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Despite great progress, thousands of neurodevelopmental disorder (NDD) risk genes remain to be discovered. We present a computational approach that accelerates NDD risk gene identification using machine learning. First, we demonstrate that models trained solely on single-cell RNA sequencing data can robustly predict genes implicated in autism spectrum disorder (ASD), developmental and epileptic encephalopathy (DEE), and developmental delay (DD). Notably, we find differences in gene expression patterns of genes with monoallelic and bi-allelic inheritance patterns in the developing human cortex. We then integrate expression data with 300 orthogonal features, including intolerance metrics, protein-protein interaction data, and others, in a semi-supervised machine learning framework (mantis-ml) to train inheritance-specific models for these disorders. The models have high predictive power (area under the receiver operator curves [AUCs]: 0.84-0.95), and the top-ranked genes were up to 2-fold (monoallelic models) and 6-fold (bi-allelic models) more enriched for high-confidence NDD risk genes compared to genic intolerance metrics alone. Additionally, genes ranking in the top decile were 45 to 180 times more likely to have literature support than those in the bottom decile. Collectively, this work provides robust NDD risk gene predictions that can complement large-scale gene discovery efforts and underscores the importance of considering inheritance in gene risk prediction., Competing Interests: Declaration of interests S.P. and D.V. are current employees and/or stockholders of AstraZeneca. R.S.D. and A.W.Z. have received consulting fees from AstraZeneca., (Copyright © 2025 American Society of Human Genetics. Published by Elsevier Inc. All rights reserved.)

𝜋-Conjugated polymers, including those based on acetylenic repeating units, are an exciting class of materials that offer narrow optical band gaps and tunable frontier orbital energies that lead to their use in organic electronics. This work expands the knowledge of structure-property relationships of acetylenic polymers through the synthesis and characterization of a series of Glaser-Hay-coupled model compounds and random copolymers comprised of BF 2 formazanate, fluorene, and/or bis(alkoxy)benzene units. The model compounds and copolymers synthesized exhibit redox activity associated with the reversible reduction of the BF 2 formazanate units and the irreversible reduction of the fluorene and bis(alkoxy)benzene units. The copolymers exhibit absorption profiles characteristic or intermediate of their respective models and homopolymers, leading to broad absorption of UV-vis light. The alkyne linkages of the model compounds and copolymers are reacted with [Co 2 (CO) 8 ] to convert the alkyne functional groups into cobalt carbonyl clusters. This transformation leads to blue-shifted absorption profiles due to a decrease in π-conjugation, demonstrating the ability to tune the properties of these materials through post-polymerization functionalization. The redox activity and broad absorption bands of the polymers reported make them excellent candidates for use in photovoltaics and other light-harvesting applications., (© 2024 The Author(s). Macromolecular Rapid Communications published by Wiley‐VCH GmbH.)

Since the Nobel prize winning discovery that polyacetylene could act as a semiconductor, there has been tremendous efforts dedicated to understanding and harnessing the unusual properties of π-conjugated polymers. Much of this research has focused on the preparation of oligoynes and polyynes with well-defined numbers of repeating alkyne units as models for carbyne. These studies are usually hampered by a structure-property relationship where the stability of the resulting materials decrease with the incorporation of additional alkyne units. Here, we describe a series of oligoynes, with up to 12 alkyne units, where electron-rich [Pt(PBu 3 ) 2 ] 2+ units are incorporated at the center of the oligoyne backbones which are capped by electron-poor BF 2 formazanate dyes. These compounds exhibit excellent stability and solubility, panchromatic absorption, and redox activity characteristic of their structural components. These traits facilitated thin-film studies of extended oligoyne materials, where it is shown that incorporating [Pt(PBu 3 ) 2 ] 2+ units leads to smoother films, decreased conductivity on the microscale, and increased conductivity on the nanoscale when compared to metal-free analogs. Remarkably, our oligoynes have superior conductivity compared to the ubiquitous poly(3-hexylthiophene) semiconductor., (© 2024 The Author(s). Chemistry - A European Journal published by Wiley-VCH GmbH.)

