Search

David PW Rastall,1 Andrea Amalfitano1,2 1Department of Microbiology and Molecular Genetics, 2Department of Pediatrics, College of Osteopathic Medicine, Michigan State University, East Lansing, MI, USA Abstract: Lysosomal storage disorders (LSDs) are a group of genetic diseases that result in metabolic derangements of the lysosome. Most LSDs are due to the genetic absence of a single catabolic enzyme, causing accumulation of the enzyme's substrate within the lysosome. Over time, tissue-specific substrate accumulations result in a spectrum of symptoms and disabilities that vary by LSD. LSDs are promising targets for gene therapy because delivery of a single gene into a small percentage of the appropriate target cells may be sufficient to impact the clinical course of the disease. Recently, there have been several significant advancements in the potential for gene therapy of these disorders, including the first human trials. Future clinical trials will build upon these initial attempts, with an improved understanding of immune system responses to gene therapy, the obstacle that the blood–brain barrier poses for neuropathic LSDs, as well other biological barriers that, when overcome, may facilitate gene therapy for LSDs. In this manuscript, we will highlight the recent innovations in gene therapy for LSDs and discuss the clinical limitations that remain to be overcome, with the goal of fostering an understanding and further development of this important field. Keywords: human trials, clinical trials, gene therapy, lysosomal storage disease, blood-brain barrier, adeno-associated virus, lentivirus, adenovirus

ABSTRACT: Cheese presents extensive variability in physical, chemical, and sensory characteristics according to the variety of processing methods and conditions used to create it. Relationships between the many characteristics of cheeses are known for single cheese types or by comparing a few of them, but not for a large number of cheese types. This case study used the properties recorded on 1,050 different cheeses from 107 producers grouped into 37 categories to analyze and quantify the interrelationships among the chemical and physical properties of many cheese types. The 15 cheese traits considered were ripening length, weight, firmness, adhesiveness, 6 different chemical characteristics, and 5 different color traits. As the 105 correlations between the 15 cheese traits were highly variable, a multivariate analysis was carried out. Four latent explanatory factors were extracted, representing 86% of the covariance matrix: the first factor (38% of covariance) was named Solids because it is mainly linked positively to fat, protein, water-soluble nitrogen, ash, firmness, adhesiveness, and ripening length, and negatively to moisture and lightness; the second factor (24%) was named Hue because it is linked positively to redness/blueness, yellowness/greenness, and chroma, and negatively to hue; the third factor (17%) was named Size because it is linked positively to weight, ripening length, firmness, and protein; and the fourth factor (7%) was named Basicity because it is linked positively to pH. The 37 cheese categories were grouped into 8 clusters and described using the latent factors: the Grana Padano cluster (characterized mainly by high Size scores); hard mountain cheeses (mainly high Solids scores); very soft cheeses (low Solids scores); blue cheeses (high Basicity scores), yellowish cheeses (high Hue scores), and 3 other clusters (soft cheeses, pasta filata and treated rind, and firm mountain cheeses) according to specific combinations of intermediate latent factors and cheese traits. In this case study, the high variability and interdependence of 15 major cheese traits can be substantially explained by only 4 latent factors, allowing us to identify and characterize 8 cheese type clusters.

