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Extensive monitoring of particulate matter (PM) smaller than 2.5 µm, i.e., PM2.5, is critical for understanding changes in local air quality due to policy measures. With the emergence of low-cost air quality sensor networks, high spatiotemporal measurements of air quality are now possible. However, the sensitivity, noise, and accuracy of field data from such networks are not fully understood. In this study, we use spectral analysis of a 2-year data record of PM2.5 from both the Environmental Protection Agency (EPA) and PurpleAir (PA), a low-cost sensor network, to identify the contributions of individual periodic sources to local air quality in Chicago. We find that sources with time periods of 4, 8, 12, and 24 h have significant but varying relative contributions to the data for both networks. Further analysis reveals that the 8 and 12 h sources are traffic-related and photochemistry-driven, respectively, and that the contributions of both these sources are significantly lower in the PA data than in the EPA data. The presence of distinct peaks in the power spectrum analysis highlights recurring patterns in the air quality data; however, the underlying factors contributing to these peaks require further investigation and validation. We also use a correction model that accounts for the contribution of relative humidity and temperature, and we observe that the PA temporal components can be made to match those of the EPA over the medium and long term but not over the short term. Thus, standard approaches to improve the accuracy of low-cost sensor network data will not result in unbiased measurements. The strong source dependence of low-cost sensor network measurements demands exceptional care in the analysis of ambient data from these networks, particularly when used to evaluate and drive air quality policies.

Abstract Members of the highly polymorphic Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) family expressed on the surface of infected erythrocytes (IEs) are important virulence factors, which mediate vascular adhesion of IEs via endothelial host receptors and are targets of naturally acquired immunity. The PfEMP1 family can be divided into clinically relevant subgroups, of which some bind intercellular adhesion molecule 1 (ICAM-1). While the acquisition of IgG specific for ICAM-1-binding DBLβ domains is known to differ between PfEMP1 groups, its ability to induce antibody-dependent cellular phagocytosis (ADCP) is unclear. We therefore measured plasma levels of DBLβ-specific IgG, the ability of such IgG to inhibit PfEMP1-binding to ICAM-1, and its ability to opsonize IEs for ADCP, using plasma from Beninese children with severe (SM) or uncomplicated malaria (UM). IgG specific for DBLβ from group A and B ICAM-1-binding PfEMP1 were dominated by IgG1 and IgG3, and were similar in SM and UM. However, levels of plasma IgG inhibiting ICAM-1-binding of group A DBLβ of PFD1235w was significantly higher in children with UM than SM, and acute UM plasma induced a higher ADCP response than acute SM plasma.

Abstract Killer whales (Orcinus orca) are top predators throughout the world’s oceans. In the North Pacific, the species is divided into three ecotypes—resident (fish-eating), transient (mammal-eating), and offshore (largely shark-eating)—that are genetically and acoustically distinct and have unique roles in the marine ecosystem. In this study, we examined the year-round distribution of killer whales in the northern Gulf of Alaska from 2016 to 2020 using passive acoustic monitoring. We further described the daily acoustic residency patterns of three killer whale populations (southern Alaska residents, Gulf of Alaska transients, and AT1 transients) for one year of these data. Highest year-round acoustic presence occurred in Montague Strait, with strong seasonal patterns in Hinchinbrook Entrance and Resurrection Bay. Daily acoustic residency times for the southern Alaska residents paralleled seasonal distribution patterns. The majority of Gulf of Alaska transient detections occurred in Hinchinbrook Entrance in spring. The depleted AT1 transient killer whale population was most often identified in Montague Strait. Passive acoustic monitoring revealed that both resident and transient killer whales used these areas much more extensively than previously known and provided novel insights into high use locations and times for each population. These results may be driven by seasonal foraging opportunities and social factors and have management implications for this species.