Spinocerebellar ataxia type 7 (SCA7) is an autosomal dominant neurological disorder caused by deleterious CAG repeat expansion in the coding region of the ataxin 7 gene (polyQ-ataxin-7). Infantile-onset SCA7 leads to severe clinical manifestation of respiratory distress, but the exact cause of respiratory impairment remains unclear. Using the infantile-SCA7 mouse model, the SCA7266Q/5Q mouse, we examined the impact of pathological polyQ-ataxin-7 on hypoglossal (XII) and phrenic motor units. We identified the transcript profile of the medulla and cervical spinal cord and investigated the XII and phrenic nerves and the neuromuscular junctions in the diaphragm and tongue. SCA7266Q/5Q astrocytes showed significant intranuclear inclusions of ataxin-7 in the XII and putative phrenic motor nuclei. Transcriptomic analysis revealed dysregulation of genes involved in amino acid and neurotransmitter transport and myelination. Additionally, SCA7266Q/5Q mice demonstrated blunted efferent output of the XII nerve and demyelination in both XII and phrenic nerves. Finally, there was an increased number of neuromuscular junction clusters with higher expression of synaptic markers in SCA7266Q/5Q mice compared with WT controls. These preclinical findings elucidate the underlying pathophysiology responsible for impaired glial cell function and death leading to dysphagia, aspiration, and respiratory failure in infantile SCA7.

The ongoing expansion of human genomic datasets propels therapeutic target identification; however, extracting gene-disease associations from gene annotations remains challenging. Here, we introduce Mantis-ML 2.0, a framework integrating AstraZeneca's Biological Insights Knowledge Graph and numerous tabular datasets, to assess gene-disease probabilities throughout the phenome. We use graph neural networks, capturing the graph's holistic structure, and train them on hundreds of balanced datasets via a robust semi-supervised learning framework to provide gene-disease probabilities across the human exome. Mantis-ML 2.0 incorporates natural language processing to automate disease-relevant feature selection for thousands of diseases. The enhanced models demonstrate a 6.9% average classification power boost, achieving a median receiver operating characteristic (ROC) area under curve (AUC) score of 0.90 across 5220 diseases from Human Phenotype Ontology, OpenTargets, and Genomics England. Notably, Mantis-ML 2.0 prioritizes associations from an independent UK Biobank phenome-wide association study (PheWAS), providing a stronger form of triaging and mitigating against underpowered PheWAS associations. Results are exposed through an interactive web resource.

Oligoynes and polyynes are 1D chains of conjugated sp-hybridized carbon atoms consisting of alternating single and triple bonds. Their stability rapidly decreases with increasing chain length beyond only a few repeating units. Design strategies, such as the use of bulky end-capping groups, allow for their characterization and isolation while not contributing significantly to their physical properties. In this study, we incorporate redox-active BF 2 formazanate dyes (BF 2 ) as end-caps to prepare symmetric (BF 2 -[C≡C] n -BF 2 ) and asymmetric (BF 2 -[C≡C] n -Si(iPr) 3 ) families of oligoynes containing up to 10 alkyne units. In doing so, we introduce stable oligoynes that possess a blend of optical and redox properties that cannot be achieved by either oligoynes or BF 2 formazanates individually (e.g., panchromatic absorption, multiple and tunable reversible redox waves). This approach is transferable to other functional end-caps to facilitate the preparation of π-conjugated materials with utility in the organic electronics arena., (© 2022 Wiley-VCH GmbH.)

π-Conjugated molecules with acceptor-donor-acceptor (A-D-A) electronic structures make up an important class of materials due to their tunable optoelectronic properties and applications in, for example, organic light-emitting diodes, nonlinear optical devices, and organic solar cells. The frontier molecular orbital energies, and thus band gaps, of these materials can be tuned by varying the donor and acceptor traits and π-electron counts of the structural components. Herein, we report the synthesis and characterization of a series of A-D-A compounds consisting of BF 2 formazanates as electron acceptors bridged by a variety of π-conjugated donors. The results, which are supported by density functional theory calculations, demonstrate rational control of optoelectronic properties and the ability to tune the corresponding band gaps. The narrowest band gaps (E g Opt = 1.38 eV and E g CV = 1.21 eV) were observed when BF 2 formazanates and benzodithiophene units were combined. This study provides significant insight into the band gap engineering of materials derived from BF 2 formazanates and will inform their future development as semiconductors for use in organic electronics.

