The application of gene silencing in proteomics: from laboratory to clinic

Introduction: Since the completion of genome sequencing, gene silencing technologies have emerged as powerful tools to study gene functions in various biological processes, both in vivo and in vitro. Moreover, they have also been proposed as therapeutic agents to inhibit selected genes in a variety of pathological conditions, such as cancer, neurodegenerative, and cardiovascular diseases.

Area covered: This review summarizes the mechanisms of action and applications of genome editing tools, from RNA interference to clustered regularly interspaced short palindromic repeats-based systems, in research and in clinics. We describe their essential role in high-throughput genetic screens and, in particular, in functional proteomics studies, to identify diagnostic markers and therapeutic targets. Indeed, gene silencing and proteomics have been extensively integrated to study global proteome changes, posttranslational modifications, and protein–protein interactions.

Expert commentary: Functional proteomics approaches that leverage gene silencing tools have been successfully applied to examine the role of several genes in various contexts, leading to a deeper knowledge of biological pathways and disease mechanisms. Recent developments of gene silencing tools have improved their performance, also in terms of off-targets effects reduction, paving the way for a wider therapeutic application of these systems.     

Estimation of fetal weight by ultrasound.

Estimation of fetal size by ultrasound has supported two recent concepts of fetal growth. Firstly, the normal weight gain is constant from 28 weeks of gestation until several weeks after term; about 27 g/day. Secondly, the average weight of fetuses born preterm is lower than the average weight of fetuses of the same gestational age who remain in utero. This means that up to 40% of infants born at 28-30 weeks of gestation are small-for-gestational age, when related to the biological optimum. Unfortunately, the test-retest variation of fetal weight estimation is as much as 50% of the standard deviation of the gestational-age-specific fetal weight distribution. This means that fetal growth has to be followed for many weeks to document statistically significant deviations. No study has, as yet, demonstrated the value of this approach to distinguish between intrauterine growth retardation and growth along low centiles by fetuses of reduced growth potential.     

基因工程抗体的研究进展

基因工程抗体以其独特的优点（免疫原性低、可按人的意愿加以改造等）正逐渐取代动物源性单抗。随着基因工程和蛋白质工程等生物技术在抗体研制领域的广泛应用,适应不同需要的基因工程抗体的种类日趋多样化,构建日趋合理化,在体内的生物学效应与日臻完善,使之较天然单抗的治疗效果更好,范围更广,并在初步临床试用中展示了光辉的前景。     

The future of organ transplantation: from the laboratory to the clinic

This is a short review of tolerance from the point of view of the clinician. Various examples of tolerance occurring in patients and animal models that relate to the clinical experience are described. It is suggested that there may be different mechanisms by which tolerance is achieved, but from the patient's point of view operational tolerance is the goal, whereby, after a short induction procedure, the patient will maintain good function in the grafted organ indefinitely without maintenance immunosuppression. It is pointed out that such a goal may be difficult to achieve with any given protocol due to the enormous variation between donors and recipients of organ grafts of tissue matching, innate immune reactivity and susceptibility to disturbance of a tolerant state by infections or allergic reactions. Thus the case is made for prope or almost tolerance in which graft acceptance is maintained by a low, non-toxic dosage of maintenance immunosuppression that may not be required indefinitely.     

Effect of tolerance to paternal antigens on placental and fetal weight in the mouse

Placental and fetal weight was measured on the 16th day of pregnancy in normal and tolerant mice. Placental weight was increased in A mice tolerant to C57BL/10 mice only when donor cells from males were used. Fetal weight was unaffected.     

Estimation of gestational age and assessment of canine fetal maturation using radiology and ultrasonography: a review

Since the duration of pregnancy in the bitch is relatively short, it is critical that fetuses are fully mature prior to delivery for them to survive. For breeders to be able to prepare for normal whelpings and align medical care in case of emergency, an estimated due date is necessary. In cases where ovulation timing is lacking and there is a singleton fetus or oversize fetuses, it is necessary to ascertain gestational age prior to setting the date of Cesarean section. In high-risk pregnancies, where there is poor or no ovulation timing, determination of fetal maturation and gestational age will assist in determining if pregnancy has progressed long enough to allow delivery of viable puppies. In cases where bitches are receiving supplemental progesterone for pregnancy maintenance medications must be discontinued at an appropriate time to permit delivery of viable puppies. It also allows for estimation of the likelihood of fetal survival if the pregnancy is terminated due to failing bitch health, with subsequent surgical delivery of the fetuses. Use of breeding dates alone does not provide due dates with adequate accuracy. In cases where there has been inadequate or no breeding management or ovulation timing, estimation of due date can be performed at the time of pregnancy diagnosis, or closer to term. Radiography can be used to confirm pregnancy and facilitate determination of gestational age, beginning 45d after the LH surge. Ultrasonography can be used from 19 to 21d after the LH surge to term to confirm pregnancy and predict gestational age, and from 25 or 26d to term to assess fetal viability and fetal stress.     

Primate stem cells: bridge the translation from basic research to clinic application

A growing body of literature has shown that stem cells are very effective for the treatment of degenerative diseases in rodents but these exciting results have not translated to clinical practice. The difference results from the divergence in genetic, metabolic, and physiological phenotypes between rodents and humans. The high degree of similarity between non-human primates(NHPs) and humans provides the most accurate models for preclinical studies of stem cell therapy. Using a NHP model to understand the following key issues, which cannot be addressed in humans or rodents, will be helpful for extending stem cell applications in the basic science and the clinic. These issues include pluripotency of primate stem cells, the safety and efficiency of stem cell therapy, and transplantation procedures of stem cells suitable for clinical translation. Here we review studies of the above issues in NHPs and current challenges of stem cell applications in both basic science and clinical therapies. We propose that the use of NHP models, in particular combining the serial production and transplantation procedures of stem cells is the most useful for preclinical studies designed to overcome these challenges.     