It is of international priority to develop a vaccine against sexually transmitted Chlamydia trachomatis infections to combat the continued global spread of the infection. The optimal immunization strategy still remains to be fully elucidated. The aim of this study was to evaluate immunization strategies in a nonhuman primate (NHP) model. Cynomolgus macaques (Macaqua fascicularis) were immunized following different multi-component prime-boost immunization-schedules and subsequently challenged with C. trachomatis SvD in the lower genital tract. The immunization antigens included the recombinant protein antigen CTH522 adjuvanted with CAF01 or aluminium hydroxide, MOMP DNA antigen and MOMP vector antigens (HuAd5 MOMP and MVA MOMP). All antigen constructs were highly immunogenic raising significant systemic C. trachomatis-specific IgG responses. In particularly the CTH522 protein vaccinated groups raised a fast and strong pecificsIgG in serum. The mapping of specific B cell epitopes within the MOMP showed that all vaccinated groups, recognized epitopes near or within the variable domains (VD) of MOMP, with a consistent VD4 response in all animals. Furthermore, serum from all vaccinated groups were able to in vitro neutralize both SvD, SvE and SvF. Antibody responses were reflected on the vaginal and ocular mucosa, which showed detectable levels of IgG. Vaccines also induced C. trachomatis-specific cell mediated responses, as shown by in vitro stimulation and intracellular cytokine staining of peripheral blood mononuclear cells (PBMCs). In general, the protein (CTH522) vaccinated groups established a multifunctional CD4 T cell response, whereas the DNA and Vector vaccinated groups also established a CD8 T cells response. Following vaginal challenge with C. trachomatis SvD, several of the vaccinated groups showed accelerated clearance of the infection, but especially the DNA group, boosted with CAF01 adjuvanted CTH522 to achieve a balanced CD4/CD8 T cell response combined with an IgG response, showed accelerated clearance of the infection.

Per- and polyfluoroalkyl substances (PFAS) are a broad class of synthetic chemicals; some are present in most humans in developed countries. Several studies have shown associations between certain PFAS, such as perfluorooctanoic acid (PFOA) and perfluorooctane sulfonic acid (PFOS), and reduced antibody concentration after vaccination against diseases such as Tetanus. Recent studies have reported associations between COVID-19 occurrence and exposure to certain types of PFAS. However, studies of antibody concentration after COVID-19 vaccination in relation to PFAS serum concentrations have not been reported. We examined COVID-19 antibody responses to vaccines and PFAS serum concentrations among employees and retirees from two 3M facilities, one of which historically manufactured PFOS, PFOA, and perfluorohexane sulfonic acid (PFHxS).Participants completed enrollment and follow-up study visits in the Spring of 2021, when vaccines were widely available. In total 415 participants with 757 observations were included in repeated measures analyses. Log-transformed concentrations of anti-spike IgG and neutralizing antibodies were modeled in relation to concentration of PFAS at enrollment after adjusting for antigenic stimulus group (9 groups determined by COVID-19 history and number and type of vaccination) and other variables. The fully adjusted IgG concentration was 3.45 percent lower (95% CI −7.03, 0.26) per 14.5 ng/mL (interquartile range) increase in PFOS; results for neutralizing antibody and PFOS were similar. For PFOA, PFHxS, and perfluorononanoic acid (PFNA), the results were comparable to those for PFOS, though of smaller magnitude. In our study data, the fully adjusted coefficients relating concentration of vaccine-induced antibodies to COVID-19 and interquartile range difference in serum concentration of PFOS, PFOA, PFHxS, and PFNA were inverse but small with confidence intervals that included zero. Our analysis showed that the coefficient for the four PFAS examined in detail was considerably affected by adjustment for antigenic stimulus group.

Abstract Prolactin (PRL) hormone functions as a pleiotropic cytokine with a protective role in the retina. We recently identified by transcriptome profiling that PRL is one of the most highly upregulated mRNAs in the retinas of mutant rcd1 (PDE6B) and xlpra2 (RPGR) dogs at advanced stages of photoreceptor disease. In the present study, we have identified the expression of a short PRL isoform that lacks exon 1 in canine retinas and analyzed the time-course of expression and localization of this isoform in the retinas of these two models. Using laser capture microdissection to isolate RNA from each of the retinal cellular layers, we found by qPCR that this short PRL isoform is expressed in photoreceptors of degenerating retinas. We confirmed by in situ hybridization that its expression is localized to the outer nuclear layer and begins shortly after the onset of disease at the time of peak photoreceptor cell death in both models. PRL protein was also detected only in mutant dog retinas. Our results call for further investigations into the role of this novel PRL isoform in retinal degeneration.