Spinocerebellar ataxia type 7 (SCA7) is an autosomal-dominant neurodegenerative disorder caused by a CAG repeat expansion in the coding region of the ataxin-7 gene. Infantile-onset SCA7 patients display extremely large repeat expansions (>200 CAGs) and exhibit progressive ataxia, dysarthria, dysphagia and retinal degeneration. Severe hypotonia, aspiration pneumonia and respiratory failure often contribute to death in affected infants. To better understand the features of respiratory and upper airway dysfunction in SCA7, we examined breathing and putative phrenic and hypoglossal neuropathology in a knock-in mouse model of early-onset SCA7 carrying an expanded allele with 266 CAG repeats. Whole-body plethysmography was used to measure awake spontaneously breathing SCA7-266Q knock-in mice at baseline in normoxia and during a hypercapnic/hypoxic respiratory challenge at 4 and 8 weeks, before and after the onset of disease. Postmortem studies included quantification of putative phrenic and hypoglossal motor neurons and microglia, and analysis of ataxin-7 aggregation at end stage. SCA7-266Q mice had profound breathing deficits during a respiratory challenge, exhibiting reduced respiratory output and a greater percentage of time in apnea. Histologically, putative phrenic and hypoglossal motor neurons of SCA7 mice exhibited a reduction in number accompanied by increased microglial activation, indicating neurodegeneration and neuroinflammation. Furthermore, intranuclear ataxin-7 accumulation was observed in cells neighboring putative phrenic and hypoglossal motor neurons in SCA7 mice. These findings reveal the importance of phrenic and hypoglossal motor neuron pathology associated with respiratory failure and upper airway dysfunction, which are observed in infantile-onset SCA7 patients and likely contribute to their early death., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2021. Published by The Company of Biologists Ltd.)

Pompe disease is a lysosomal storage disease caused by mutations within the GAA gene, which encodes acid α-glucosidase (GAA)-an enzyme necessary for lysosomal glycogen degradation. A lack of GAA results in an accumulation of glycogen in cardiac and skeletal muscle, as well as in motor neurons. The only FDA approved treatment for Pompe disease-an enzyme replacement therapy (ERT)-increases survival of patients, but has unmasked previously unrecognized clinical manifestations of Pompe disease. These clinical signs and symptoms include tracheo-bronchomalacia, vascular aneurysms, and gastro-intestinal discomfort. Together, these previously unrecognized pathologies indicate that GAA-deficiency impacts smooth muscle in addition to skeletal and cardiac muscle. Thus, we sought to characterize smooth muscle pathology in the airway, vascular, gastrointestinal, and genitourinary in the Gaa -/- mouse model. Increased levels of glycogen were present in smooth muscle cells of the aorta, trachea, esophagus, stomach, and bladder of Gaa -/- mice, compared to wild type mice. In addition, there was an increased abundance of both lysosome membrane protein (LAMP1) and autophagosome membrane protein (LC3) indicating vacuolar accumulation in several tissues. Taken together, we show that GAA deficiency results in subsequent pathology in smooth muscle cells, which may lead to life-threatening complications if not properly treated.