ABSTRACT: Rotational crossbreeding has not been widely studied in relation to the enteric methane emissions of dairy cows, nor has the variation in emissions during lactation been modeled. Milk infrared spectra could be used to predict proxies of methane emissions in dairy cows. Therefore, the objective of this work was to study the effects of crossbreeding on the predicted infrared proxies of methane emissions and the variation in the latter during lactation. Milk samples were taken once from 1,059 cows reared in 2 herds, and infrared spectra of the milk were used to predict milk fat (mean ± SD; 3.79 ± 0.81%) and protein (3.68 ± 0.36%) concentrations, yield (21.4 ± 1.5 g/kg dry matter intake), methane intensity (14.2 ± 2.0 g/kg corrected milk), and daily methane production (358 ± 108 g/d). Of these cows, 620 were obtained from a 3-breed (Holstein, Montbéliarde, and Viking Red) rotational mating system, and the rest were purebred Holsteins. Milk production data and methane traits were analyzed using a nonlinear model that included the fixed effects of herd, genetic group, and parity, and the 4 parameters (a, b, c, and k) of a lactation curve modeled using the Wilmink function. Milk infrared spectra were found to be useful for direct prediction of qualitative proxies, such as methane yield and intensity, but not quantitative traits, such as daily methane production, which appears to be better estimated (450 ± 125 g/d) by multiplying a measured daily milk yield by infrared-predicted methane intensity. Lactation modeling of methane traits showed daily methane production to have a zenith curve, similar to that of milk yield but with a delayed peak (53 vs. 37 d in milk), whereas methane intensity is characterized by an upward curve that increases rapidly during the first third of lactation and then slowly till the end of lactation (10.5 g/kg at 1 d in milk to 15.2 g/kg at 300 d in milk). However, lactation modeling was not useful in explaining methane yield, which is almost constant during lactation. Lastly, the methane yield and intensity of cows from 3-breed rotational crossbreeding are not greater, and their methane production is lower than that of purebred Holsteins (452 vs. 477 g/d). Given the greater longevity of crossbred cows, and their lower replacement rate, rotational crossbreeding could be a way of mitigating the environmental impact of milk production.

Local sheep breeds in the Italian eastern Alps passed from ex situ to in situ conservation. These breeds are mainly reared by smallholders in low-input farming systems. To allow the sustainable use of genetic resources, the economic sustainability of farmers must be supported through production guidelines. Analyzing meat and milk composition and fatty acid profile, we aimed to characterize their products based on breed and diet to identify tailormade sales strategies. Results showed that both meat and milk have good nutritional values and can benefit from a pasture-based diet, irrespective of the breed. These results support the redaction of production guidelines based on the peculiar characteristics of these breeds: being multi-purpose breeds adapted to mountain areas and to grazing, thus contributing to the conservation of cultural and landscape heritage.

The plastisphere, a unique microbial biofilm community colonizing plastic debris and microplastics (MPs) in aquatic environments, has attracted increasing attention owing to its ecological and public health implications. This review consolidates current state of knowledge on freshwater plastisphere, focussing on its biodiversity, community assembly, and interactions with environmental factors. Current biomolecular approaches revealed a variety of prokaryotic and eukaryotic taxa associated with plastic surfaces. Despite their ecological importance, the presence of potentially pathogenic bacteria and mobile genetic elements (i.e., antibiotic resistance genes) raises concerns for ecosystem and human health. However, the extent of these risks and their implications remain unclear. Advanced sequencing technologies are promising for elucidating the functions of plastisphere, particularly in plastic biodegradation processes. Overall, this review emphasizes the need for comprehensive studies to understand plastisphere dynamics in freshwater and to support effective management strategies to mitigate the impact of plastic pollution on freshwater resources.

Modern cetaceans are considered the best anatomical and ecological analogue for many Mesozoic secondary aquatic reptiles. Such similarities extend also after the death of these phylogenetically distant amniotes, when the sinking and decomposition of large carcasses in marine environments (deadfalls) follow common biostratinomic processes. Most taphonomic studies on Mesozoic deadfalls have been limited to shallow-water settings, often neglecting deeper waters. Here we provide a detailed taphonomic survey of ichthyosaurs, pliosaurs and metriorhynchoids from the pelagic Middle-Upper Jurassic Rosso Ammonitico Veronese (RAV) of northeastern Italy. Our taphonomic revision of the RAV tetrapod record highlights a common poor state of preservation of the bones, often associated with abundant macrofossils, consistent with a prolonged exposure of carcasses on a well-oxygenated seafloor. For the first time we confirm the role of nautiloids as active mobile scavengers by means of tens of beak elements found closely associated with, or even piercing, the bones. Hexanchiform shark teeth are also found associated with the carcasses, supporting a distinctive deep-water mobile scavenging community. Echinoids, sponges and other bioeroders are identified as representative of the enrichment-opportunist stage, and a high concentration of belemnites is believed to be indicative of mass-spawning deaths in the surroundings of the carcasses. Abundant crinoids are recognized as part of the reef stage by colonization of the eroded bones. While some of our data deviate from previous Mesozoic reptile-falls from shallow-waters, they are consistent with findings at Recent whale-falls in bathyal zones, and overall represent a precious window into the complex ecology of Jurassic open seas.

