BACKGROUND: A freeze-dried, platelet-derived hemostatic agent (FPH) was developed for acute hemorrhage. The canine product (cFPH) was developed for use in preclinical models supporting human product (hFPH) investigations.
METHODS: A carotid artery bypass graft (CABG) study in dogs compared 3 dosages of cFPH to canine liquid stored platelets (cLSP) and vehicle (VEH) control groups. Histopathological analysis and blood loss assessments were completed. A separate ex-vivo synthetic graft study assessed thrombogenicity via blood from human and canine donors that was combined with species-specific FPH or apheresis platelets. Characterization of cFPH and hFPH included thrombin generation, total thrombus formation, and scanning electron microscopy.
RESULTS: Blood loss was reduced in CABG dogs receiving standard of care (cLSP) or cFPH treatment compared to VEH control; a cFPH dose effect signal was observed. Further, cFPH dosing up to 5 × 109 cells/kg was not associated with increased mortality or occlusion of the anastomosis sites, and histopathologic evidence of off-target thrombosis was not detected. When passed through a synthetic graft (ex vivo), whole blood combined with species-specific FPH did not result in thrombosis beyond that of whole blood control. In vitro testing and imaging of cFPH and FPH were comparable.
CONCLUSIONS: A single dose of cFPH or cLSP reduced blood loss in a pilot surgical study and was well tolerated with no related adverse events. Further, the hemostatic activity and characteristics of cFPH are comparable to that of hFPH, suggesting that research findings from the canine product are likely to inform the development of the human product.

We evaluated the in vitro effects of lyophilization for 2 vesicular stomatitis virus-based vaccines by using 3 stabilizing formulations and demonstrated protective immunity of lyophilized/reconstituted vaccine in guinea pigs. Lyophilization increased stability of the vaccines, but specific vesicular stomatitis virus-based vaccines will each require extensive analysis to optimize stabilizing formulations.

In microbiome studies, fecal and oral samples are stored and processed in different ways, which could affect the observed microbiome composition. In this study, we compared storage and processing methods applied to samples prior to DNA extraction to determine how each affected microbial community diversity as assessed by 16S rRNA gene sequencing. We collected dental swabs, saliva, and fecal samples from 10 individuals, with three technical replicates per condition. We assessed four methods of storing and processing fecal samples prior to DNA extraction. We also compared different fractions of thawed saliva and dental samples to fresh samples. We found that lyophilized fecal samples, fresh whole saliva samples, and the supernatant fraction of thawed dental samples had the highest levels of alpha diversity. The supernatant fraction of thawed saliva samples had the second highest evenness compared to fresh saliva samples. Then, we investigated the differences in observed community composition at the domain and phylum levels and identified the amplicon sequence variants (ASVs) that significantly differed in relative abundance between the conditions. Lyophilized fecal samples had a greater prevalence of Archaea as well as a greater ratio of Firmicutes to Bacteroidetes compared to the other conditions. Our results provide practical considerations not only for the selection of storage and processing methods but also for comparing results across studies. Differences in processing and storage methods could be a confounding factor influencing the presence, absence, or differential abundance of microbes reported in conflicting studies.

This study evaluates the nutritional value of a baked pork meat product containing lyophilized dragon fruit pulp. The selected nutritional properties of a baked pork meat product fortified with lyophilized Hylocereus undatus pulp in doses of 0.5%, 1.5%, 2.5%, and 4% were evaluated. For this assessment, changes in the basic chemical composition of the products, the content of calcium, magnesium, potassium, iron, and phosphorus, and the profile of fatty acids were considered. Additionally, characteristics typical for meat products, such as pH, water activity, oxidation-reduction potential or thiobarbituric acid reactive substances, and antioxidant properties of the product during 21 days of refrigerated storage, were assessed. The findings indicate that the use of higher doses of lyophilizate, i.e., in the amounts of 2.5% and 4%, significantly (p < 0.05) increases the nutritional value of meat products, leading to an increase in the concentration of essential minerals important for the proper functioning of the human body (calcium, magnesium, potassium, and iron). These changes occurred without affecting the basic chemical composition (except for an increase in the content of fat and carbohydrates in the sample with the addition of 4% lyophilizate). The introduction of the fortification treatment improved the fatty acid profile, resulting in an increase in the content of C14:0, C16:0, C20:0, and C20:5n3. In addition, in the variant with a 4% dosage, there was an increased content of C8:0, C10:0, C16:1n7, C18:0, C18:1n9C + C18:1n9t, and C18:2n6C + C18:2n6t, C18:3n3 (alpha), C20:1n15, and C20:1n9. In this particular variant, an increase in saturated-, monounsaturated-, and polyunsaturated fatty acids was also observed, which was associated with an increased level of TBARS in meat products. However, the increase in the dose of lyophilizate caused an increase in the antiradical effect of meat extracts. Based on the results obtained, it seems reasonable to use a plant additive in the form of lyophilized dragon fruit pulp in the amount of 4.0% in the production of pork meat products.