The combination of π-conjugated organic compounds and Pt(ii)-acetylides is a powerful strategy for the production of functional optoelectronic materials. The presence of the heavy element, Pt, in these compounds enhances electronic delocalization generally resulting in low-energy absorption and emission maxima and often leads to intersystem crossing, resulting in phosphorescence. When boron complexes of N-donor ligands, such as boron dipyrromethenes (BODIPYs), are involved the molecular and polymeric materials produced have properties that are advantageous for their use as oxygen-sensors, in triplet-triplet annihilation, and as the functional components of photovoltaics. Based on these exciting results, we endeavored to thoroughly examine the effect of Pt(ii)-acetylide conjugation on the properties of BF2 formazanate dyes, which offer improved redox properties and red-shifted absorption and emission bands compared to many structurally related BODIPYs. The results showed that phosphine-supported Pt(ii)-acetylide incorporation enhanced electronic conjugation, rendering the electrochemical reduction of the BF2 formazanate dyes more difficult, while also red-shifting their absorption and emission maxima. Unlike similar BODIPYs, the presence of Pt(ii) did not facilitate phosphorescence, but rather quenched fluorescence. This study provides significant insights into structure-property relationships and guiding principles for the design of BF2 formazanate dyes, a rapidly emerging family of readily accessible optoelectronic materials.

Amyotrophic Lateral Sclerosis (ALS) is a devastating neurodegenerative disorder that results in death due to respiratory failure. Many genetic defects are associated with ALS; one such defect is a mutation in the gene encoding optineurin (OPTN). Using an optineurin null mouse (Optn -/- ), we sought to characterize the impact of optineurin deficiency on respiratory neurodegeneration. Respiratory function was assessed at 6 and 12 mo of age using whole body plethysmography at baseline during normoxia (FiO 2 : 0.21; N 2 balance) and during a respiratory challenge with hypoxia and hypercapnia (FiCO 2 : 0.07, FiO 2 : 0.10; N 2 balance). Histological analyses to assess motor neuron viability and respiratory nerve integrity were performed in the medulla, cervical spinal cord, hypoglossal nerve, and phrenic nerve. Minute ventilation, peak inspiratory flow, and peak expiratory flow are significantly reduced during a respiratory challenge in 6 mo Optn -/- mice. By 12 mo, tidal volume is also significantly reduced in Optn -/- mice. Furthermore, 12mo Optn -/- mice exhibit hypoglossal motor neuron loss, phrenic and hypoglossal dysmyelination, and accumulated mitochondria in the hypoglossal nerve axons. Overall, these data indicate that Optn -/- mice display neurodegenerative respiratory dysfunction and are a useful model to study the impact of novel therapies on respiratory function for optineurin-deficient ALS patients., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Amyotrophic lateral sclerosis (ALS) is a fatal disease characterized by degeneration of motor neurons and muscles, and death is usually a result of impaired respiratory function due to loss of motor neurons that control upper airway muscles and/or the diaphragm. Currently, no cure for ALS exists and treatments to date do not significantly improve respiratory or swallowing function. One cause of ALS is a mutation in the superoxide dismutase-1 ( SOD1 ) gene; thus, reducing expression of the mutated gene may slow the progression of the disease. Our group has been studying the SOD1 G93A transgenic mouse model of ALS that develops progressive respiratory deficits and dysphagia. We hypothesize that solely treating the tongue in SOD1 mice will preserve respiratory and swallowing function, and it will prolong survival. At 6 weeks of age, 11 SOD1 G93A mice (both sexes) received a single intralingual injection of gene therapy (AAVrh10-miR SOD1 ). Another 29 mice (both sexes) were divided into two control groups: (1) 12 SOD1 G93A mice that received a single intralingual vehicle injection (saline); and (2) 17 non-transgenic littermates. Starting at 13 weeks of age, plethysmography (respiratory parameters) at baseline and in response to hypoxia (11% O 2 ) + hypercapnia (7% CO 2 ) were recorded and videofluoroscopic swallow study testing were performed twice monthly until end-stage disease. Minute ventilation during hypoxia + hypercapnia and mean inspiratory flow at baseline were significantly reduced ( p  < 0.05) in vehicle-injected, but not AAVrh10-miR SOD1 -injected SOD1 G93A mice as compared with wild-type mice. In contrast, swallowing function was unchanged by AAVrh10-miR SOD1 treatment ( p  > 0.05). AAVrh10-miR SOD1 injections also significantly extended survival in females by ∼1 week. In conclusion, this study indicates that intralingual AAVrh10-miR SOD1 treatment preserved respiratory (but not swallowing) function potentially via increasing upper airway patency, and it is worthy of further exploration as a possible therapy to preserve respiratory capacity in ALS patients.

