Spotlight on Volasertib: Preclinical and Clinical Evaluation of a Promising Plk1 Inhibitor.

Considering the important side effects of conventional microtubule targeting agents, more and more research focuses on regulatory proteins for the development of mitosis-specific agents. Polo-like kinase 1 (Plk1), a master regulator of several cell cycle events, has arisen as an intriguing target in this research field. The observed overexpression of Plk1 in a broad range of human malignancies has given rise to the development of several potent and specific small molecule inhibitors targeting the kinase. In this review, we focus on volasertib (BI6727), the lead agent in category of Plk1 inhibitors at the moment. Numerous preclinical experiments have demonstrated that BI6727 is highly active across a variety of carcinoma cell lines, and the inhibitor has been reported to induce tumor regression in several xenograft models. Moreover, volasertib has shown clinical efficacy in multiple tumor types. As a result, Food and Drug Administration (FDA) has recently awarded volasertib the Breakthrough Therapy status after significant benefit was observed in acute myeloid leukemia (AML) patients treated with the Plk1 inhibitor. Here, we discuss both preclinical and clinical data available for volasertib administered as monotherapy or in combination with other anticancer therapies in a broad range of tumor types.

BMI-related metabolic composition of the follicular fluid of women undergoing assisted reproductive treatment and the consequences for oocyte and embryo quality.

STUDY QUESTION: Is the metabolic composition of the follicular fluid of women undergoing assisted reproductive treatment (ART) related to serum composition and BMI and is it associated with oocyte and embryo quality? SUMMARY ANSWER: We showed that metabolic alterations in the serum are reflected in the follicular fluid and that some of these alterations may affect oocyte quality, irrespective of BMI. WHAT IS KNOWN ALREADY: Many studies have focused on the effect of metabolic disorders, such as obesity and type 2 diabetes, on assisted reproduction outcomes. There are, however, only few studies focusing on the importance of the correlation between serum and follicular fluid compositions and the composition of the follicular fluid as the oocyte's micro-environment, affecting its development and subsequent embryo quality. DESIGN, PARTICIPANTS AND SETTING: In this prospective cohort study, patient information, fertility treatment outcome data, follicular fluid and serum were obtained from women undergoing ART. Patients were categorized according to their BMI (kg/m(2)) as normal (n = 60), overweight (n = 26) or obese (n = 20). Serum and follicular fluid samples were analyzed for urea, total protein, albumin, cholesterol, high-density lipoprotein cholesterol, triglycerides, non-esterified fatty acids, apolipoprotein A1, apolipoprotein B, glucose, lactate, C-reactive protein, insulin-like growth factor -1 (IGF-1), IGF-binding protein 3 (only in follicular fluid), free carnitine and total carnitine. Metabolite concentrations in serum and follicular fluid samples were correlated and were associated with BMI and fertility treatment outcome. MAIN RESULTS: Most serum metabolite differences between patients were reflected in the follicular fluid (P < 0.05). Follicular fluid apolipoprotein A1 and follicular fluid total protein concentrations negatively affected oocyte quality parameters (P < 0.05). However, overall BMI-related associations were poor. BIAS, CONFOUNDING AND OTHER REASONS FOR CAUTION: In this study, we included every patient willing to participate. Within this cohort, women with a BMI transcending 35 kg/m(2) were scarce (n = 2), because extremely overweight women are mostly advised to lose weight before starting ART. Furthermore, the number of patients in each BMI group was different, possibly masking associations between the metabolic composition of serum and follicular fluid and oocyte quality parameters. GENERALIZABILITY TO OTHER POPULATIONS: There were significant associations indicating that metabolic changes in the serum are reflected in the follicular fluid, potentially affecting oocyte quality, irrespective of the patient's BMI. For ethical reasons, this study only focused on women already in need of artificial reproductive treatment. From a metabolic point of view, we consider this cohort as a representative sample of all women of reproductive age. STUDY FUNDING: This study was funded by the special research fund, university of Antwerp (BOF UA). None of the authors has any conflict of interest to declare.

Exploring the design of a lightweight, sustainable and comfortable aircraft seat.