Birth weight from pregnancies dated by ultrasonography in a multicultural British population.

OBJECTIVE--To produce standard curves of birth weight according to gestational age validated by ultrasonography in the British population, with particular reference to the effects of ethnic origin. DESIGN--Retrospective analysis of computerised obstetric database. SETTING--Three large maternity units associated with Nottingham University with over 16,000 deliveries a year. PATIENTS--41,718 women with ultrasound dated singleton pregnancies and delivery between 168 and 300 days'' gestation. MAIN OUTCOME MEASURES--Length of gestation, ethnic origin, parity, maternal height and weight at booking, smoking during pregnancy; the effect of these variables on birth weight. RESULTS--Birth weights from ultrasound dated pregnancies have a higher population mean and show less flattening of the birthweight curve at term than those of pregnancies dated from menstrual history. Significant differences were observed in mean birth weights of babies of mothers of European origin (3357 g), of Afro-Caribbean origin (3173 g), and from the Indian subcontinent (3096 g). There were also significant interethnic differences in length of gestation, parity, maternal height, booking weight, and smoking habit which affected birth weight. The ethnic differences in birth weight were even greater when the effect of smoking was excluded. CONCLUSIONS--Birthweight standards require precise dating of pregnancy and should describe the population from which they were derived. In a heterogeneous maternity population the accurate assessment of an individual baby''s weight needs to take the factors which affect birthweight standards into consideration.     

CAMPATH: from concept to clinic

Lymphocyte depletion has a long history in the area of therapeutic immunosuppression. CAMPATH-1H (alemtuzumab) was generated in an attempt to replace anti-lymphocyte globulins in the transplant arena. Its efficacy in killing lymphocytes has established it as a licensed drug for the management of chronic lymphocyte leukaemia. Short-term therapy with alemtuzumab has demonstrated long-term benefit in a number of autoimmune conditions. This drug has the potential to facilitate recruitment of tolerance processes so enabling drug minimization in transplantation, autoimmune and hypersensitivity diseases.     

Toxicity of baited spinosad formulations to Ceratitis capitata: from the laboratory to the application

GF‐120, a fruit fly bait designed to attract and kill adult fruit flies, was tested in the laboratory and outdoors to determine effects of pre‐treatment diet and bait aging on mortality of Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). Two spinosad‐based compounds, GF‐120 and Tracer^® Ultra, had generated two distinctive dose–mortality responds, with LC⁸⁰, LC⁹⁰, and LC⁹⁹ values of 2.4, 2.8, and 4.1 p.p.m., and 255, 479, and 1 143 p.p.m., respectively. The residues of GF‐120 drops, after feeding to the flies, generated 14.3% mortality. The droplet size of the baited spray plays an important role. The toxicity of large drops lasted more than that of small droplets. In the field, exposure to the sun further deteriorates the compound, which lost 50% of its toxicity within 6 days. Disappearance of the compound in the field, due to consumption by various insects, also played a role as 50% of the GF‐120 drops disappeared within 7 days. As mortality was directly related to the amount of insecticide eaten, the effect of GF‐120 depended on the feeding status of the flies: well‐fed flies were almost unaffected compared with starved ones.     

From nature to the laboratory and into the clinic

Natural products possess a broad diversity of structure and function, and they provide inspiration for chemistry, biology, and medicine. In this review article, we highlight and place in context our laboratory’s total syntheses of, and related studies on, complex secondary metabolites that were clinically important drugs, or have since been developed into useful medicines, namely amphotericin B (1), calicheamicin γ₁^I (2), rapamycin (3), Taxol^® (4), the epothilones [e.g., epothilones A (5) and B (6)], and vancomycin (7). We also briefly highlight our research with other selected inspirational natural products possessing interesting biological activities [i.e., dynemicin A (8), uncialamycin (9), eleutherobin (10), sarcodictyin A (11), azaspiracid-1 (12), thiostrepton (13), abyssomicin C (14), platensimycin (15), platencin (16), and palmerolide A (17)].     

Interlitter variability in fetal body weight in mouse offspring from continuous, overnight, and short-period matings

When female mice of the Jcl:ICR strain were mated with a male either (1) continuously throughout the day or (2) overnight or (3) during 2 hours in the morning at light period, the interlitter variability of fetal body weight on the 18th day of gestation was smallest in the group with short-period mating. Thus, in the embryonic stage-specific teratological experiments, this mating schedule is advised. Even for routine reproductive toxicity testing protocols, the short-period mating may be preferable for the purpose of increasing test sensitivity.     

Catecholamine-induced stimulation of testosterone production by Leydig cells from fetal mouse testis

In contrast to the strong stimulation of testosterone production by hCG, L-isoproterenol had little effect on freshly isolated Leydig cells from 18-day-old mouse fetuses. However, the ability of fetal Leydig cells to respond to L-isoproterenol exposure increased during culture (0-24 h). The response of the cultured cells to L-isoproterenol was dose-dependent with an ED50 at 2 X 10(-7) M. Adrenaline and noradrenaline at a concentration of 10(-5) M also increased testosterone production by cultured fetal Leydig cells. DL-Propranolol, a beta-antagonist, inhibited L-isoproterenol-stimulated testosterone production in a dose-dependent manner, while phentolamine, an alpha-adrenergic antagonist, had no effect. These results suggest that catecholamines may play an essential role in the control of testicular steroidogenesis during fetal development.