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The SARS-CoV-2 non-Spike (S) structural protein targets of nucleocapsid (N), membrane (M) and envelope (E), critical in the host cell interferon response and memory T-cell immunity, have been grossly overlooked since the inception of COVID vaccine development. To pursue a universal (pan-sarbecovirus) vaccine against ever-emergent future mutants, we explored booster immunogenicity of UB-612, a multitope-vaccine that contains S1-RBD-sFc protein and sequence-conserved rationally designed promiscuous Th and CTL epitope peptides on the Sarbecovirus N, M and S2 proteins. To a subpopulation of infection-free participants (aged 18-85 years) involved in a two-dose Phase-2 trial, a UB-612 booster (third dose) was administered 6-8 months after the second dose. The immunogenicity was evaluated at 14 days post-booster with overall safety monitored until the end of study. The booster induced high viral-neutralizing antibodies against live Wuhan WT (VNT50, 1,711) and Delta (VNT50, 1,282); and against pseudovirus WT (pVNT50, 11,167) vs. Omicron BA.1/BA.2/BA.5 variants (pVNT50, 2,314/1,890/854), respectively. The lower primary neutralizing antibodies in the elderly were uplifted upon boosting to approximately the same high level in young adults. UB-612 also induced potent, durable Th1-oriented (IFN-γ+-) responses (peak/pre-boost/post-boost SFU/106 PBMCs, 374/261/444) along with robust presence of cytotoxic CD8+ T cells (peak/pre-boost/post-boost CD107a+-Granzyme B+, 3.6%/1.8%/1.8%). Booster vaccination is safe and well tolerated without SAEs. By recognition against epitopes on Spike (S1-RBD and S2) and non-Spike (N and M) structure proteins, UB-612 provides potent, broad and long-lasting B-cell and T-cell memory immunity and offers a potential as a universal vaccine to fend off Omicrons and new VoCs.SIGNIFICANCE STATEMENTThe Omicron has swept the globe with a rapid succession of dominating sublineages from BA.1, BA.2, to the current BA.5 with increasing infectivity and antibody evasion. Concerningly, the non-Spike structure proteins that promote T-cell immunity are grossly overlooked in vaccine development. Looking beyond short-interval booster jabs and omicron-updated vaccines, a pragmatic approach to curbing ever-emergent new mutants would be “universal (pan-Sarbecovirus) vaccines” targeting conserved nonmutable epitopes on coronavirus. UB-612, a multitope-vaccine armed with Spike (S1-RBD and S2) and non-Spike targets (Nucleocapsid N and Membrane M), allows booster vaccination to elicit potent, broadly-recognizing, durable B- and T-cell memory immunity. Sequence-conserved epitope peptides were rationally-designed from S2, N and M proteins to synergistically enhance memory helper and cytotoxic T-cell immunity and B-cell immunity.

SARS-CoV-2 breakthrough infection occurs due to waning immunity time-to-vaccine, to which the globally-dominant, highly-contagious Delta variant is behind the scene. In the primary 2-dose and booster series of clinical Phase-1 trial, UB-612 vaccine, which contains S1-RBD and synthetic Th/CTL peptide pool for activation of humoral and T-cell immunity, induces substantial, prolonged viral-neutralizing antibodies that goes parallel with a long-lasting T-cell immunity; and a booster (3rd ) dose can prompt recall of memory immunity to induce profound, striking antibodies with the highest level of 50% viral-neutralizing GMT titers against live Delta variant reported for any vaccine. The unique design of S1-RBD only plus multitope T-cell peptides may have underpinned UB-612’s potent anti-Delta effect, while the other full S protein-based vaccines are affected additionally by mutations in the N-terminal domain sequence which contains additional neutralizing epitopes. UB-612, safe and well-tolerated, could be effective for boosting other vaccine platforms that have shown modest homologous boosting. [Funded by United Biomedical Inc., Asia; ClinicalTrials.gov ID: NCT04967742 and NCT04545749]