Purpose: Myeloproliferative neoplasms (MPNs) have been associated with increased systemic levels of vascular endothelial growth factor (VEGF). This study investigated an association between systemic busulfan for treatment of MPN and the requirement for intravitreal anti-VEGF injections for treatment of retinal pathology. Methods: Retrospective chart review of patients receiving systemic busulfan for myeloproliferative neoplasm and intravitreal anti-VEGF injections for macular and retinal vascular diseases from 2007 to 2021. Results: Of seven patients receiving oral busulfan for a hematological neoplasm and having concomitant retinal pathology requiring intravitreal anti-VEGF, all were white females >60 years old with MPN and exudative age-related macular degeneration. Of these, two patients had a reduced anti-VEGF requirement while on systemic busulfan, two took busulfan for fewer than 5 months, one developed retinal pathology over one year after stopping busulfan, one developed new retinal pathology while taking busulfan, and one had limited follow-up. Of the two patients with reduced anti-VEGF requirement while taking systemic busulfan, both had JAK2 V617F mutated MPN, and subsequent busulfan discontinuation was associated with an increased requirement for intravitreal anti-VEGF injections. Conclusions and importance: Systemic busulfan for treatment of MPN was associated with a reduced requirement for intravitreal anti-VEGF injections for retinal vascular disease in two patients. This association could be a result of inhibition of proliferative angiogenesis or reduced systemic VEGF levels with effective systemic treatment for MPN. Further study is required to confirm this association and determine whether this relationship is specific to busulfan or extends to other systemic medications used to treat MPN.

The Dingsishan Site, located in Nanning City, Guangxi Zhuang Autonomous Region, is one of the most important Neolithic archaeological sites in the Lingnan region of China’s southeastern seaboard. Plant microfossil remains recovered from excavated artifacts and human teeth suggest that the site’s ancient inhabitants practiced a subsistence system based on foraging. Wild plant food resources dominated their vegetal diet. Starch granules extracted from residue samples represent various taxa, including plant roots and tubers, aquatic plant fruits, beans, and wild cereals, defining the primary vegetal diet of Dingsishan’s Neolithic occupants. In addition, residue samples from shell artifacts yielded starch granules and phytolith remains, providing significant clues as to the function of these tools. We also identified millet starch granules from Dingsishan Cultural Phases 3-4 (ca. 7,000–6000 BP), albeit in extremely low quantities. Holocene paleoecological conditions in the Lingnan area provided diverse and plentiful wild food resources, allowing the relatively late emergence of agriculture. Our study suggests that Middle Neolithic human groups in the Yong River drainage consumed various plants, and their subsistence pattern was relatively stable through the four Cultural Phases identified at Dingsishan. Our conclusions enhance understanding the diversity of plant food resources exploited by foraging societies and interpretations of differences in patterns of agricultural origins in different ecological regions of China.

Purpose: To present a case of subretinal abscess associated with pneumonia and meningitis caused by Cryptococcus gattii in an immunocompetent host. Observations: A 37-year-old man presented with sub-acute painless unilateral vision loss and a white submacular elevation. Systemic evaluation revealed a lung lesion and cerebrospinal fluid evidence of Cryptococcus gattii infection. Conclusions and importance: While Crypococcus neoformans has been well described as a cause of chorioretinitis in immunocompetent and immunocompromised hosts, this report demonstrates that Cryptocuccus gattii is a related uncommon pathogen to be considered in similar presentations. Submacular surgical debridement may be challenging and OCT imaging may be helpful to detect full-thickness retinal necrosis.