Abstract Background Induction of potent long lasting effector T cell responses against liver stage malaria antigens strongly correlates with protection from malaria. While Adenovirus serotype 5 (Ad5) based malaria vaccine platforms have the ability to induce potent effector T cell responses against transgenes, high rates of pre-existing Ad5 immunity in malaria endemic regions has prompted study of alternative Ad serotype based malaria vaccines as replacements for Ad5 based malaria vaccines. The research described in this article examines the utility of alternative serotype adenovirus serotype 4 (Ad4) expressing a sporozoite surface protein (circumsporozoite protein (CSP)) (Ad4-CSP) to induce immune responses against CSP. The immunogenicity of Ad4-CSP was also tested in homologous and heterologous prime boost vaccinations in both Ad5 naïve and Ad5 immune backgrounds as compared to use of Ad5-CSP. Results In Ad5 naïve animals, use of Ad4-CSP priming vaccinations followed by boosting with Ad5-CSP (Ad4-CSP/Ad5-CSP) maximally increased the numbers of CSP specific cytokine secreting cytotoxic T cells relative to repeated use of Ad5-CSP. The Ad4-CSP/Ad5-CSP regimen also induced equivalent levels of CSP specific cell killing as did homologous prime-boost vaccinations with Ad5-CSP, despite stimulating lower numbers of CSP specific cytotoxic T cells. Priming with Ad4-CSP followed by a homologous boost resulted in significantly less CSP specific humoral responses than any other vaccination regimen tested in Ad naïve animals. In Ad5 immune animals, addition of Ad4-CSP in homologous or heterologous prime boost resulted in inductions of higher CSP specific responses than animals repeatedly vaccinated with Ad5-CSP alone. However, the observed responses were well below those observed in similarly treated Ad naïve mice. Conclusions While the Ad4-CSP/Ad5-CSP and Ad5-CSP/Ad5-CSP vaccination regimens resulted in equivalent CSP specific killing in Ad naïve animals, Ad4-CSP/Ad5-CSP achieved this result with a lower percentage of CSP specific CD8+ T cells and a higher number of IFNγ secreting cells, suggesting that the Ad4-CSP/Ad5-CSP vaccination regimen elicits more efficient cytotoxic T cells. In Ad5 immune animals use of Ad4-CSP improved CSP specific immune responses as compared to repeated use of Ad5-CSP, but could not achieve the levels of immunogenicity observed when the same vaccine regimens were used in Ad naïve animals. These data indicate the existence of some level of immunological cross-reactivity between these two adenovirus subgroups. Based on these results, it is suggested that future studies should undertake similarly stringent analyses of alternative Ad serotypes to establish their effectiveness as replacements for Ad5.

Semi-articulated remains of a large chelonioid turtle from the Turonian strata (Upper Cretaceous; ca. 93.9-89.8 Myr) near Sant'Anna d'Alfaedo (Verona province, northeastern Italy) are described for the first time. Together with the skeletal elements, the specimen also preserves pebbles inside the thoracic area which are lithologically distinct from the surrounding matrix. These allochthonous clasts are here interpreted as geo-gastroliths, in-life ingested stones that resided in the digestive tract of the animal. This interpretation marks the first reported evidence of geophagy in a fossil marine turtle. SEM-EDS analysis, together with macroscopic petrological characterization, confirm the presence of both siliceous and carbonatic pebbles. These putative geo-gastroliths have morphometries and size ranges more similar to those of gastroliths in different taxa (fossils and extant) than allochthonous "dropstone" clasts from the same deposit that were carried by floating vegetation A dense pitted pattern of superficial erosion is microscopically recognizable on the carbonatic gastroliths, consistent with surface etching due to gastric acids. The occurrence of a similar pattern was demonstrated by the experimental etching of carbonatic pebbles with synthetic gastric juice. Gut contents of modern green sea turtles (Chelonia mydas) were surveyed for substrate ingestion, providing direct evidence of geophagic behavior in extant chelonioids. Comparison with modern turtle dietary habits may suggests that the pebbles were ingested as a way to supplement calcium after or in preparation for egg deposition, implying that the studied specimen was possibly a gravid female.