As the only Food and Drug Administration (FDA)-approved dual-encapsulation liposome injection for treating Acute myeloid leukemia (AML), CPX-351 outperforms the standard chemotherapy treatment \"DA 7 + 3″ in terms of clinical effectiveness. Although research on dual-loaded liposomes has increased in recent years, little attention has been paid to their preparation, which can affect their quality, efficacy, and safety. This study explored various preparation processes to create the cytarabine/daunorubicin co-loaded liposome (the Cyt/Daun liposome) and eventually settled on two methods: the sequential loading approach, thin film hydration-extrusion-copper ion gradient, and the simultaneous encapsulation technique, copper ion gradient-concentration gradient. Different preparation methods resulted in different particle sizes and encapsulation efficiencies; the two aforementioned preparation processes generated dual-loaded liposomes with comparable physicochemical properties. The sequential encapsulation technique was selected for the subsequent research owing to its higher encapsulation efficiency prior to purification; the prepared Cyt/Daun liposomes had small and uniform particle size (108.6 ± 1.02 nm, Polydispersity index (PDI) 0.139 ± 0.01), negative charge (-(60.2 ± 1.15) mV), high drug encapsulation efficiency (Cyt 88.2 ± 0.24 %, Duan 94.2 ± 0.45 %) and good plasma stability. To improve its storage stability, the Cyt/Daun liposome was lyophilized (-40 °C for 4 h, maintained for 130 min, and dried for 1200 min) using sucrose-raffinose (mass ratio 7:3; glycolipid ratio 4:1, w/w) as a lyoprotectant. The lyophilized liposomes were purple cakes, redissolved rapidly with insignificant alterations in particle size and encapsulation efficiency, and possessed well storage stability. The pharmacokinetic and tissue distribution studies demonstrated that the Cyt/Daun liposome could achieve long circulation and maintain synergic proportions of drugs within 24 h, increasing the accumulation of drugs at tumor sites. Furthermore, the in vitro/in vivo pharmacodynamic studies confirmed its good anti-tumor activity and safety.

An electrochemical study of lyophilized blueberry (Vaccinium corymbosum L.) and raspberry (Rubus idaeus L.) fruits was carried out using microparticulate deposits from ethanolic extracts of fruits in contact with different aqueous electrolytes. This voltammetry is dominated by the oxidation of anthocyanins. It was hypothesized that under conditions of electrochemical generation of reactive oxygen species (ROS), new anthocyanin-derived species with high antioxidant capacity are formed over a wide pH range. In the case of ROS-activated anthocyanins, electrochemical oxidation is associated with loss of sugar moieties. Experimental voltammetric and scanning electrochemical microscopy (SECM) imaging data consistent with this hypothesis are presented.

The antihyperglycemic effect of Plicosepalus acaciae (P. acaciae) extract was proven, but it still needs to be formulated into a suitable dosage form. We aimed at preparing an oral stabilized SLNs for P. acaciae with high payload, to be used as powder for reconstitution, filled into capsule or compressed into tablet. SLNs were prepared by emulsion solvent evaporation technique. Preliminary characterization was performed followed by full assessment of the optimized SLNs suspension and/or its lyophilized form: particle size, zeta potential, surface morphology, percentage entrapment efficiency (% EE), DSC, FTIR and in vitro release studies. The optimized SLNs lyophilized formula (F3L) exhibited acceptable compressibility and flowability. The reconstituted F3L showed % sedimentation volume of 91.83 %, re-dispersibility of 95%, viscosity of 764.33 cp, uniform particle size of 30.28 nm as shown by TEM, polydispersity index (PDI) of 0.16, zeta potential of -36.4 mV, % EE of 89.64 % and drug content of 97.69 %. The physical mixture and F3L FTIR spectrum indicated compatibility of components. In vitro release study showed a burst release in lyophilized formulations followed by slow-release, calculated as total phenolic content. Our previously reported work revealed that the total extracts of P. acaciae and SLNs formulations with the greatest lipid content F3s, demonstrated a considerable blood glucose-lowering effect in diabetic rats. The obtained lyophilized SLNs is promising for preparation of a suitable stable dosage form for P. acaciae extract to be used in treatment of diabetes.