In the process of dielectronic recombination, an electron can be captured by a positive ion into doubly excited resonance states. These states can decay by autoionization or by radiative transitions which produce recombination lines. These lines are important to estimate the element abundances as well as the electron temperatures in planetary nebulae. The present paper reports a theoretical study by the R-matrix method for such processes of C2+ ions in planetary nebulae, which produce recombination lines ?5114 ?(4f'?3d'), ?4965 ?(3d'?3p'), ?4619 ?(4f'?3d') and ?8794 ?(3d'?3p'). Our study is based on three sets of target orbitals, through which we elucidate the electron correlation and static polarization effects in the dielectronic recombination processes. The calculated energy level positions are used to indicate the degrees of our calculation convergence; especially those based on the set of target orbitals including the electron correlation and static polarization effects agree well with the experimental data within about 4%. For such recombination processes, the recommended radiative transition rates, autoionization rates and net recombination rate coefficients are also provided. We anticipate that the accuracy of our recommended recombination rate coefficients for the whole four dielectronic lines is about 7% within our theoretical model.

Taxol is a highly effective anticancer agent that causes peripheral neuropathy as its major toxic side effect. The neuropathy is characterized by degeneration of sensory axons that may be severe enough to be dose limiting. Axonal degeneration involves the activation of the calcium-activated proteases calpains, and here we tested whether systemic inhibition of calpains with the peptide alpha-ketoamide calpain inhibitor AK295 can reduce the clinical and pathological effects of Taxol in a rodent model of Taxol neuropathy. In mice with Taxol neuropathy, AK295 reduced the degree of axonal degeneration in sensory nerve roots, and improved clinical measures of neuropathy, including behavioural and electrophysiological function. These findings were consistent for both 3- and 6-week models of neuropathy. In vitro, Taxol caused activation of both calpains and caspases in PC12 cells. AK295 inhibited the activation of calpains but did not interfere with the antimitotic effects of Taxol on microtubules, nor did it inhibit caspase-mediated cell death. These data implicate calpains in the pathogenesis of Taxol neuropathy, and demonstrate that AK295 can prevent axonal degeneration and clinical neuropathy in mice. In addition, AK295 did not interfere with the primary antineoplastic effects of Taxol on microtubules and cell death, suggesting that systemic calpain inhibition may be a good strategy for preventing neuropathy in patients being treated with Taxol.

The Wld s mouse is a spontaneous mutant that is characterized by the phenotype of delayed degeneration of transected nerves (slow Wallerian degeneration). Molecular genetic analysis identified a mutation in this animal that codes for a unique protein expressed in brain tissue of Wld s mice. We asked whether the Wld s phenotype, in addition to delaying axonal degeneration after axotomy, might provide neuroprotection against toxic neuropathy. In dorsal root ganglia (DRG) cultures, neurites from Wld s transiently exposed to vincristine not only resisted axonal degeneration but resumed growth after withdrawal of the toxin. Neurites from wild type mice died rapidly and did not recover. To prove that the identified mutation and its protein product are responsible for the Wld s phenotype, we used an adenoviral gene transfer system to deliver the Wld s to rat DRG neurons. Rat neurons expressing the Wld s protein were resistant to vincristine-induced axonal degeneration, confirming the functional significance of the identified gene mutation. These data provide evidence that the Wld s protein can be neuroprotective against vincristine neuropathy, and possibly other disorders characterized by axonal degeneration. In addition, delivery of this gene to wild type cells can transfer the Wld s phenotype, providing the possibility of gene therapy for peripheral neuropathy.

We recently showed that s.c. injections of a nonimmunosuppressant FK506 binding protein-12 (FKBP-12) ligand (V-10,367) accelerates nerve regeneration in the rat sciatic nerve crush model. Here we examined the oral efficacy of this compound for speeding nerve regeneration. Rats receiving V-10,367 (5, 15 or 50 mg/kg/day) by oral gavage all demonstrated an increase in nerve regeneration compared to vehicle-treated controls. Functional recovery was observed earliest and axonal calibers of regenerating axons in the soleus nerve were largest in the 15 mg/kg group, mean axonal areas being increased by 66% compared to controls. Orally active nonimmunosuppressant FKBP-12 ligands may be useful for the treatment of human peripheral nerve disorders.