IntroductionOnce dispersed in water, plastic materials become promptly colonized by biofilm-forming microorganisms, commonly known as plastisphere.MethodsBy combining DNA sequencing and Confocal Laser Scanning Microscopy (CLSM), we investigated the plastisphere colonization patterns following exposure to natural lake waters (up to 77 days) of either petrochemical or biodegradable plastic materials (low density polyethylene - LDPE, polyethylene terephthalate - PET, polylactic acid - PLA, and the starch-based MaterBi® - Mb) in comparison to planktonic community composition. Chemical composition, water wettability, and morphology of plastic surfaces were evaluated, through Transform Infrared Spectroscopy (ATR-FTIR), Scanning Electron Microscopy (SEM), and static contact angle analysis, to assess the possible effects of microbial colonization and biodegradation activity.Results and DiscussionThe phylogenetic composition of plastisphere and planktonic communities was notably different. Pioneering microbial colonisers, likely selected from lake waters, were found associated with all plastic materials, along with a core of more than 30 abundant bacterial families associated with all polymers. The different plastic materials, either derived from petrochemical hydrocarbons (i.e., LDPE and PET) or biodegradable (PLA and Mb), were used by opportunistic aquatic microorganisms as adhesion surfaces rather than carbon sources. The Mb-associated microorganisms (i.e. mostly members of the family Burkholderiaceae) were likely able to degrade the starch residues on the polymer surfaces, although the Mb matrix maintained its original chemical structure and morphology. Overall, our findings provide insights into the complex interactions between aquatic microorganisms and plastic materials found in lake waters, highlighting the importance of understanding the plastisphere dynamics to better manage the fate of plastic debris in the environment.

The Somma–Vesuvius volcanic complex emitted huge quantities of volcanic materials over a period from before 18,300 years BP to 1944. The activity during the last period, from post-AD 1631 to 1944, primarily produced lava and pyroclastics via effusive and strombolian eruptions. We investigated the pedogenesis on rocks formed from post-AD 1631 to 1944, occurring on the slopes of Mt. Vesuvius up to Gran Cono Vesuviano and in the northern valley separating Vesuvius from the older Mt. Somma edifice. Pertinent morphological, physical, chemical, and mineralogical (XRD and FT-IR) soil properties were studied. The results indicated the existence of thin and deep stratified soils on lava, as well as the presence of loose detritic covers formed via pyroclastic emplacement and redistribution. The soils showed minimal profile differentiation, frequently with layering recording the episodic addition of sediments. We found that the dominant coarse size of primary mineral particles was preserved, and there was a low level of clay production. The main mineralogical assemblage present in sands also persisted in clays, indicating the physical breaking of the parent material. Chemical weathering produced mineral modifications towards the active forms of Al and Fe and was also attested in selected soils by glass alteration, allophane production, and the presence of analcime in clay as a secondary product from leucite. The differences in glass alteration and analcime production found in the selected soils on lava were related to soil particle size and soil thickness. Concerning the youngest soil present on Gran Cono Vesuviano, other factors, such as the substratum’s age and site elevation, appeared to be implicated.