BACKGROUND: Making a lightweight seat that is also comfortable can be contradictory because usually comfort improvement means adding a feature (e.g. headrest, adjustable lumbar support, movable armrests, integrated massage systems, etc.), which makes seats heavier. OBJECTIVE: This paper explores the design of an economy class aircraft seat that aims to be lightweight, comfortable and sustainable. METHODS: Theory about comfort in seats, ergonomics, lightweight design, Biomimicry and Cradle to cradle was studied and resulted in a list of requirements that the new seat should satisfy. RESULTS: The design process resulted in a new seat that is 36% lighter than the reference seat, which showed that a significant weight reduction can be achieved. This was completed by re-designing the backrest and seat pan and integrating their functions into a reduced number of parts. Apart from the weight reduction that helps in reducing the airplane's environmental impact, the seat also satisfies most of the other sustainability requirements such as the use of recyclable materials, design for disassembly, easy to repair. A user test compared the new seat with a premium economy class aircraft seat and the level of comfort was similar. CONCLUSIONS: Strong points of the new design were identified such as the lumbar support and the cushioning material, as well as shortcomings on which the seat needs to be improved, like the seat pan length and the first impression. Long term comfort tests are still needed as the seat is meant for long-haul flights.

Preclinical and clinical studies on afatinib in monotherapy and in combination regimens: Potential impact in colorectal cancer.

Targeting the epidermal growth factor receptor (EGFR) with monoclonal antibodies (mAbs) or tyrosine kinase inhibitors (TKI) has been an interesting therapeutic strategy because aberrant activation of this receptor plays an important role in the tumorgenesis of many cancer types, including colorectal cancer (CRC). After the initial promising results of EGFR-targeted therapies, therapeutic resistance is a major clinical problem. In order to overcome resistance to these EGFR-targeted therapies, new treatment options are necessary. In contrast to first generation EGFR inhibitors, afatinib (BIBW2992) is a second-generation irreversible ErbB family blocker that inhibits EGFR as well as HER2 and HER4. Consequently, treatment with afatinib may result in a distinct and more pronounced therapeutic benefit. Preclinical studies have reported promising results for afatinib in monotherapy as well as in combination with other drugs in CRC model systems. Furthermore, clinical studies examining afatinib as single agent and in combination therapy demonstrated manageable safety profile. Nevertheless, only limited antitumor activity has been observed in CRC patients. Although several combination treatments with afatinib have already been investigated, no optimal combination has been identified for CRC patients yet. As molecular tumor characteristics have gained increased importance in the choice of treatment, additional studies with biomarker-driven patient recruitment are required to further explore afatinib efficacy in CRC.

Migration of bovine spermatozoa in a synthetic medium and its relation to in vivo bull fertility.

Although sperm migration has been extensively refined and validated in human infertility studies, its application to predict bovine fertility has been very limited, and a clear relation between the sperm migration distance and in vivo bull fertility has never been demonstrated. A synthetic medium based upon methyl cellulose (MC) was tested for its suitability to serve as a migration medium for frozen-thawed bovine spermatozoa. The effects of the concentration of MC, the incubation time, and sperm concentration on sperm migration capacity was determined. The relation between sperm migration capacity at different incubation times of the frozen-thawed spermatozoa of five bulls, and their 56 days nonreturn rates (NRRs) was assessed in order to evaluate its suitability as a tool to predict in vivo bull fertility. The highest repeatability of the sperm migration test (CV = 10.7%) was obtained when the sperm migration distance of the five vanguard motile spermatozoa was determined at 30 min incubation at 37 degrees C in a migration medium with 1.35% MC. No significant difference in migration distance was demonstrated when sperm concentrations of 100 x 10(6) and 150 x 10(6) spermatozoa/ml, respectively, were used. Despite the relatively high repeatability of the migration test, no relation was found between the sperm migration distance and the 56 days NRRs of five sire bulls. Therefore, the sperm migration test in 1.35% MC cannot be used to predict in vivo bull fertility accurately.

In vitro survival of bovine spermatozoa stored at room temperature under epididymal conditions.