Duchenne muscular dystrophy (DMD) is a fatal neuromuscular disease caused by deleterious mutations in the DMD gene which encodes the dystrophin protein. Skeletal muscle weakness and eventual muscle degradation due to loss of dystrophin are well-documented pathological hallmarks of DMD. In contrast, the neuropathology of this disease remains understudied despite the emerging evidence of neurological abnormalities induced by dystrophin loss. Using quantitative morphological analysis of nerve sections, we characterize axonopathies in the phrenic and hypoglossal (XII) nerves of mdx mice. We observe dysfunction in these nerves - which innervate the diaphragm and genioglossus respectively - that we propose contributes to respiratory failure, the most common cause of death in DMD. These observations highlight the importance in the further characterization of the neuropathology of DMD. Additionally, these observations underscore the necessity in correcting both the nervous system pathology in addition to skeletal muscle deficits to ameliorate this disease.

Pompe disease is a glycogen storage disease caused by a deficiency in acid α-glucosidase (GAA), a hydrolase necessary for the degradation of lysosomal glycogen. This deficiency in GAA results in muscle and neuronal glycogen accumulation, which causes respiratory insufficiency. Pompe disease mouse models provide a means of assessing respiratory pathology and are important for pre-clinical studies of novel therapies that aim to treat respiratory dysfunction and improve quality of life. This review aims to compile and summarize existing manuscripts that characterize the respiratory phenotype of Pompe mouse models. Manuscripts included in this review were selected utilizing specific search terms and exclusion criteria. Analysis of these findings demonstrate that Pompe disease mouse models have respiratory physiological defects as well as pathologies in the diaphragm, tongue, higher-order respiratory control centers, phrenic and hypoglossal motor nuclei, phrenic and hypoglossal nerves, neuromuscular junctions, and airway smooth muscle. Overall, the culmination of these pathologies contributes to severe respiratory dysfunction, underscoring the importance of characterizing the respiratory phenotype while developing effective therapies for patients.

Boron difluoride (BF 2 ) formazanate dyes are contenders for molecular species that exhibit a large Stokes shift and bright red emission. Excitation of 3-cyanoformazanate complexes with 10 μs wide pulses of specific wavelengths resulted in strong luminescence at 663 nm at both room temperature in solution and at 77 K in a frozen solution. Analysis of the short-lived excitation spectrum from this luminescence shows that it arises from a vibronic manifold of a higher-lying excited state. This dark state relaxes to the emitting state over 10 μs. TD-DFT calculations of the two lowest-energy excited states show that the relaxed geometries are planar for S 1 but highly distorted in S 2 . The specific time- and wavelength-dependence of the excitation profile provides a unique optical encryption capability through the comparison of emission intensities between adjacent vibronic bands only accessible in the 0-12 μs time domain., (© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Amyotrophic lateral sclerosis (ALS) and spinocerebellar ataxia (SCA) are neurodegenerative disorders that result in progressive motor dysfunction and ultimately lead to respiratory failure. Rodent models of neurodegenerative disorders provide a means to study the respiratory motor unit pathology that results in respiratory failure. In addition, they are important for pre-clinical studies of novel therapies that improve breathing, quality of life, and survival. The goal of this review is to compare the respiratory phenotype of two neurodegenerative disorders that have different pathological origins, but similar physiological outcomes. Manuscripts reviewed were identified using specific search terms and exclusion criteria. We excluded manuscripts that investigated novel therapeutics and only included those manuscripts that describe the respiratory pathology. The ALS manuscripts describe pathology in respiratory physiology, the phrenic and hypoglossal motor units, respiratory neural control centers, and accessory respiratory muscles. The SCA rodent model manuscripts characterized pathology in overall respiratory function, phrenic motor units and hypoglossal motor neurons. Overall, a combination of pathology in the respiratory motor units and control centers contribute to devastating respiratory dysfunction.