A rapid and sensitive COVID-19 diagnostic test is required to aid in the prevention and control of the current COVID-19 pandemic spread. We developed a colorimetric, rapid, and sensitive RT-LAMP assay for the diagnosis of COVID-19 viral infection.
Complete genome sequences of 41 SARS-CoV-2 isolates from Oman were used in this study. Three primer sets (CoV_S1, CoV_S2, CoV_M1) were developed from all Omani SARS-CoV-2 genome sequences available at the time, targeting the spike protein gene and the M gene. The primer set (CoV_S1) was found to be the most sensitive and specific among the three designed sets. The sensitivity and specificity of the assay were compared to that of qRT-PCR. Direct testing of SARS-CoV-2 spiked saliva with the developed assay was evaluated. Lyophilized colorimetric assays were stored at room temperature and 4 °C and their ability to detect positive samples were tested for a period of 8 weeks.
The RT-LAMP assay was validated by testing 145 COVID-19 clinical samples with a sensitivity of 96.9% and specificity of 94.7% when compared to the validated qRT-PCR assay. The assay specificity was tested against SARS-CoV Frankfurt 1 RNA virus and avian coronaviruses as they tested negative with the developed assay. The assay was lyophilized and managed to detect the positive samples colorimetrically when stored at 4 °C for up to 8 weeks.
The assay can be utilized in its current form as a screening assay with the advantages of being simpler, quicker, and cheaper than the qRT-PCR.

Utilities often test the effectiveness of different granular activated carbons (GACs) to determine which is most advantageous for their system. For surface water systems, in particular, the seasonal and annual variability of natural organic matter (NOM) in the source water makes it difficult to benchmark the effectiveness of GACs over multiple contract periods. This study produced stable, lyophilized NOM from the filtered water (FW), i.e., the influent to GAC contactors, which was then reconstituted (Recon) and tested against the FW itself in parallel rapid small-scale column tests (RSSCTs). The results demonstrated nearly identical NOM breakthrough profiles. RSSCTs conducted with both FW and Recon were shown to simulate the full-scale contactor performance well, while similar RSSCTs with regenerated GAC yielded a slightly earlier breakthrough, possibly due to the changes in GAC characteristics during regeneration and grinding. RSSCTs evaluating the removal of microcystin-LR (MC-LR) in the presence of background NOM contained in FW and Recon showed slightly different results, possibly due to the difference in chloride concentrations of these two waters. This work validates that reconstituted lyophilized NOM can be used as a source water surrogate for GAC evaluations when the constituent of interest is NOM, and potentially for other constituents depending upon the influence of additional inorganic constituents that were not evaluated as part of this study.

The usage of canine amniotic membrane (cAM) is mainly of interest in veterinary ophthalmology. Topical formulations of cAM could deliver the beneficial properties of cAM without the need for surgical intervention. The present study aimed to investigate biological compositions of cAM and its extracts, including their corneal wound healing efficacy. In this study, canine amniotic membrane extract (cAME) and lyophilized canine amniotic membrane extract (cAMX) were developed. Bioactive molecules related to corneal wound healing, including hepatocyte growth factor, tissue inhibitor of metalloproteinase-1 and -2, Thrombospondin-1 and Interleukin-1 receptor antagonist were studied at both gene and protein expression levels. Cell viability and wound healing assays were investigated for the possibility of cAME and cAMX as topical applications. The results demonstrated that all of the relevant genes and proteins were detected in cAM, cAME and cAMX. Both cAME and cAMX showed wound healing properties in vitro and cAME at 1.0 mg/mL concentration appeared to have the best healing efficacy. In conclusion, cAME and cAMX generated for topical use provided promising results in the healing of corneal defects.