Early Blockade of CB1 Receptors Ameliorates Schizophrenia-like Alterations in the Neurodevelopmental MAM Model of Schizophrenia

In agreement with the neurodevelopmental hypothesis of schizophrenia, prenatal exposure of Sprague-Dawley rats to the antimitotic agent methylazoxymethanol acetate (MAM) at gestational day 17 produces long-lasting behavioral alterations such as social withdrawal and cognitive impairment in adulthood, mimicking a schizophrenia-like phenotype. These abnormalities were preceded at neonatal age both by the delayed appearance of neonatal reflexes, an index of impaired brain maturation, and by higher 2-arachidonoylglycerol (2-AG) brain levels. Schizophrenia-like deficits were reversed by early treatment [from postnatal day (PND) 2 to PND 8] with the CB1 antagonist/inverse agonist AM251 (0.5 mg/kg/day). By contrast, early CB1 blockade affected the behavioral performance of control rats which was paralleled by enhanced 2-AG content in the prefrontal cortex (PFC). These results suggest that prenatal MAM insult leads to premorbid anomalies at neonatal age via altered tone of the endocannabinoid system, which may be considered as an early marker preceding the development of schizophrenia-like alterations in adulthood.

Advances of Benzimidazole Derivatives as Anticancer Agents: Bench to Bedside

Benzimidazole is one of the privileged nitrogen-containing scaffolds known for its versatile diversified role in insecticides, pesticides, dyes, pigments and pharmaceuticals. Due to its electron-rich environment, structural features and binding potency of various therapeutic targets, benzimidazole derivatives exhibit a broad spectrum of biological activity that majorly includes antimicrobial, antifungal, analgesics, anti-diabetic and anticancer agents. Several benzimidazole scaffolds bearing drugs are clinically approved; they are used for various indications. For example, Bilastine, Lerisetron, Maribavir and Nocodazole are the most widely used benzimidazole-based marketed drugs available as an antihistamine, antiviral and antimitotic agent, respectively. Another example is the recently approved anticancer drug Binimetinib and Selumetinib, which are indicated for BRAF mutated melanoma and plexiform neurofibromas. Not only this, many benzimidazole-based anticancer drugs are in late phases of clinical development. Due to the vast therapeutic potential of benzimidazole scaffold in cancer research, medicinal chemists have gained a lot of attraction to explore it more and develop novel, highly effective and target-specific benzimidazole-based potential anticancer drugs.

Identification of the ploidy level of plants in grape breeding

Полиплоидные формы обращают на себя внимание своими положительными свойствами, одним из которых является увеличение по сравнению с диплоидными сортами размеров хозяйственно ценных органов. Значительное количество работ по полиплоидии растений в последнее время обусловлено развитием аналитических методов, таких как современные способы приготовления цитогенетических препаратов, цифровая микроскопия, проточная цитометрия, ПЦР-анализ. В настоящее время для получения полиплоидных форм растений используют системы культуры ткани. Успех индукции полиплоидии зависит от различных факторов: состава питательной среды, антимитотического агента, типа эксплантов, времени воздействия и концентрации веществ. В программах создания полиплоидных форм растений проводят исследования с использованием прямого подсчета хромосом, проточной цитометрии, ПЦР-анализа, а также косвенных методов изучения морфологических особенностей объектов. Методы изучения структуры эпидермиса листа являются простыми, быстрыми, неразрушающими и не требующими дорогих реагентов или оборудования. В качестве морфологических индикаторов плоидности обычно используют параметры устьиц (частота устьиц, размеры замыкающих клеток и количество хлоропластов в устьицах). Предлагаются простые протоколы прямого и косвенного методов анализа плоидности винограда. Наиболее успешными работами в области изучения плоидности растений можно считать исследования комплексные. Косвенные методы анализа следует использовать для массового скрининга исходной выборки, прямые методы - для точного изучения генома отобранных растений. Изучение морфологических особенностей эпидермиса листьев может быть использовано в селекционных программах создания виноградных полиплоидов. Исследование дает рациональное обоснование дальнейшей работы по анализу цитогенетических и морфологических особенностей полиплоидных растений винограда. Polyploid forms attract attention for their positive properties, one of which is an increase in the size of economically valuable organs compared to diploid varieties. A significant number of works on plant polyploidy in recent years is due to the development of analytical methods, such as modern methods for the preparation of microslides, digital microscopy, flow cytometry, PCR- analysis. Tissue culture systems are currently used to obtain polyploid forms of plants. The success of polyploidy induction depends on various factors, such as composition of the nutrient medium, antimitotic agent, type of explants, time of exposure, and concentration of substances. In programs for creating polyploid forms of plants, the research is carried out using direct chromosome counting, flow cytometry, PCR-analysis, as well as indirect methods for studying morphological characteristics of objects. Methods for study the structure of leaf epidermis are simple, fast, non-destructive and not requiring expensive reagents or equipment. Stomatal parameters (stomatal density, guard cell size, and the number of chloroplasts in stoma) are commonly used as morphological indicators of ploidy. Simple protocols of direct and indirect methods of ploidy analysis for grapes are proposed. Complex research can be considered as the most successful in the field of plant ploidy studies. Indirect methods of analysis should be used for mass screening of the initial sample, direct methods - for precise study of the genome of selected plants. The study of morphological characteristics of leaf epidermis can be used in breeding programs for the creation of grape polyploids. The research provides a rational basis for further work on the analysis of cytogenetic and morphological features of polyploid grape plants.