The synthesis and characterization of polystannanes with "push" or "pull" moieties attached to the tin backbone are described. Precursor tetra aryl- (1, 2) stannanes were converted to mono- (3) and dichloro- (4, 5) stannanes by either sequential chlorination or by redistribution reactions with SnCl4. Compounds 4 and 5 were transformed to polymerisable tin dihydride monomers 6 and 7 using a large excess (10×) of NaBH4. Homopolymer 8 with electron donating aryl substituents (p-MeOC6H4-) was synthesized by dehydrogenative polymerization using Wilkinson's catalyst. Attempts to prepare the homopolymer 9 with electron withdrawing aryl substituents (p-CF3C6H4-) from the dehydrocoupling of 7 using similar conditions led only to the formation of low molecular weight oligomeric species. Two alternating polymers, 10 and 11, were synthesized by condensation polymerization of (n-Bu)2Sn(NEt2)2 with monomers 6 or 7. The first was a "push-push" alternating polymer, 10, comprised of a repeating unit consisting of two different electron donating groups (p-MeOC6H4-, n-Bu) at neighboring tin centres. The second was a "push-pull" alternating polymer, 11, bearing both an electron donating group (n-Bu) and a strongly electron withdrawing substituent (p-CF3C6H4-) at neighboring tin atoms. All small molecule stannanes and tin-containing polymers were characterized by NMR (1H, 13C, 119Sn, and where required 19F) spectroscopy, MS or EA. The absolute molecular weights of tin polymers (8, 10, 11) were determined by triple detection GPC and in the range of 1.07 × 104 to 1.95 × 104 Da. Rapid photodegradation of polymers was observed by UV-Vis spectroscopy, with a slower degradation observed for the "push-pull" polymer, 11, compared to the "push-push" polymer, 10.

The first example of a π-conjugated polymer incorporating boron difluoride (BF2) formazanates is introduced. The film-forming properties, controllable reduction chemistry, and low optical band gap (ca. 1.4 eV) of the polymer make it an excellent candidate for use as a light-harvesting n-type semiconductor in organic electronics. Comparison of the polymer to model compounds confirmed that its unique optoelectronic properties can be directly attributed to the presence of the BF2 formazanate repeat unit and that the [Pt(PBu3)2]2+ unit must also be present to achieve the narrow band gaps observed.

S-nitrosoglutathione (GSNO) is an endogenous nitric oxide (NO) carrier that plays a critical role in redox based NO signaling. Recent studies have reported that GSNO regulates the activities of STAT3 and NF-κB via S-nitrosylation dependent mechanisms. Since STAT3 and NF-κB are key transcription factors involved in tumor progression, chemoresistance, and metastasis of head and neck cancer, we investigated the effect of GSNO in cell culture and mouse xenograft models of head and neck squamous cell carcinoma (HNSCC). For the cell culture studies, three HNSCC cell lines were tested (SCC1, SCC14a and SCC22a). All three cell lines had constitutively activated (phosphorylated) STAT3 (Tyr(705)). GSNO treatment of these cell lines reversibly decreased the STAT3 phosphorylation in a concentration dependent manner. GSNO treatment also decreased the basal and cytokine-stimulated activation of NF-κB in SCC14a cells and reduced the basal low degree of nitrotyrosine by inhibition of inducible NO synthase (iNOS) expression. The reduced STAT3/NF-κB activity by GSNO treatment was correlated with the decreased cell proliferation and increased apoptosis of HNSCC cells. In HNSCC mouse xenograft model, the tumor growth was reduced by systemic treatment with GSNO and was further reduced when the treatment was combined with radiation and cisplatin. Accordingly, GSNO treatment also resulted in decreased levels of phosphorylated STAT3. In summary, these studies demonstrate that GSNO treatment blocks the NF-κB and STAT3 pathways which are responsible for cell survival, proliferation and that GSNO mediated mechanisms complement cispaltin and radiation therapy, and thus could potentiate the therapeutic effect in HNSCC., (Copyright © 2015. Published by Elsevier B.V.)

