Intrinsic Synergy and Selectivity for the Inhibition of Cancer Cell Growth Generated by a Polymer Ligand of Proximal Enzymes.

A fundamental understanding of the design of polymer ligands of proximal enzymes is essential for the precise targeting of cancer cells, but it is still in its infancy. In this study, we systematically investigated the contribution of the chain length, ligand density, and ligand ratio of proximal enzyme-targeted polymers to the efficacy, synergy, and selectivity for the inhibition of cancer cell proliferation. The results revealed that employing a moderate chain length as a scaffold allowed for an intrinsically high efficacy and synergy of proximal enzyme-targeted polymers, in contrast to single enzyme-targeted polymers that prefer longer chain length for efficacy. The synergy obtained in proximal enzyme targeting was not provided by the combination of the corresponding small molecules. Moreover, the maturation of the synergistic efficacy of the proximal enzyme-targeted polymers also improved selectivity. This study proposes a rational design for polymer inhibitors and/or ligands for cancer cells with a high efficacy and selectivity.

Prevalence of Fragile X-Associated Tremor/Ataxia Syndrome in Patients with Cerebellar Ataxia in Japan.

The presence of fragile X mental retardation 1 (FMR1) premutation has been linked to patients with a certain type of cerebellar ataxia, the fragile X-associated tremor/ataxia syndrome (FXTAS). However, its prevalence in Japan has yet to be clarified. The aim of the present study is to determine the prevalence of FXTAS in Japanese patients with cerebellar ataxia and to describe their clinical characteristics. DNA samples were collected from 1328 Japanese patients with cerebellar ataxia, referred for genetic diagnosis. Among them, 995 patients with negative results for the most common spinocerebellar ataxia subtypes were screened for FMR1 premutation. Comprehensive clinical and radiological analyses were performed for the patients harbouring FMR1 premutation. We herein identified FMR1 premutation from one female and two male patients, who satisfied both clinical and radiological criteria of FXTAS (0.3%; 3/995) as well. Both male patients presented with high signal intensity of corticomedullary junction on diffusion-weighted magnetic resonance imaging, a finding comparable to that of neuronal intranuclear inclusion disease. The female patient mimicked multiple system atrophy in the early stages of her disease and developed aseptic meningitis with a suspected immune-mediated mechanism after the onset of FXTAS, which made her unique. Despite the lower prevalence rate in Japan than the previous reports in other countries, the present study emphasises the necessity to consider FXTAS with undiagnosed ataxia, regardless of men or women, particularly for those cases presenting with similar clinical and radiological findings with multiple system atrophy or neuronal intranuclear inclusion disease.

Fabrication of a Polymeric Inhibitor of Proximal Metabolic Enzymes in Hypoxia for Synergistic Inhibition of Cancer Cell Proliferation, Survival, and Migration.

Since conventional molecular targeted drugs often result in side effects, the development of novel molecular targeted drugs with both high efficacy and selectivity is desired. Simultaneous inhibition of metabolically and spatiotemporally related proteins/enzymes is a promising strategy for improving therapeutic interventions in cancer treatment. Herein, we report a poly-α-l-glutamate-based polymer inhibitor that simultaneously targets proximal transmembrane enzymes under hypoxia, namely, carbonic anhydrase IX (CAIX) and zinc-dependent metalloproteinases. A polymer incorporating two types of inhibitors more effectively inhibited the proliferation and migration of human breast cancer cells than a combination of two polymers functionalized exclusively with either inhibitor. Synergistic inhibition of cancer cells would occur owing to the hetero-multivalent interactions of the polymer with proximate enzymes on the cancer cell membrane. Our results highlight the potential of polymer-based cancer therapeutics.

Use of three-dimensional acceleration sensing to assess dairy cow gait and the effects of hoof trimming.

The aim of this study was to assess the use of three-dimensional acceleration sensing to describe the gait of dairy cows and the effect of hoof trimming. In Trial 1, a three-dimensional acceleration sensor was attached to the carpal region of a front leg of six Holstein cows who were then walked for 9 m. Results showed that measures of acceleration could clearly show the presence of steps, which were divided into an acceleration phase, which involved the hoof descending to and ascending from the ground, and a non-acceleration phase, which included the swing phase. Lissajous figures could succinctly depict the degree of acceleration for each cow and showed that one cow who was lame walked with greater vertical and lateral acceleration than the remaining cows. In Trial 2, 17 loose-housed Holstein cows were walked for 20 m during the month before, once during the month after and once 2 months after hoof trimming with the sensor attached at the end of their thoracic vertebrae to measure acceleration of the whole body. To relate the acceleration of the body to movements of the legs, image analysis was used to the range of vertical movement of four major skeletal joints, using visual markers attached to them. Hoof trimming significantly (P < 0.005) decreased both the range of vertical movement of the joints of front and hind legs and the variance of lateral and forward acceleration at the end of the thoracic vertebrae, suggesting improved gait pattern smoothness. Acceleration sensing was able to characterize variation among gait patterns of the cows and could reproduce the pattern shown by image analysis. This study demonstrates that three-dimensional acceleration sensing of either the carpal region of a front leg or the end of the thoracic vertebrae is useful to detect walking irregularities and evaluate the effectiveness of hoof trimming on walking ability of cattle.

Ability of heifers to discriminate between familiar herdmates and members of an unfamiliar group: preference test and operant conditioning test.

Using a preference test and operant conditioning in a Y-maze, this experiment examined the ability of heifers to discriminate between their own familiar herdmates and member(s) of an unfamiliar group. Sixteen Danish Friesian heifers, eight older animals (360.6 +/- 24.2 days of age) and eight younger ones (190.1 +/- 14.1 days of age) were used. Each age group was further divided into two experimental groups. Members of each of these groups were housed together in small pens before the experiments began. In experiment 1, each of the 16 animals was allowed to approach either a familiar or an unfamiliar individual in the Y-maze. The test was repeated 12 times, with a different unfamiliar subject for each test. In experiment 2, eight heifers were individually tested in a conditioning experiment to examine whether they could learn to discriminate between a group of their three herdmates and a group of three unfamiliar heifers. Test animals were rewarded when they chose their own group. In experiment 1, heifers did not show a preference between familiar and unfamiliar individuals. Interestingly the younger stimulus heifers but not the test animals showed an ability to discriminate between unfamiliar animals by vocalizing. In experiment 2, four of the eight test animals achieved the criterion for successful discrimination between the familiar and unfamiliar group (P < 0.003: binomial law). There was no age group difference in the ability to discriminate between familiar and unfamiliar animals. In conclusion, heifers did not show a preference toward familiar or unfamiliar individuals; but after conditioning, some heifers could learn to discriminate between familiar and unfamiliar groups.