The transformation of milk into cheese largely depends on the technological properties of the raw material, with breed being a crucial factor that influences both the composition and coagulation properties of the milk used for cheesemaking. This study uses canonical correlation analysis to explore the relationships between physicochemical traits and coagulation properties in milk from various Spanish breeds, aiming to identify both common and breed-specific patterns that impact milk technological aptitude. A total of 832 milk samples from Manchega, Assaf, Merino de Grazalema, and Merino de Los Pedroches breeds were analyzed. The milk characteristics investigated included pH, composition (fat, protein, lactose, total solids), and coagulation properties (curd firmness—A60, rennet clotting time—RCT, curd firming time—k20, and individual laboratory curd yield—ILCY). The results reveal a shared correlation structure across breeds and unique covariation patterns in some breeds that deviate from the general trend. While Assaf and Merino de Los Pedroches follow the common correlation pattern, Manchega and Merino de Grazalema exhibit distinct patterns. This research underscores the need for in-depth study and suggests that the dairy industry could benefit from shifting from the traditional focus on maximizing fat and protein for higher curd yields to considering technological traits for selective breeding.

Sheep milk from local breeds is important for the production of high-quality cheeses throughout the Mediterranean region, such as Manchego cheese in Spain. To maintain sustainable and efficient production, it is necessary to reach a better understanding of how the composition and hygiene of the milk affect the coagulation process, with the aim of optimizing production yield. This study implemented a stochastic production frontier function to estimate the potential production of curd and efficiency using data from the four seasons of a study of 77 Manchega sheep farms. The Cobb–Douglas production frontier model was estimated using the maximum likelihood estimation method. The results showed that the content of protein, lactose, and fat exhibited increasing returns to scale, with protein content being the most significant factor for curd production. Approximately half of the inefficiency was due to factors related to the technological properties and the hygiene of the milk. The pH, curd firmness, and concentration of lactic acid bacteria improved the efficiency of coagulation, while the concentration of spores of lactate-fermenting Clostridium spp., Pseudomonas spp., staphylococci, and catalase-negative gram-positive cocci favored the inefficiency of the coagulation process. To date, this is the first study to evaluate the effect of different factors, such as microbial groups, milk composition, and technological properties, on the efficiency of the coagulation process in dairy sheep.

This study conducted a seasonal analysis of bulk tank milk from 77 sheep farms to establish relationships between the concentration of major microbial groups and milk coagulation properties. The investigated milk traits included composition (pH, fat, casein, lactose), coagulation properties (curd firmness: A60-, rennet clotting time: RCT-, curd firming time: k20-, curd yield: CY-), and somatic cell score (SCS). The main microbial groups analyzed were total mesophilic bacteria (SPC), thermodurics (THERMO), psychrotrophs (PSYCHRO), Pseudomonas spp. (PSEUDO), lactic acid bacteria (LAB), catalase-negative gram-positive cocci (GPCNC), Escherichia coli (ECOLI), coliforms other than Escherichia coli (COLI), coagulase-positive staphylococci (CPS), coagulase-negative staphylococci (CNS), and spores of lactate-fermenting Clostridium (BAB). Mixed linear models were used to explore associations between coagulation properties and the aforementioned variables. Results demonstrated that incorporating microbial loads into the models improves their fit and the relative quality of the outcomes. An important seasonality is demonstrated by an increase in CY and A60, along with a decrease in RCT and k20 during autumn and winter, contrasting with spring and summer. BAB concentration resulted in a reduction of A60 and an increase in RCT, whereas SPC concentration led to an enhancement of A60 and a reduction in RCT. An increase in GPCNC concentration was associated with an increase in k20 and a decrease in CY.