The immunosuppressant drugs FK506 and cyclosporin A inhibit T-cell proliferation via a common mechanism: calcineurin inhibition following binding to their respective binding proteins, the peptidyl prolyl isomerases FKBP-12 and cyclophilin A. In contrast, FK506, but not cyclosporin A, accelerates nerve regeneration. In the present study, we show that the potent FKBP-12 inhibitor V-10,367, which lacks the structural components of FK506 required for calcineurin inhibition, increases neurite outgrowth in SH-SY5Y neuroblastoma cells and speeds nerve regeneration in the rat sciatic nerve crush model. In SH-SY5Y cells, V-10,367 increased the lengths of neurite processes in a concentration-dependent (between 1 and 10 nM) fashion over time (up to 168 h). Daily subcutaneous injections of V-10,367 accelerated the onset of clinical signs of functional recovery in the hind feet compared to vehicle-treated control animals. Interdigit distances (between the first and fifth digits) measured on foot prints obtained during walking showed an increase in toe spread in V-10,367-treated rats compared to vehicle-treated controls. Electron microscopy demonstrated larger regenerating axons distal to the crush site in the sciatic nerve from V-10,367-treated rats. Quantitation of axonal areas in the soleus nerve revealed a shift to larger axonal calibers in V-10,367-treated rats (400 or 200 mg/kg/day); mean axonal areas were increased by 52 and 59%, respectively, compared to vehicle-treated controls. FKBP-12 ligands lacking calcineurin inhibitory activity represent a new class of potential drugs for the treatment of human peripheral nerve disorders.

The effects of topical GM1 ganglioside on crushed rat sciatic nerve regeneration were studied in this presentation. Thirty-four rats, with 68 bilateral sciatic nerves, were divided into seven groups: one group of four rats for measurement of normal motor nerve conduction velocity (MNCV), three as controls, and another three in the GM1-treated groups. Sciatic nerves were exposed and crushed at a site 6 mm distal to the sciatic notch by the standard technique. In this manner, 3 mm wide crush injuries were created. Then 2.1 microliters of normal saline was injected into the crush site in the control groups and an equal volume of GM1 solution (containing 10 micrograms GM1) was injected into the GM1-treated groups. Electrophysiological, histological, and morphometric evaluations were carried out at 12, 28, and 56 days. A significantly higher muscle action potential (MAP) rate was found in the GM1-treated group (70%) vs. the controls (none) at 12 days (P < 0.005), and increased MNCV was found in the GM1-treated groups at both 28 and 56 days, especially at 56 days, when it was 39.59 +/- 9.23 m/sec vs. 31.42 +/- 4.07 m/sec in controls (P < 0.05). Morphometrically, there were more regenerated myelinated fibers (RMFs) at 12 days, and larger diameter of RMFs were observed at 12, 28, and 56 days in the GM1-treated groups.

The arterial plasma endothelin-1 concentration was substantially more elevated in 15 patients with rheumatic valvular disease and secondary pulmonary hypertension than in healthy volunteers (3.66 +/- 2.20 versus 1.17 +/- 0.38 pg/ml, mean +/- standard deviation; p < 0.01). The preoperative plasma endothelin-1 level was highly correlated with the pulmonary hemodynamics: pulmonary artery systolic pressure (r = 0.94, p < 0.001), pulmonary artery mean pressure (r = 0.86, p < 0.001), pulmonary capillary wedge pressure (r = 0.82, p < 0.001), and pulmonary vascular resistance (r = 0.63, p < 0.02). After valve replacement, the plasma endothelin-1 concentration declined substantially and the pulmonary hemodynamics improved markedly. Two weeks after the operation, the plasma endothelin-1 level in patients (1.26 +/- 0.45 pg/ml, mean +/- standard deviation) was not statistically different from that in the healthy volunteers. The plasma endothelin-1 concentration continuously increased during the course of cardiopulmonary bypass and peaked after cessation of bypass. The peak plasma endothelin-1 level (13.49 +/- 4.60 pg/ml, mean +/- standard deviation) positively correlated with the bypass time (r = 0.64, p < 0.02) and negatively correlated with the urine volume during bypass (r = -0.69, p < 0.01). We conclude that (1) increased plasma endothelin-1 might be implicated in the pathogenesis of secondary pulmonary hypertension caused by rheumatic valvular disease and (2) markedly elevated plasma endothelin-1 concentrations might be associated with the mechanism of cardiac or renal dysfunction after prolonged cardiopulmonary bypass.

