Assessing Spns2-dependent S1P Transport as a Prospective Therapeutic Target.

S1P (sphingosine 1-phosphate) receptor modulator (SRM) drugs interfere with lymphocyte trafficking by downregulating lymphocyte S1P receptors. While the immunosuppressive activity of SRM drugs has proved useful in treating autoimmune diseases such as multiple sclerosis, that drug class is beset by on-target liabilities such as initial dose bradycardia. The S1P that binds to cell surface lymphocyte S1P receptors is provided by S1P transporters. Mice born deficient in one of these, spinster homolog 2 (Spns2), are lymphocytopenic and have low lymph S1P concentrations. Such observations suggest that inhibition of Spns2-mediated S1P transport might provide another therapeutically beneficial method to modulate immune cell positioning. We report here results using a novel S1P transport blocker (STB), SLF80821178, to investigate the consequences of S1P transport inhibition in rodents. We found that SLF80821178 is efficacious in a multiple sclerosis model but - unlike the SRM fingolimod - neither decreases heart rate nor compromises lung endothelial barrier function. Notably, although Spns2 null mice have a sensorineural hearing defect, mice treated chronically with SLF80821178 have normal hearing acuity. STBs such as SLF80821178 evoke a dose-dependent decrease in peripheral blood lymphocyte counts, which affords a reliable pharmacodynamic marker of target engagement. However, the maximal reduction in circulating lymphocyte counts in response to SLF80821178 is substantially less than the response to SRMs such as fingolimod (50% vs. 90%) due to a lesser effect on T lymphocyte sub-populations by SLF80821178. Finally, in contrast to results obtained with Spns2 deficient mice, lymph S1P concentrations were not significantly changed in response to administration of STBs at doses that evoke maximal lymphopenia, which indicates that current understanding of the mechanism of action of S1P transport inhibitors is incomplete.

The sensory mechanotransduction ion channel ASIC2 (acid sensitive ion channel 2) is regulated by neurotrophin availability.

Almost all sensory neurons of the dorsal root ganglia have a mechanosensitive receptive field in the periphery. We have shown that the sensitivity to mechanical stimuli of a subset of sensory neurons that are slowly adapting mechanoreceptors (SAM) is strongly dependent on the availability of brain-derived neurotrophic factor (BDNF). Here we have investigated whether the ASIC2 sodium channel, recently shown by us to be necessary for normal SAM sensitivity, might be regulated by BDNF and thus partially account for the down-regulation of SAM sensitivity seen in BDNF deficient mice. We show that the mRNA for ASIC2 channels is reduced in the DRG of BDNF deficient mice indicating that BDNF might maintain its expression in vivo. We also made short-term cultures of sensory neurons from adult BDNF deficient mice and used a specific antibody to detect the presence of ASIC2 channels in different classes of sensory neurons. We observed that the channel protein was dramatically down-regulated selectively in medium and large diameter neurons and this expression could be rescued in a dose and time dependent manner by addition of BDNF to the culture (10-100 ng/ml). Drugs that block new transcription or protein synthesis also prevented the rescue effects of BDNF. We observed that ASIC2 channels were down-regulated in sensory neurons taken from neurotrophin-4 and neurotrophin-3 deficient mice; these effects might be due to a selective loss of neurons that normally express large amounts of ASIC2 channels. In summary, our data identify the ASIC2 channel as a target of BDNF signaling in vivo and suggest that the functional down-regulation of sensory mechanotransduction in BDNF deficient mice is in part due to loss of ASIC2 expression.

Multichannel, direct-reading spectrograph attachment.

An instrument is described which provides a multichannel direct-reading capability for photoelectric spectrographic detection. The important feature of the instrument is the ability of the user to select at will any combination of channels for position adjustment and to move these while preserving the optical integrity of the enclosure. Photomultipliers are used and the results of performance tests are described.

The usefulness of minimal F-wave latency and sural/radial amplitude ratio in diabetic polyneuropathy.

The possibility of whether minimal F-wave latency and a simple ratio between the sural and superficial radial sensory response amplitudes may provide a useful electrodiagnostic test in diabetic patients was investigated in this report. To evaluate the diagnostic sensitivity of minimal F-wave latency, the Z-scores of the minimal F-wave latency, motor nerve conduction velocity (MCV), amplitude of compound muscle action potentials (CMAP), and distal latency (DL) of the median, ulnar, tibial, and peroneal nerve were compared in 37 diabetic patients. For the median, ulnar, and tibial nerves, the Z scores of the minimal F-wave latency were significantly larger than those of the MCV. In addition for all four motor nerves, the Z scores of the minimal F-wave latency were significantly larger than those for the CMAP amplitude. Furthermore, 19 subjects showing abnormal results in the standard sensory nerve conduction study had a significantly lower sural/radial amplitude ratio (SRAR), and 84% of them had an SRAR of less than 0.5. In conclusion, minimal F-wave latency and the ratio between the amplitudes of the sural and superficial radial sensory nerve action potential are sensitive measures for the detection of nerve pathology and should be considered in electrophysiologic studies of diabetic polyneuropathy.

Foot screening technique in a diabetic population.

Foot complications are a well known factor which contribute to the morbidity of diabetes and increases the chance of amputation. A total of 126 consecutive diabetic patients were evaluated by diabetic foot screening. Forty-one patients showed an impaired protective sense when tested with Semmes-Weinstein monofilament 5.07 (10 g), and 92% of them showed peripheral polyneuropathy in nerve conduction study (NCS). The mean vibration score of the Rydel-Seiffer graduated tuning fork in patients with peripheral polyneuropathy in nerve conduction (NCV) study was 5.38+/-2.0, which was significantly different from that of patients without polyneuropathy in NCS. Among the deformities identified on examination, callus, corn, and hallux valgus were the greatest. While checking the ankle/ brachial index (ABI), we also evaluated the integrity of vasculature in the lower extremities. After extensive evaluation, we classified the patients into eight groups (category 0,1,2,3,4A,4B,5,6). The result of this study suggested that the Semmes-Weinstein monofilament test, Rydel-Seiffer graduated tuning fork test, and checking the ankle/brachial index were simple techniques for evaluating pathologic change in the diabetic foot by office screening, and that this screening based on treatment-oriented classification helps to reduce pedal complications in a diabetic population.