Purification and In Vitro Evaluation of an Anti-HER2 Affibody-Monomethyl Auristatin E Conjugate in HER2-Positive Cancer Cells

A promising approach for the development of high-affinity tumor targeting ADCs is the use of engineered protein drugs, such as affibody molecules, which represent a valuable alternative to monoclonal antibodies (mAbs) in cancer-targeted therapy. We developed a method for a more efficient purification of the ZHER2:2891DCS affibody conjugated with the cytotoxic antimitotic agent auristatin E (MMAE), and its efficacy was tested in vitro on cell viability, proliferation, migration, and apoptosis. The effects of ZHER2:2891DCS-MMAE were compared with the clinically approved monoclonal antibody trastuzumab (Herceptin®). To demonstrate that ZHER2:2891DCS-MMAE can selectively target HER2 overexpressing tumor cells, we used three different cell lines: the human adenocarcinoma cell lines SK-BR-3 and ZR-75-1, both overexpressing HER2, and the triple-negative breast cancer cell line MDA-MB-231. MTT assay showed that ZHER2:2891DCS-MMAE induces a significant time-dependent toxic effect in SK-BR-3 cells. A 30% reduction of cell viability was already found after 10 min exposure at a concentration of 7 nM (IC50 of 80.2 nM). On the contrary, MDA-MB-231 cells, which express basal levels of HER2, were not affected by the conjugate. The cytotoxic effect of the ZHER2:2891DCS-MMAE was confirmed by measuring apoptosis by flow cytometry. In SK-BR-3 cells, increasing concentrations of conjugated affibody induced cell death starting from 10 min of treatment, with the strongest effect observed after 48 h. Overall, these results demonstrate that the ADC, formed by the anti-HER2 affibody conjugated to monomethyl auristatin E, efficiently interacts with high affinity with HER2 positive cancer cells in vitro, allowing the selective and specific delivery of the cytotoxic payload.

In vivo polyploidy induction of Phalaenopsis amabilis in a bubble bioreactor system using colchicine

Phalaenopsis amabilis Blume var. grandiflora Bateman is economically important as cut and pot flower. Polyploidy is considered as a valuable tool in improvement and evolution of ornamental plants. Protocorm-like bodies (PLBs) of P. amabilis were cultured on Murashige and Skoog medium containing 0.20 mg L-1 IBA together with 2.00 mg L-1 KIN and 1.00 g L-1 activated charcoal and grown for a period of five months. Fully-developed plantlets from in vitro grown PLBs were immersed in a bubble reactor filled with half-strength Hoagland solution containing the antimitotic agent colchicine (0.05%, 0.10% and 0.15%, w/v) for 72 h with a few drops (1 mL of 0.1%) of octylphenoxypolyethoxyethanol or Nonidet (P-40) as a surfactant. Plantlets were aerated to prevent hypoxia. Colchicine-treated and untreated plantlets were transferred to pots for a period of 60 days. Tetraploidy was successfully induced by 0.15% colchicine. Polyploidy levels were firstly detected using flow cytometry and then confirmed by cytological and morphological observations. The chromosome number was 2n = 2x = 38 in diploids and 2n = 4x = 76 in tetraploid. Incubation of plantlets in liquid medium containing 0.15% colchicine induced the maximum recovered tetraploids with minimum frequency of survival (50%). The tetraploid plants were more compact and exhibited round and thick leaves with darker green color than diploids. Stomata size in tetraploids were larger with less density than diploids. Chloroplast number in guard cells of tetraploids was about two times more than that of control. These results indicate that induction of tetraploids are a reliable and powerful tool for generation of novel phenotypes with ornamental and horticultural value for genetic improvement and breeding. Produced tetraploids in current study have potential in the ornamental/floriculture trade.