Throughout the arid lands of Africa and Eurasia, camelids facilitated the expansion of human populations into areas that would not likely have been habitable without the transportation abilities of this animal along with the organic resources it provides, including dung, meat, milk, leather, wool, and bones. The two-humped, Bactrian, species of Camelus, C. ferus in its wild state and C. bactrianus when domesticated, is much more poorly known in prehistoric archaeological contexts than its single-humped congeneric, C. dromedarius. Our research uses a convergence of evidence approach to analyze reports and remains of Plio-Pleistocene camelids in Central and Northern Asia and trace the latest-known fossil Bactrian relative, Camelus knoblochi, that seems to have survived in the Gobi Desert until the Last Glacial Maximum (ca. 26.5–19 ka). Rock art depictions, some of which may be of Pleistocene age, record the complexity of nascent human-camel interactions and provide the impetus for further archaeological studies of both the origins of C. bactrianus and its increasingly complex relationships with the highly mobile prehistoric peoples of Central and Northern Asia.

Background Millions of workers are potentially exposed to respirable crystalline silica (RCS) which has been associated with several diseases. We updated the mortality experience of a cohort of 2,650 mine and mill workers at four manufacturing facilities to assess cause-specific mortality risks associated with estimated cumulative RCS exposure. Methods Study eligibility was defined as any employee who had ≥1 year of service by 2000, with work history experience available from 1945 through 2004. Vital status and cause of death were ascertained from 1945 through 2015. RCS exposure was estimated across plant-, department-, job-, and time-dependent categories using historic industrial hygiene sampling data and professional judgment. Associations between cumulative RCS (mg/m3-years) and cause-specific mortality were examined using Cox proportional hazard regression models. Results In the exposure-response analysis defined on quartiles of cumulative RCS exposure, no increasing trend (ptrend = 0.37) in lung cancer mortality (n = 116 deaths) was observed (Hazard ratio (HR) = 1.00 (referent), 1.20, 1.85, 0.92). Mortality risk for non-malignant respiratory disease was increased across quartiles (HR = 1.00, 1.35, 1.89, 1.70; ptrend = 0.15), based on 83 deaths. Non-malignant renal disease mortality was increased across quartiles (HR = 1.00, 6.64, 3.79, 3.29; ptrend = 0.11), based on 26 deaths. Conclusions After nearly seven decades of follow-up, the exposure-response analyses showed no evidence of a positive trend for lung cancer, and limited evidence of a trend for non-malignant respiratory disease, and non-malignant renal disease mortality as a result of cumulative RCS exposure in this occupational cohort.

The Liangwangcheng site, located in Pizhou County, Xuzhou City, northern Jiangsu Province, is one of the most important Neolithic Dawenkou Culture archeological sites in the Haidai area of China’s eastern seaboard. In recent years, archaeobotanical studies in the Haidai area, mainly focusing on Shandong Province, have yielded fruitful results, while relatively few such studies have been undertaken in northern Jiangsu Province. Here, we report the results of dental residue analysis conducted on 31 individual human skulls unearthed from the Late Dawenkou Culture Liangwangcheng site. The starch granules extracted from these residue samples indicate that foxtail and broomcorn millet, rice, roots and tubers, and legumes comprised the vegetal diet of Liangwangcheng’s occupants. Evidence suggests that mixed rice–millet agriculture played a definite role, with the coexistence of gathering as an economic element. According to archaeobotanical evidence from surrounding cotemporaneous sites, the Late Neolithic human groups that lived in the lower Huang-Huai River drainage shared similar subsistence patterns. Our results provide new evidence for a more comprehensive understanding of plant resource utilization and agricultural development in northern Jiangsu during the Dawenkou period.

Abstract Background Duplications of 15q11.2-q13.1 (Dup15q syndrome), including the paternally imprinted gene UBE3A and three nonimprinted gamma-aminobutyric acid type-A (GABAA) receptor genes, are highly penetrant for neurodevelopmental disorders such as autism spectrum disorder (ASD). To guide targeted treatments of Dup15q syndrome and other forms of ASD, biomarkers are needed that reflect molecular mechanisms of pathology. We recently described a beta EEG phenotype of Dup15q syndrome, but it remains unknown which specific genes drive this phenotype. Methods To test the hypothesis that UBE3A overexpression is not necessary for the beta EEG phenotype, we compared EEG from a reference cohort of children with Dup15q syndrome (n = 27) to (1) the pharmacological effects of the GABAA modulator midazolam (n = 12) on EEG from healthy adults, (2) EEG from typically developing (TD) children (n = 14), and (3) EEG from two children with duplications of paternal 15q (i.e., the UBE3A-silenced allele). Results Peak beta power was significantly increased in the reference cohort relative to TD controls. Midazolam administration recapitulated the beta EEG phenotype in healthy adults with a similar peak frequency in central channels (f = 23.0 Hz) as Dup15q syndrome (f = 23.1 Hz). Both paternal Dup15q syndrome cases displayed beta power comparable to the reference cohort. Conclusions Our results suggest a critical role for GABAergic transmission in the Dup15q syndrome beta EEG phenotype, which cannot be explained by UBE3A dysfunction alone. If this mechanism is confirmed, the phenotype may be used as a marker of GABAergic pathology in clinical trials for Dup15q syndrome.