The Ala499Val (CT) and Lys939Gln (AC) of the XPC gene are two potentially functional nonsynonymous polymorphisms, which affect the rate of DNA repair and might change XPC production and activity. This study aimed to explore the distribution of these two polymorphisms in the Chinese Han population and their relationship with male infertility.We genotyped the two polymorphisms of the XPC gene by the PCR-restriction fragment length polymorphism (PCR-RFLP) method in 318 infertile patients and 228 fertile male controls, detected the frequency of the alleles, and analyzed both the individual and the joint contribution of the two polymorphisms to male infertility.For the Ala499Val (CT) polymorphism, the frequencies of the CC, CT, and TT genotypes were significantly different in distribution between the patients and the controls (P = 0.020). Males with the TT genotype had a lower risk of male infertility than those with the CC genotype (adjusted OR = 0.49, 95% CI: 0.23-0.88), and even lower than those with both CC and CT genotypes (adjusted OR = 0.39, 95% CI: 0.22-0.71). The Lys939Gln (AC) polymorphism was not related with male infertility. The combined genotype analysis showed that the individuals with 1-4 risk alleles had a significantly higher risk of male infertility (adjusted OR = 2.75, 95% CI = 1.50-5.04) than those with 0 risk allele.The Ala499Val (CT) polymorphism of the XPC gene is correlated with male infertility and may be a potential genetic risk factor for male infertility in the Chinese Han population.

A flow controlling system for pulsed inhaled nitric oxide has been developed and tested, and here its features and initial animal experiments and clinical applications are described. The physical characteristic test indicates that the practical released dose of NO gas is very close to the theoretical flow of NO gas at variant pressures. Animal experiments demonstrate that inhaled NO gas concentration is lower than the concentration of theoretical inhalation, but the variance is not remarkable (p0.05). When sixteen cases with CHD and PH were chosen to inhale NO gas (15 ppm, 15 min) PAP and PVR of all cases were reduced after inhalation of NO gas from 617 +/-51.3 dyn x s x cm(-5), 54.4+/-13.1 mmHg to 417+/-36.9 dym x s x cm(-5), 33.8+/-12.3 mmHg (PVR, p0.01; PAP, p0.01) respectively. When gas inhalation was stopped, these values returned to their base lines after a short period of time. All these show that the pulsed inhaled NO flow controlling instrument in accordance with the requirements of the designing, can be widely used in clinical diagnoses and treatments and will be a new tool offered for the treatments of the patients with PH.

Neurotrophic activity of neuroimmunophilin ligands (FK506 and its nonimmunosuppressant derivatives) has been assumed to be mediated by the FK506-binding protein-12 (FKBP-12). We recently showed that activity is retained in hippocampal neurons from FKBP-12 knockout mice, indicating that binding to FKBP-12 is not necessary. Here we show that three nonimmunosuppressant FK506 derivatives (V-13,450, V-13,629, and V-13,670) that do not bind FKBP-12 (>12.5 mM affinity) are equipotent to FKBP-12 ligands (FK506, V-10,367, and V-13,449) for increasing neurite elongation in SH-SY5Y cells. One non-FKBP-12 ligand (V-13,670) is also shown to accelerate functional recovery and nerve regeneration in the rat sciatic nerve crush model. Surprisingly, it exhibited an unusual dose-response effect upon oral administration, showing a novel bimodal dose-response for behavioral functional recovery and myelination, but not for axonal size, suggesting both Schwann cell and neuronal targets. Orally active non-FKBP-12 neuroimmunophilin ligands may be useful for the treatment of human neurological disorders without any potential side effects resulting from FKBP-12 binding.