In this study, environmental conditions mimicking those prevailing in the epididymis were used for storing ejaculated bull spermatozoa in vitro during 4 days at ambient temperature. These conditions were low pH, high osmolarity, high sperm concentration and low oxygen tension. Hepes-TALP was used as basic storage medium. Fresh spermatozoa were stored at a concentration of 10 x 10(6)spermatozoa/ml in Hepes-TALP of different pH (pH 4, 5, 6, 7 or 8), and osmolarity (100, 300, 400, 500, 600 or 800 mOsm/kg), and under different atmospheric conditions (nitrogen gassed or aerobic). Spermatozoa were also stored undiluted or at different concentrations: 10x 10(6), 100 x 10(6), 500 x 10(6) or 1 x 10(9)spermatozoa/ml. Sperm parameters such as membrane integrity, motility, mitochondrial membrane potential or DNA fragmentation were used to assess semen quality after storage. Adjustment of the pH of Hepes-TALP to pH 6 yielded significantly better results than storage at all other pH values. Isotonic Hepes-TALP (300 mOsm/kg) had a less detrimental effect on spermatozoa than hypo- and hyperosmotic versions. No differences in sperm parameters were observed when spermatozoa were incubated under aerobic or under nitrogen gassed storage conditions. Optimal sperm concentration in vitro is 10 x 10(6)spermatozoa/ml. This is in contrast with the in vivo situation, where spermatozoa are stored at high concentration. However, better results at high sperm concentrations were obtained when spermatozoa were diluted for less than 5 min in Triladyl-egg yolk-glycerol diluent immediately after ejaculation.

Effect of sperm coating on the survival and penetrating ability of in vitro stored bovine spermatozoa.

The aim of this study was to examine the effect of sperm coating on the survival and penetrating ability of in vitro stored diluted spermatozoa. Bovine semen was collected by means of an artificial vagina connected with a tube containing 5 ml of the commercial Triladyl diluent supplemented with 20% egg yolk and 6.7% glycerol (EYTG). Both EYTG and seminal plasma were removed by centrifugation and the spermatozoa were stored under different in vitro storage conditions. In the first and second experiment, "control" and "coated" spermatozoa were stored in Hepes-TALP (pH 6 and 7) at room temperature. After 4 days of storage, the progressive motility, membrane integrity, mitochondrial membrane potential or DNA integrity of the spermatozoa were evaluated before and after Percoll centrifugation. The in vitro penetration rate of the spermatozoa was examined only after Percoll centrifugation. A significantly (P<0.05) positive influence of sperm coating was observed on the tested sperm characteristics and penetration rate of spermatozoa when they were stored in Hepes-TALP at pH 7, but not at pH 6. In the last experiment, the influence of the storage medium Hepes-TALP (pH 7) or EYTG was investigated on motility, membrane integrity, mitochondrial membrane potential and in vitro penetration potential of "coated" spermatozoa stored at room temperature or at 4 degrees C during 4, 5 and 6 days. After 6 days of storage, a significantly (P<0.05) higher percentage of motile and membrane intact spermatozoa with high mitochondrial membrane potential was obtained in EYTG at both temperatures leading to a significantly higher in vitro penetration rate. These results indicate that sperm coating could preserve sperm characteristics and penetrating capacity of fresh bovine spermatozoa stored in egg yolk containing diluent for up to 6 days.

Storage of fresh bovine semen in a diluent based on the ionic composition of cauda epididymal plasma.

For artificial insemination (AI) in cattle, much lower insemination doses can be applied when fresh semen is used instead of frozen-thawed semen. However, a particular disadvantage of fresh semen is its limited shelf life. As bovine spermatozoa can be stored for several weeks in the cauda epididymis without negative effects on their fertilizing capacity, it is an interesting organ to serve as a model in order to prolong the shelf life of fresh semen. First, the storage capacity of a diluent [cauda epididymal plasma (CEP-1)] with the same ionic composition, pH and osmolarity as the bovine CEP was compared with a Tris diluent for extended preservation of fresh ejaculated bovine semen. Secondly, the ionic composition of the CEP-1 diluent was modified (CEP-2) and its storage capacity was compared with this of the CEP-1 and Tris diluent. Finally, the effect of addition of different polyols (sorbitol, glycerol, mannitol) and egg yolk concentrations (5, 10 and 20%) to the CEP-2 diluent was assessed. Sperm quality decreased rapidly in the CEP-1 diluent. The quality and especially progressive motility of spermatozoa stored in the CEP-2 diluent were better those in the CEP-1 and Tris diluent. No significant effects of different sugars or egg yolk concentrations on the quality of fresh bovine semen in the CEP-2 diluent were observed. In conclusion, the CEP-2 diluent with 10% egg yolk and 1 g/l sorbitol may be used for extended preservation of fresh bovine semen at 5 degrees C up to 6 days.