Abstract Background Age-related macular degeneration (AMD) is a degenerative disorder of the central retina and the foremost cause of blindness. The retinal pigment epithelium (RPE) is a primary site of disease pathogenesis. The genetic basis of AMD is relatively well understood; however, this knowledge is yet to yield a treatment for the most prevalent non-neovascular disease forms. Therefore, tissue-specific epigenetic mechanisms of gene regulation are of considerable interest in AMD. We aimed to identify differentially methylated genes associated with AMD in the RPE and differentiate local DNA methylation aberrations from global DNA methylation changes, as local DNA methylation changes may be more amenable to therapeutic manipulation. Methods Epigenome-wide association study and targeted gene expression profiling were carried out in RPE cells from eyes of human donors. We performed genome-wide DNA methylation profiling (Illumina 450k BeadChip array) on RPE cells from 44 human donor eyes (25 AMD and 19 normal controls). We validated the findings using bisulfite pyrosequencing in 55 RPE samples (30 AMD and 25 normal controls) including technical (n = 38) and independent replicate samples (n = 17). Long interspersed nucleotide element 1 (LINE-1) analysis was then applied to assess global DNA methylation changes in the RPE. RT-qPCR on independent donor RPE samples was performed to assess gene expression changes. Results Genome-wide DNA methylation profiling identified differential methylation of multiple loci including the SKI proto-oncogene (SKI) (p = 1.18 × 10−9), general transcription factor IIH subunit H4 (GTF2H4) (p = 7.03 × 10−7), and Tenascin X (TNXB) (p = 6.30 × 10−6) genes in AMD. Bisulfite pyrosequencing validated the differentially methylated locus cg18934822 in SKI, and cg22508626 within GTF2H4, and excluded global DNA methylation changes in the RPE in AMD. We further demonstrated the differential expression of SKI, GTF2H4, and TNXB in the RPE of independent AMD donors. Conclusions We report the largest genome-wide methylation analysis of RPE in AMD along with associated gene expression changes to date, for the first-time reaching genome-wide significance, and identified novel targets for functional and future therapeutic intervention studies. The novel differentially methylated genes SKI and GTF2H4 have not been previously associated with AMD, and regulate disease pathways implicated in AMD, including TGF beta signaling (SKI) and transcription-dependent DNA repair mechanisms (GTF2H4).

Yearly, about 1.5 million people become chronically infected with hepatitis C virus (HCV) and for the 71 million with chronic HCV infection about 400,000 die from related morbidities, including liver cirrhosis and cancer. Effective treatments exist, but challenges including cost-of-treatment and wide-spread undiagnosed infection, necessitates the development of vaccines. Vaccines should induce neutralizing antibodies (NAbs) against the HCV envelope (E) transmembrane glycoprotein 2, E2, which partly depends on its interaction partner, E1, for folding. Here, we generated three soluble HCV envelope protein antigens with the transmembrane regions deleted (i.e., fused peptide backbones), termed sE1E2 (E1 followed by E2), sE2E1 (E2 followed by E1), and sE21E (E2 followed by inverted E1). The E1 inversion for sE21E positions C-terminal residues of E1 near C-terminal residues of E2, which is in analogy to how they likely interact in native E1/E2 complexes. Probing conformational E2 epitope binding using HCV patient-derived human monoclonal antibodies, we show that sE21E was superior to sE2E1, which was consistently superior to sE1E2. This correlated with improved induction of NAbs by sE21E compared with sE2E1 and especially compared with sE1E2 in female BALB/c mouse immunizations. The deletion of the 27 N-terminal amino acids of E2, termed hypervariable region 1 (HVR1), conferred slight increases in antigenicity for sE2E1 and sE21E, but severely impaired induction of antibodies able to neutralize in vitro viruses retaining HVR1. Finally, comparing sE21E with sE2 in mouse immunizations, we show similar induction of heterologous NAbs. In summary, we find that C-terminal E2 fusion of E1 or 1E is superior to N-terminal fusion, both in terms of antigenicity and the induction of heterologous NAbs. This has relevance when designing HCV E1E2 vaccine antigens.