Sphingosine-1-phosphate receptor 1 (S1P1) mediated regulation of lymphocyte egress from lymphoid organs is recognized as the mechanism of FTY720 (Fingolimod, Gilenya) efficacy in relapsing-remitting forms of multiple sclerosis (RRMS). In this study we describe a novel S1P1 agonist AKP-11, next generation of S1P1 agonist, with immunomodulatory activities in cell culture model and for therapeutic efficacy against an animal model of MS, i.e. experimental autoimmune encephalomyelitis (EAE) but without the adverse effects observed with FTY720. Like FTY720, AKP-11 bound to S1P1 is internalized and activates intracellular AKT and ERKs cellular signaling pathways. In contrast to FTY720, AKP-11 mediated S1P1 downregulation is independent of sphingosine kinase activity indicating it to be a direct agonist of S1P1. The S1P1 loss and inhibition of lymphocyte egress by FTY720 leads to lymphopenia. In comparison with FTY720, oral administration of AKP-11 caused milder and reversible lymphopenia while providing a similar degree of therapeutic efficacy in the EAE animal model. Consistent with the observed reversible lymphopenia with AKP-11, the S1P1 recycled back to cell membrane in AKP-11 treated cells following its withdrawal, but not with withdrawal of FTY720. Accordingly, a smaller degree of ubiquitination and proteolysis of S1P1 was observed in AKP-11 treated cells as compared to FTY720. Consistent with previous observations, FTY720 treatment is associated with adverse effects of bradycardia and lung vascular leaks in rodents, whereas AKP-11 treatment had undetectable effects on bradycardia and reduced lung vascular leaks as compared to FTY720. Taken together, the data documents that AKP-11 treatment cause milder and reversible lymphopenia with milder adverse effects while maintaining therapeutic efficacy similar to that observed with FTY720, thus indicating therapeutic potential of AKP-11 for treatment of MS and related autoimmune disorders.

A study was conducted on the induction of buffalo sperm capacitation and acrosome reaction in the excised reproductive tract of hamsters at the estrogen- and progesterone-dominated stages of estrus. The percentages of the maximum capacitation and acrosome reaction were significatly (P < 0.01) higher for spermatozoa incubated in the uterus with oviducts of estrogen dominated hamsters compared with those incubated in BWW medium in a test tube (64.6%, 60.2%; 16.2%, 14.7%). Buffalo spermatozoa incubated in the uterus and oviducts of progesterone-dominated hamsters showed significantly (P < 0.01) lower capacitation and acrosome reaction rates than those incubated in the uterus and oviducts of estrogen-dominated hamsters (34.8%, 34.3%: 64.6%, 60.2%). The percentage of capacitation and acrosome reaction in spermatozoa were significantly (P < 0.01) more when incubated in the uterus plus oviducts than without the oviduct irrespective of whether the reproduct tract of hamster was estrogen- or progesterone-dominated. The time for the onset of maximum capacitation and acrosome reaction was reduced from 12 to 10 h when the spermatozoa were incubated in the hamster reproductive tract rather than in BWW medium in test tubes. The significance of the results in relation to hormonal regulation of sperm capaciation and acrosome reaction are also discussed.

A simple dual staining procedure for detecting the true acrosome reaction in dried smears of buffalo spermatozoa is described. Trypan blue is used first to differentiate live from dead spermatozoa and the dried smears which have been prepared are stained with Giemsa for acrosome evaluation. Four categories of spermatozoa were recognized: A) live, intact acrosome (acrosome pink, postnuclear cap clear); B) dead, intact acrosome (acrosome pink, postnuclear cap blue); C) live, detached acrosome (acrosome clear, postnuclear cap clear); and D) dead, detached acrosome (acrosome clear, postnuclear cap blue). The procedure is simple, rapid and convenient for assessing true acrosome reaction in buffalo spermatozoa. Simultaneous assessment of sperm viability and its acrosomal status in dried smears makes this procedure attractive because the true acrosome reaction can be studied thoroughly at a later state after the incubation period.