The conduction of nonoriented, hazy polypropylene films has been investigated between 8 degrees C and 123 degrees C at electrical stresses of up to 20 kV DC using a ramping speed of 200 V/s and an inert atmosphere. The film thickness ranged from 70 to 80 mu m. The recorded current traces indicate unusual behavior in the vicinity of 113 degrees C. High-resolution differential scanning calorimetry also suggested a departure from linear behavior at this temperature. An attempt is made to explain this behavior in terms of structural changes that lead to the development of a negative resistance. >

For amorphous nonpolar polymers, a quantitative relation between dielectric constant and free volume was extended from the authors' (1990) previous model for liquids. An expression of ionic conductivity in terms of free volume was developed based on the Nernst-Einstein equation. Furthermore, the correlation between ionic conductivity and dielectric constant was derived. Experimental results on polystyrene agree well with the theoretical models over a wide range of temperature (-180 degrees C to +140 degrees C). >

A formula for the permittivity expansion of the Claussius-Mossotti function is derived using a virial formalism. The liquids considered consist of nonpolar, axial molecules possessing permanent quadrupole moments and anisotropic polarizability. The second and third permittivity virial coefficients can be evaluated on the basis of two and three body interactions; the intermolecular potential used is of the Lennard-Jones form. The results are in satisfactory agreement with experimental data on CCl/sub 4/, CH/sub 4/, C/sub 2/H/sub 6/, C/sub 3/H/sub 8/, and n-alkanes, (C/sub 4/ to C/sub 10/). >

A relation between relative dielectric constant and fractional free volume (FFV) of nonpolar condensed matter was previously developed by the authors using the Clausius-Mosotti equation together with free volume theory. A comparison of this relation with a large body of experimental data collected from the literature is given. It is concluded that this relation coincides quite satisfactorily with the characteristics of n-alkanes (from C/sub l/ to C/sub l2/) over a wide range of temperatures. >

The voltage-current characteristics of dodecyl benzene (DDB), transformer oil, mineral oil, and silicone oil have been investigated at different temperatures under ramp voltage (up until breakdown occurred) at a ramping speed of 100 V/s. Mercury was used as an electrode of sphere (Al or Cu)-plate (Hg) electrode systems. The work function of Hg derived from the electron emission (or charge injection) mechanisms was on the order of a few electronvolts, which was much larger than those of Al and Cu in the same case. This was attributed to the existence of few surface states because of Hg's smooth surface caused by the liquid surface tension. >

The WldS mouse is a unique mutant strain that demonstrates the remarkable phenotype of prolonged survival of transected axons ("slow Wallerian degeneration"). In these studies, we tested whether this neuroprotective phenotype extends to axonal degeneration seen in a progressive peripheral neuropathy. WldS and wild-type mice were intoxicated with the cancer chemotherapeutic agent paclitaxel (Taxol). The severity of the resultant sensory neuropathy was compared with behavioral, physiological, and pathological measures. WldS mice were resistant to paclitaxel neuropathy by all measures, and the resistance was because of protection against axonal degeneration. These studies demonstrate for the first time that the WldS mouse is more than a slow Wallerian degeneration phenotype, emphasizing the mechanistic link between Wallerian degeneration and peripheral neuropathy. Understanding how this mutant gene confers protection against axonal degeneration will provide important clues toward prevention of axonal degeneration in several human neurological disorders.