PurposeTo determine whether age-related macular degeneration (AMD) severity or the frequency of retinal pigment epithelium mitochondrial DNA lesions differ in human donor eyes that have undergone cataract surgery compared to phakic eyes.MethodsEyes from human donors aged ≥ 55 years were obtained from the Minnesota Lions Eye Bank. Cataract surgery status was obtained from history provided to Eye Bank personnel by family members at the time of tissue procurement. Donor eyes were graded for AMD severity using the Minnesota Grading System. Quantitative PCR was performed on DNA isolated from macular punches of retinal pigment epithelium to quantitate the frequency of mitochondrial DNA lesions in the donor tissue. Univariable and multivariable analyses were performed to evaluate for associations between (1) cataract surgery and AMD severity and (2) cataract surgery and mitochondrial DNA lesion frequency.ResultsA total of 157 subjects qualified for study inclusion. Multivariable analysis with age, sex, smoking status, and cataract surgery status showed that only age was associated with AMD grade. Multivariable analysis with age, sex, smoking status, and cataract surgery status showed that none of these factors were associated with retinal pigment epithelium mitochondrial DNA lesion frequency.ConclusionsIn this study of human donor eyes, neither retinal pigment epithelium mitochondrial DNA damage nor the stage of AMD severity are independently associated with cataract surgery after adjusting for other AMD risk factors. These new pathologic and molecular findings provide evidence against a relationship between cataract surgery and AMD progression and support the idea that cataract surgery is safe in the setting of AMD.

Yellow Rail (Coturnicops noveboracensis) is a species of conservation concern, while the Eastern Black Rail (Laterallus jamaicensis jamaicensis) has recently been listed as Threatened. Both species winter primarily in coastal areas from Texas through North Carolina. Between-year winter site fidelities of these species have not previously been reported, so we analyzed the recapture histories of Yellow Rails banded in southeastern Oklahoma (from 2008 to 2018) and coastal Texas (from 2006 to 2018), as well as Black Rails banded in Texas from 2006 to 2018. We banded 111 Yellow Rails in Oklahoma and 421 Yellow Rails in Texas during this time, as well as 94 Black Rails in Texas. Although Yellow Rails were routinely recaptured within season, only five individuals were recaptured between years. All recaptured birds were in Texas. Black Rails likewise exhibited low interannual site fidelity, with no interannual recaptures. The apparent low interannual site fidelity of Yellow Rails (1.1% in Texas, 0% in Oklahoma) and Black Rails (0%) is similar to that reported for other grassland species wintering in grasslands in this area (e.g., LeConte’s Sparrow (Ammodramus leconteii), Henslow’s Sparrow (A. henslowii), and Sedge Wren (Cistothorus stellaris)), and we speculate that low site fidelity could potentially be advantageous for species that use early successional habitats.