In Vitro Polyploidization of Thymus vulgaris L. and Its Effect on Composition of Essential Oils

The aim of this work was to find an effective protocol for in vitro propagation and to perform the in vitro polyploidization of diploid Thymus vulgaris (2n = 30) using two experimental methods based on the use of oryzalin, an antimitotic agent. The ploidy level of the obtained shoots was checked by flow cytometric analysis. The most efficient conditions for inducing polyploidy were oryzalin concentrations of 0.346 and 1.73 mg L−1 present in the medium for two weeks. The vital polyploid shoots were multiplied for further evaluation, rooting and final transfer to nonsterile glasshouse and field conditions. The chemical compositions of the essential oils (EOs)—which were obtained from dried field grown plants by steam distillation—were analyzed by gas chromatography/mass spectrometry (GC/MS). The identified substances contributed approximately 95% to the total peak area. Statistical analysis revealed that the tetraploid subclone and the diploid reference plant do not differ in total terpene content, but they do differ in the relative proportions of all the individual terpenes with the exception of α-pinene and UN5, indicating that both clones produce EOs of different quality. The obtained results showed the possibility of developing more efficient botanical insecticides based on EOs obtained from the tetraploid plants.

Cutaneous Mucormycosis: Case Report

Treatment with 5-FU cream, an antimitotic agent, is primarily used for the treatment of superficial keratinocytic lesions. This treatment modality has the potential to cause severe localized inflammation with subsequent erythema, pain and crusted erosions. Cutaneous mucormycosis is an opportunistic emerging fungal infection. It is rapidly progressive and affects immunosuppressed or poorly controlled diabetic patients primarily by direct inoculation or secondarily by dissemination. The clinical presentation can be challenging due to its initial nonspecific features and therefore leading to a delay in the diagnosis. Herein, we report a case of mucormycosis of the scalp following treatment with topical 5-FU.

Effects of Ferrocenyl 4-(Imino)-1,4-Dihydro-quinolines on Xenopus laevis Prophase I - Arrested Oocytes: Survival and Hormonal-Induced M-Phase Entry

Xenopus oocytes were used as cellular and molecular sentinels to assess the effects of a new class of organometallic compounds called ferrocenyl dihydroquinolines that have been developed as potential anti-cancer agents. One ferrocenyl dihydroquinoline compound exerted deleterious effects on oocyte survival after 48 h of incubation at 100 μM. Two ferrocenyl dihydroquinoline compounds had an inhibitory effect on the resumption of progesterone induced oocyte meiosis, compared to controls without ferrocenyl groups. In these inhibited oocytes, no MPF (Cdk1/cyclin B) activity was detected by western blot analysis as shown by the lack of phosphorylation of histone H3. The dephosphorylation of the inhibitory Y15 residue of Cdk1 occurred but cyclin B was degraded. Moreover, two apoptotic death markers, the active caspase 3 and the phosphorylated histone H2, were detected. Only 7-chloro-1-ferrocenylmethyl-4-(phenylylimino)-1,4-dihydroquinoline (8) did not show any toxicity and allowed the assembly of a histologically normal metaphase II meiotic spindle while inhibiting the proliferation of cancer cell lines with a low IC50, suggesting that this compound appears suitable as an antimitotic agent.

Effects of Electrical Stimulation on Peripheral Nerve Regeneration in a Silicone Rubber Conduit in Taxol-Treated Rats

Taxol, a type of antimitotic agent, could modulate local inflammatory conditions in peripheral nerves, which may impair their regeneration and recovery when injured. This study provided in vivo trials of silicone rubber chambers to bridge a long 10 mm sciatic nerve defect in taxol-treated rats. It was aimed to determine the effects of electrical stimulation at various frequencies on regeneration of the sciatic nerves in the bridging conduits. Taxol-treated rats were divided into four groups (n = 10/group): sham control (no current delivered from the stimulator); and electrical stimulation (3 times/week for 3 weeks at 2, 20, and 200 Hz with 1 mA current intensity). Neuronal electrophysiology, animal behavior, neuronal connectivity, macrophage infiltration, calcitonin gene-related peptide (CGRP) expression levels, and morphological observations were evaluated. At the end of 4 weeks, animals in the low- (2 Hz) and medium-frequency (20 Hz) groups had dramatic higher rates of successful regeneration (90% and 80%) across the wide gap as compared to the groups of sham and high-frequency (200 Hz) (60% and 50%). In addition, the 2 Hz group had significantly larger amplitudes and evoked muscle action potentials compared to the sham and the 200 Hz group, respectively (P < 0.05). Heat, cold plate licking latencies, motor coordination, and neuronal connectivity were unaffected by the electrical stimulation. Macrophage density, CGRP expression level, and axon number were all significantly increased in the 20 Hz group compared to the sham group (P < 0.05). This study suggested that low- (2 Hz) to medium-frequency (20 Hz) electrical stimulation could ameliorate local inflammatory conditions to augment recovery of regenerating nerves by accelerating their regrowth and improving electrophysiological function in taxol-treated peripheral nerve injury repaired with the silicone rubber conduit.