The authors examined the ability of FK506 to accelerate axonal regeneration of rat spinal cord axons in a peripheral nerve (PN) graft. Predegenerated autografts were produced by transecting the left tibial nerve 1 week prior to spinal cord implantation into the lumbar (L-3-L-4) spinal cord. Rats were given daily injections of either FK506 (5 mg/kg, subcutaneous) or vehicle for 21 days. The PN grafts from FK506-treated rats contained larger sized regenerating axons compared with vehicle-treated controls, and mean axonal areas increased by 25% at 7.5 mm along the PN graft. Fluoro-Gold retrograde labeling confirmed that the regenerating axons originated from the central nervous system. Unexpectedly, the majority (>50%) of neurons in the red nucleus were retrogradely labeled in the FK506-treated animals only. The results indicate that FK506 not only accelerates the elongation of spinal cord axons but also promotes regeneration of rubrospinal neurons.

We examined whether the nerve regenerative property of FK506 exhibits a 'window-of-opportunity' corresponding to the time of injury for maximal efficacy in the sciatic nerve crush model. FK506 (5 mg/kg, s.c.) was administered over the 18-day period of study according to three dosage regiments: delayed (days 9-17), discontinuous (days 0-8) and continuous (days 0-17) administrations. Quantitation of axonal calibers and the extend of myelination in the soleus nerve at 18 days demonstrated that both delayed and discontinuous administrations were equally effective, arguing against a 'window-of-opportunity' for FK506 nerve regenerative effect. However, both protocols were less effective than continuous administration indicating that the compound needs to be given during the entire regenerative period to elicit maximal efficacy.

The effects of nerve growth factor (NGF) on crushed sciatic nerve regeneration were studied in 30 rats, with 60 bilateral nerves. The nerves were crushed at a site 6 mm distal to the sciatic notch by the standard technique and 3 mm wide crush injuries were created. Then 2.1 microliters of normal saline in the control groups and an equal volume of NGF solution (containing 1 microgram of NGF) in the NGF-treated groups was injected into the crush sites and followed for 12, 28, and 56 days, respectively. At the end of the observation, electrophysiological evaluation was carried out; then samples 10 mm distal to the crush site were removed and prepared for histological and morphometric studies. Evoked muscle action potential (MAP) was recorded in 50% of the NGF-treated group at 12 days but not in the control group; the difference was statistically significant (P < 0.05). The motor nerve conduction velocity (MNCV) was increased in NGF-treated groups compared with control groups at 28 and 56 days (P < 0.05). Morphometrically, significantly more regenerated myelinated fibers (RMFs) were seen at 12 days, and larger diameter RMFs were found at 12, 28, and 56 days in NGF-treated groups than in control groups. These results indicate that topically applied NGF stimulates nerve regeneration and promotes function recovery in crushed rat sciatic nerves.

The implantation of a microvascular polyester prosthesis (MPP) (internal diameter 1 mm, length 10 mm) in the carotid arteries was studied in 48 Sprague-Dawley rats. The left carotid artery was resected to 7 mm in length and replaced with MPP by means of sleeve anastomoses. Postoperatively the patency was monitored with Doppler ultrasound at 24 and 72 hr, and at 1, 2, 3, 4, 8, and 12 weeks and was confirmed either by direct inspection intraoperatively or by angiography at the time of removal. At each monitoring time, all the throm-bosed and some selected patent grafts were removed and prepared for scanning electron microscopy (SEM) study. The early patency rate was 100% within 24 hr, and the cumulative long-term patency rate was 77.1% at 12 weeks. Rapid and complete endothelial lining (EL) restoration was demonstrated 3 weeks after operation, which provided a smooth and nonthrombogenic surface, and contributed to the long-term patency. Ten of the 11 failed grafts occurred within 3 weeks, that is, before EL, due to thrombosis. These observations indicate that MPP can be successfully used as arterial conduits in rats, and an enhanced patency rate may be expected with the use of anticoagulants or endothelial cell seeding for prevention of early thrombosis. © 1994 Wiley-Liss, Inc. Foreign Text Ignored. © 1994 Wiley-Liss, Inc.