Function of the cumulus oophorus before and during mammalian fertilization.

CONTENTS: Fertilization encompasses a series of different steps which have to be performed in a well-orchestrated way to create a new individual. They include sperm capacitation, sperm binding and penetration of the zona pellucida, traversing the perivitelline space, binding and fusion with the oolemma, activation of the oocyte and decondensation of the sperm head to form the male pronucleus. In most mammalian species, cumulus cells surround the oocyte at the time of fertilization. Removal of the cumulus oophorus at this point of time often leads to a drop in fertilization rates. It is not yet known how cumulus cells interact with the oocyte or with spermatozoa to promote fertilization. There are different possibilities: 1 cumulus cells cause mechanical entrapment of spermatozoa and guide hyperactivated spermatozoa towards the oocyte, while preventing abnormal spermatozoa to enter the cumulus matrix; 2 cumulus cells create a micro-environment for the spermatozoa which favours their capacitation and penetration into the oocyte; 3 cumulus cells prevent changes in the oocyte which are unfavourable for normal fertilization; these changes can be located in the zona pellucida or in the cytoplasm. In this review, studies in several species are listed to prove the importance of these three cumulus cell functions and the current lines of research are highlighted. Moreover, different ways to improve in vitro fertilization of bovine cumulus-denuded oocytes are discussed.

Lichen planus induced by flunarizine.

A 56-year-old man developed lichen planus while taking flunarizine, a di-fluorinated derivate of cinnarizine. Induction of lichen planus by cinnarizine therapy has been described but the precise etiopathological mechanism is unclear. Although flunarizine is widely prescribed, lichen planus induced (?) by this drug has, to our knowledge, never been reported.

DNA typing of fingerprints using capillary electrophoresis: effect of dactyloscopic powders.

DNA typing is a useful tool in crime solving, not only for blood samples, sperm, or saliva but also for traces of DNA left on tools or pieces of clothing used in burglaries or thefts. On these kinds of samples, the sources of DNA are extremely small amounts of skin debris left after gripping tools. When a sensitive technique such as polymerase chain reaction (PCR) coupled with capillary electrophoresis is used, it is possible to get a profile from these low amounts of DNA. The classic technique in such cases, used in forensic sciences, is to reveal fingerprints by different dactyloscopic powders. Therefore, DNA profiling was performed on physical fingerprints left on glass and wooden plates, in order to establish eventual problems or interferences involved by using both techniques simultaneously. Eleven dactyloscopic powders were investigated on their influence on DNA typing. The results show that some can be used together with DNA profiling but that serious precautions have to be taken to avoid contamination.

Cloning and molecular analysis of two new sesquiterpene cyclases from Artemisia annua L.

Artemisia annua L. is the only source of artemisinin, a new promising antimalarial drug (Qinghaosu Antimalarial Coordinating Research Group, Chin. Med. J. 92 (1979) 811). Our efforts are focused on the overproduction of this valuable medicine by genetic engineered A. annua plants. Therefore, we decided to isolate the gene(s) encoding sesquiterpene cyclase(s) in A. annua as a first step in improving artemisinin yield. Four partial genomic clones, gASC21, gASC22, gASC23 and gASC24, were isolated through polymerase chain reaction (PCR) with degenerated primers based on homologous boxes present in sesquiterpene cyclases from divergent sources. Intron-exon organisation of those partial genomic clones was analysed and it was shown that A. annua contains a gene family for sesquiterpene cyclases. Based on gASC21, gASC22, gASC23 and gASC24 sequences, the full-length cDNA clones cASC34 and cASC125 were subsequently isolated by rapid amplification of cDNA ends PCR. The derived amino acid sequences of both full-length clones show high homology with sesquiterpene cyclases from plants. Reverse transcription-PCR analysis revealed transient and tissue specific expression patterns for cASC34 and cASC125, in contrast to the constitutively expressed 8-epicedrol synthase, a previously reported sesquiterpene cyclase from A. annua. Both cASC34 and cASC125 could only be detected in flowering plants when artemisinin concentration is at highest.