A new lower tooth plate of Ptychodus decurrens from the Turonian (Upper Cretaceous) of the Dalmatian region (southern Croatia) is documented here for the first time. The specimen represents the first articulated dentition of a ptychodontid shark that has been discovered from the Balkan Peninsula and the most complete ever found for the un-cuspidate species P. decurrens up to now. The reconstruction of the entire lower dentition of P. decurrens based on this exceptionally well-preserved dentition shows a wider crushing plate than previously hypothesised with bulgy teeth limited to the central area. Even though a defined cusp is missing, the occlusal surface of the teeth is undeniably raised and bulgy in some un-cuspidate species of Ptychodus (e.g., P. decurrens). This compels us to reconsider the use of terms such as high- and low-crowned as well as cuspidate and un-cuspidate. In addition, specimens previously assigned to dubious species (P. depressus, P. levis and P. oweni) or even varieties (P. polygyrus var. sulcatus and P. decurrens var. multiplicatus) are reassigned here to P. decurrens based on a careful comparison of the type materials. The reassessment of tooth root morphologies provides indicative traits for the identification of different genera of ptychodontid sharks (Paraptychodus and Ptychodus). The taxonomic revision presented here is crucial for securing a stable taxonomy and systematics of the shell-crushing shark P. decurrens, as well as of all ptychodontid sharks. The resulting updated taxonomy, together with the description and reconstruction of the new crushing plate, greatly contribute to a better understanding of one of the most enigmatic families (Ptychodontidae) of Mesozoic elasmobranchs. The detailed investigation of the new dentition of Ptychodus from Dalmatia is also a further step towards the discovery of Upper Cretaceous ichthyofaunas of one of the most palaeontologically important areas of the Balkan Peninsula. [ABSTRACT FROM AUTHOR]

The aim of this study was to characterize the highly detailed fatty acid (FA) profiles of 258 cheeses of 18 different categories of cheese collected in the mountains and on the plains of the Veneto region (Italy). The results clearly showed that, aside from the distinctive FA profiles of goat cheeses (more short-chain FAs and fewer MUFAs), the three categories of Formaggio di Malga (artisanal cheeses produced on temporary summer farms on Alpine pastures where transhumance is practiced) were very different from the other cheese categories in terms of their much higher CLA and omega-3 contents. Two categories of cheese from permanent farms in the mountains (Morlacco del Grappa and Monte Veronese PDO) were intermediate, and two other categories of cheeses originating in the mountains (Asiago PDO and Montasio PDO), but now produced mainly on the plains, were not distinguishable from the other cheese categories. The very detailed profile (65 individual FA, 11 isomers, and 12 groups of FAs) and the large number of cheese types analyzed (18) may represent a useful reference for future investigations, especially on the causes of variability in FAs and on their relationships with sensory properties and nutrition/health in humans. [ABSTRACT FROM AUTHOR]

Milk from cows and buffaloes represents the major source of dairy products worldwide. Cheese is consumed fresh, ripened, cooked, or as recipe ingredient in many different types. Using Paneer and Tosella/Schiz model cheeses as a case study of cooked cheeses, in this short communication we evaluated the cheese yield and quality of fresh cheese before and after cooking comparing the two major dairy species (cows and buffaloes). A total of 75 model cheeses (37 Paneer and 38 Tosella/Schiz) were made. Slices of all the cheeses were cooked on a griddle at 130 °C. Milk, whey, and cheese composition traits, cheese-making efficiency traits, and cooked cheese quality traits were evaluated using a mixed model that included the fixed effects of cheese type and animal species, and their interaction, and the random effects of session, animal within species, and the residual. Higher cheese yields are obtained from the buffalo milk not only because more concentrated in nutrients, but also for the greater recovery of nutrients in cheese. There was a greater recovery of protein with the acid/heat Paneer procedure than the rennet Tosella/Schiz procedure, but we found a species × cheese type interaction for fresh cheese yield. The interaction was related mainly to the higher water retained in the Tosella/Schiz curd rather than to total solids. This led also to greater cooking weight-loss compared with Paneer. The qualitative traits of cheese were highly affected by griddling. Future research on the sensorial perception of cooked cheeses from the two major dairy species is needed to predict consumer response. HIGHLIGHTS: Cow and buffalo milk yield different results using different cheese-making procedures for producing cooked cheese. Heat/acid coagulation, like for Paneer, allows for the recovery of the majority of the fat, almost all the caseins in the curd, and about half the whey proteins. Mild temperature/rennet coagulation procedure, like for Tosella/Schiz, increases water retention in fresh cheese and weight loss during cooking. The cheese-making procedures developed for the milk of one species cannot be transferred directly to the milk of another species because of an interaction between species and the cheese-making procedure. [ABSTRACT FROM AUTHOR]