Anxiety disorders are the most common mental illness in the U.S. and are estimated to consume one-third of the country’s mental health spending. Although anxiolytic therapies are available, many patients exhibit treatment-resistance, relapse, or substantial side effects. An urgent need exists to explore the underlying mechanisms of chronic anxiety and to develop alternative therapies. Presently, we identified dihydromyricetin (DHM), a flavonoid that has anxiolytic properties in a mouse model of isolation-induced anxiety. Socially isolated mice demonstrated increased anxiety levels and reduced exploratory behavior measured by elevated plus-maze and open-field tests. Socially isolated mice showed impaired GABAergic neurotransmission, including reduction in GABAA receptor-mediated extrasynaptic tonic currents, as well as amplitude and frequency of miniature inhibitory postsynaptic currents measured by whole-cell patch-clamp recordings from hippocampal slices. Furthermore, intracellular ATP levels and gephyrin expression decreased in anxious animals. DHM treatment restored ATP and gephyrin expression, GABAergic transmission and synaptic function, as well as decreased anxiety-like behavior. Our findings indicate broader roles for DHM in anxiolysis, GABAergic neurotransmission, and synaptic function. Collectively, our data suggest that reduction in intracellular ATP and gephyrin contribute to the development of anxiety, and represent novel treatment targets. DHM is a potential candidate for pharmacotherapy for anxiety disorders.

Retinal pigment epithelium (RPE) transplantation for the treatment of macular degeneration has been studied for over 30 years. Human clinical trials have demonstrated that RPE monolayers exhibit improved cellular engraftment and survival compared to single cell suspensions. The use of a scaffold facilitates implantation of a flat, wrinkle-free, precisely placed monolayer. Scaffolds currently being investigated in human clinical trials are non-degradable which results in the introduction of a chronic foreign body. To improve RPE transplant technology, a degradable scaffold would be desirable. Using human fibrin, we have generated scaffolds that support the growth of an RPE monolayer in vitro. To determine whether these scaffolds are degraded in vivo, we developed a surgical approach that delivers a fibrin hydrogel implant to the sub-retinal space of the pig eye and determined whether and how fast they degraded. Using standard ophthalmic imaging techniques, the fibrin scaffolds were completely degraded by postoperative week 8 in 5 of 6 animals. Postmortem histologic analysis confirmed the absence of the scaffold from the subretinal space at 8 weeks, and demonstrated the reattachment of the neurosensory retina and a normal RPE-photoreceptor interface. When mechanical debridement of a region of native RPE was performed during implantation surgery degradation was accelerated and scaffolds were undetectable by 4 weeks. These data represent the first in situ demonstration of a fully biodegradable scaffold for use in the implantation of RPE and other cell types for treatment of macular degeneration and other retinal degenerative diseases.

Recent efforts have been made to develop hot-air-dried chips as a value-added product for persimmon fruit (Diospyros kaki). However, the long-term quality of this product and its dependence on processing conditions and packaging type have not yet been explored. Hence, in this work, chips were prepared from “Hachiya” persimmon fruit in 2 ways: 2 mm thickness fruit slices, hot-air-dried for 5 hours (Preparation 2–5), and 6 mm slices, dried for 10 hours (Preparation 6–10). The dried chips were then packaged into 2 bag types (plastic zip-top and metallized polyethylene terephthalate (Met-PET) with desiccant packets), stored under ambient conditions, and sampled for moisture-related, microbial, texture, and chemical quality traits throughout the 1-year storage study. It was found that–with the exception of a marked decrease in ascorbic acid (vitamin C)—all 4 Preparation/Packaging combinations yielded products that maintained acceptable color, microbial, nutritional, and textural quality throughout the entire study. Compared to Preparation 2–5 chips, Preparation 6–10 chips generally had lower contents of healthy nutrients—antioxidant compounds, ascorbic acid, and carotenoids. Evidence of invertase activity was found for Preparation 6–10 chips but not for Preparation 2–5 chips. For both preparations, the contents of ascorbic acid and carotenoids experienced time-dependent decreases. Plastic zip-top bags and Met-PET bags with desiccant packets performed equally well as packaging for most of the quality metrics tracked during this study. Overall, this work has demonstrated that hot-air-drying of persimmon slices is a promising approach to create a value-added product with at least one year of shelf-life.

