanasia methods, carcass freezing time, microtome calibration, section thickness verification, sample collection, validation of commercial standards, use of background measurements during calibration, definition of limits of quantitation, reporting of extrapolated values, reexposure of section to determine low levels, computer system validation, definitions of raw data, audit trail documentation, studies performed under Good Laboratory Practices (GLP) vs. non-GLP conditions. DISCUSSION: The survey indicated that most labs are now using QWBA to perform their tissue distribution studies and that these data have been submitted and accepted by regulatory authorities around the world. Procedures and practices involved in the design of these studies appear to vary somewhat. An important inconsistency found related to the number of time points used to determine the pharmacokinetic (PK) parameters for tissues, which may effect the reliability of these parameters for use in predicting human exposure to radioactivity during human radiolabeled studies. Survey results regarding QWBA methods indicated that there is a lot of consistency across surveyed labs; however, there are some inconsistent areas that raise regulatory compliance issues and these are related to the verification of section thickness, validation of commercial standards and their use in quantitation, definitions of limits of quantitation, and consideration of background measurements during quantitation. This survey provides autoradiographers, managers, and regulators with an important reference on the state-of-the art of QWBA and shows that the technique has gained wide acceptance across the pharmaceutical industry. However, it also shows that there are some key areas, such as inconsistencies in the procedures used for quantitation, that investigators may want to probe further to assure that the highest quality and most useful studies are performed.

Methods determining phosphor imaging limits of quantitation in whole-body autoradiography rodent tissue distribution studies affect predictions of 14C human dosimetry.

INTRODUCTION: Radioluminography, or phosphor imaging, is often used in rodent quantitative whole-body autoradiography (QWBA) studies to determine the tissue distribution and pharmacokinetic (PK) parameters of new pharmaceutical entities in rodents. The rodent tissue pharmacokinetics information are then used to predict human radiation exposure to 14C or 3H during human radioisotope mass balance studies. The human dosimetry estimation can be biased by the method used to determine the lower limit of quantitation (LOQ) of the phosphor imager. A survey of autoradiographers revealed that at least five different methods are used to determine phosphor imager LOQ. The objective of this study is to compare and evaluate the human dosimetry estimates obtained by applying those five LOQ methods to a single set of WBA data. METHODS: Five different phosphor imager LOQ determination methods were applied to a single set of QWBA rodent tissue distribution data to produce five tissue concentration time profiles. Tissue PK parameters were determined for each profile and subsequently used to calculate the 14C exposure for a proposed human 14C mass balance study. RESULTS: A threefold difference was observed among the five predictions of human 14C exposure when the five different phosphor imager LOQ values were applied to the initial data set. DISCUSSION: The method chosen to determine the phosphor imaging LOQ for QWBA rodent tissue distribution study could impact the human 14C exposure estimates. The end result may either under- or overestimate the 14C-tissue exposure in humans during radioisotope studies, depending on the method used to determine LOQ. We recommend two approaches to reduce the variations in the determination of rodent tissue distribution pharmacokinetics: (1) Set more sampling time points to cover the terminal phase to obtain more accurate t 1/2 and (2) use Method 3 or the small sized sampling tool of Method 5 for LOQ determinations because it is a balanced approach for both simplicity and accuracy.