Abstract GABAergic inhibitory transmission is involved in the acute and chronic effects of ethanol on the brain and behavior. One-dose ethanol exposure induces transient plastic changes in GABAA receptor subunit levels, composition, and regional and subcellular localization. Rapid down-regulation of early responder δ subunit-containing GABAA receptor subtypes mediating ethanol-sensitive tonic inhibitory currents in critical neuronal circuits corresponds to rapid tolerance to ethanol’s behavioral responses. Slightly slower, α1 subunit-containing GABAA receptor subtypes mediating ethanol-insensitive synaptic inhibition are down-regulated, corresponding to tolerance to additional ethanol behaviors plus cross-tolerance to other GABAergic drugs including benzodiazepines, anesthetics, and neurosteroids, especially sedative-hypnotic effects. Compensatory up-regulation of synaptically localized α4 and α2 subunit-containing GABAA receptor subtypes, mediating ethanol-sensitive synaptic inhibitory currents follow, but exhibit altered physio-pharmacology, seizure susceptibility, hyperexcitability, anxiety, and tolerance to GABAergic positive allosteric modulators, corresponding to heightened alcohol withdrawal syndrome. All these changes (behavioral, physiological, and biochemical) induced by ethanol administration are transient and return to normal in a few days. After chronic intermittent ethanol (CIE) treatment the same changes are observed but they become persistent after 30 or more doses, lasting for at least 120 days in the rat, and probably for life. We conclude that the ethanol-induced changes in GABAA receptors represent aberrant plasticity contributing critically to ethanol dependence and increased voluntary consumption. We suggest that the craving, drug-seeking, and increased consumption in the rat model are tied to ethanol-induced plastic changes in GABAA receptors, importantly the development of ethanol-sensitive synaptic GABAA receptor-mediating inhibitory currents that participate in maintained positive reward actions of ethanol on critical neuronal circuits. These probably disinhibit nerve endings of inhibitory GABAergic neurons on dopamine reward circuit cells, and limbic system circuits mediating anxiolysis in hippocampus and amygdala. We further suggest that the GABAA receptors contributing to alcohol dependence in the rat and presumably in human alcohol use disorders (AUD) are the ethanol-induced up-regulated subtypes containing α4 and most importantly α2 subunits. These mediate critical aspects of the positive reinforcement of ethanol in the dependent chronic user while alleviating heightened withdrawal symptoms experienced whenever ethanol is absent. The speculative conclusions based on firm observations are readily testable.

Abstract Background To describe a case of von Hippel-Lindau disease with peripheral retinal nonperfusion. Case presentation A 66-year-old female with known cerebellar and midbrain hemangioblastomas was evaluated for a retinal hemangioblastoma in the right eye. She underwent widefield fluorescein angiography, which showed hyperfluorescence localized to the hemangioblastoma surrounded by peripheral retinal nonperfusion in the same quadrant. Conclusions Further widefield imaging studies are required to determine if peripheral retinal nonperfusion is a common finding in von Hippel-Lindau disease.

The enzyme ADP-glucose pyrophosphorylase (ADP-Glc PPase) controls the biosynthesis of glycogen in bacteria and starch in plants. It is regulated by various activators in different organisms according to their metabolic characteristics. In Escherichia coli, the major allosteric activator is fructose 1,6-bisphosphate (FBP). Other potent activator analogs include 1,6-hexanediol bisphosphate (HBP) and pyridoxal 5′-phosphate (PLP). Recently, a crystal structure with FBP bound was reported (PDB ID: 5L6S). However, it is possible that the FBP site found is not directly responsible for the activation of the enzyme. We hypothesized FBP activates by binding one of its phosphate groups to another site (“P1”) in which a sulfate molecule was observed. In the E. coli enzyme, Arg40, Arg52, and Arg386 are part of this “P1” pocket and tightly complex this sulfate, which is also present in the crystal structures of ADP-Glc PPases from Agrobacterium tumefaciens and Solanum tuberosum. To test this hypothesis, we modeled alternative binding conformations of FBP, HBP, and PLP into “P1.” In addition, we performed a scanning mutagenesis of Arg residues near potential phosphate binding sites (“P1,” “P2,” “P3”). We found that Arg40 and Arg52 are essential for FBP and PLP binding and activation. In addition, mutation of Arg386 to Ala decreased the apparent affinity for the activators more than 35-fold. We propose that the activator binds at this “P1” pocket, as well as “P2.” Arg40 and Arg52 are highly conserved residues and they may be a common feature to complex the phosphate moiety of different sugar phosphate activators in the ADP-Glc PPase family.