Functional and morphological changes in diabetic macular edema over the course of anti‐vascular endothelial growth factor treatment

Purpose:  To evaluate macular morphology and function in diabetic macular edema (DME) over the course of intravitreal anti‐vascular endothelial growth factor (VEGF) treatment with Ranibizumab. Methods:  A consecutive series of 39 study eyes with centre‐involving DME were included in this study. In all subjects, best‐corrected visual acuity (BCVA) according ETDRS protocol, fluorescein angiography (FA), microperimetric macular sensitivity (MP) and Spectral Domain optical coherence tomography (SD‐OCT) cross‐sectional scans were obtained before treatment and after 3 monthly applied intravitreal Ranibizumab injections. Six different morphological qualities [IS/OS layer integrity, outer nuclear layer (ONL) cysts, ONL cyst size, inner nuclear layer (INL) cysts, blocking phenomenon and subretinal fluid] were graded of each cross‐sectional OCT scan before and over the course of treatment by two experienced graders. Correlation analyses between functional and morphological parameters were obtained. Results:  Mean BCVA increased from 26 ± 14 to 33 ± 13 letters after 3 consecutive monthly applied Ranibizumab injections (p < 0.001). Central retinal thickness (CRT) decreased from 504 ± 144 to 387 ± 122 μm (p < 0.001). Over the course of treatment, IS/OS continuity improved (index: 0.56 ± 0.52 to 0.43 ± 0.49, Z = ?1.415, p = 0.157), ONL cyst prevalence and size decreased significantly (index: 0.61 ± 0.44 to 0.56 ± 0.35, Z = ?3.41, p = 0.001 and 1.75 ± 0.88 to 1.17 ± 1.05, Z = ?4.02, p < 0.001), INL cyst prevalence decreased (index: 0.35 ± 0.52 to 0.28 ± 0.52, Z = ?1.60, p = 0.109), blocking phenomenon did not change significantly (index: 00.12 ± 0.16 to 0.13 ± 0.15, Z = ?0.45, p = 0.656) and subretinal fluid almost disappeared (index: 0.10 ± 0.24 vs. 0.00 ± 0.01, Z = ?2.56, p = 0.011). Correlation analyses revealed highest significant correlations between ONL cyst prevalence and their size and CRT as well as BCVA and MP before treatment and over the course of treatment. Conclusions:  ONL cysts and their size as morphological parameters correlate with retinal function measured with BCVA and microperimetry before and over the course of anti‐VEGF therapy with Ranibizumab in patients with DME.     

Design and evaluation of a chitosan–aprotinin conjugate for the peroral delivery of therapeutic peptides and proteins susceptible to enzymatic degradation

One main barrier for the peroral administration of therapeutic peptides and proteins is the enzymatic barrier, that is mediated by luminally secreted and membrane bound proteolytic enzymes. It was the aim of the study to synthesise, characterise and evaluate a novel polymer–inhibitor conjugate in order to improve the bioavailability of orally-administered peptides and proteins. The trypsin/chymotrypsin inhibitor aprotinin was covalently bound to chitosan. The percentage of the inhibitor in the polymer–inhibitor conjugate (m/m) was determined to be between 1.11 ± 0.36 and 1.92 ± 0.05%. In vitro enzyme assays clearly demonstrated the potential of the novel conjugate to inhibit trypsin and chymotrypsin. Moreover, studies in rats were performed to evaluate the efficacy of the conjugate in vivo. Eight hours after oral administration of tablets containing insulin and the novel chitosan–aprotinin conjugate, the mean blood glucose level decreased to 84 ± 6%. In contrast, the mean blood glucose level in the control group increased to 121 ± 8% of the initial measured blood glucose level. In conclusion it was demonstrated that chitosan–aprotinin conjugate represents a novel and promising tool for the oral administration of therapeutic peptides and proteins susceptible to enzymatic degradation caused by trypsin and chymotrypsin.     

A ribavirin-induced ORF2 single-nucleotide variant produces defective hepatitis E virus particles with immune decoy function

Hepatitis E virus (HEV) is the causative agent of hepatitis E in humans and is the leading cause of enterically transmitted viral hepatitis worldwide. Ribavirin (RBV) is currently the only treatment option for many patients; however, cases of treatment failures or posttreatment relapses have been frequently reported. RBV therapy was shown to be associated with an increase in HEV genome heterogeneity and the emergence of distinct HEV variants. In this study, we analyzed the impact of eight patient-derived open reading frame 2 (ORF2) single-nucleotide variants (SNVs), which occurred under RBV treatment, on the replication cycle and pathogenesis of HEV. The parental HEV strain and seven ORF2 variants showed comparable levels of RNA replication in human hepatoma cells and primary human hepatocytes. However, a P79S ORF2 variant demonstrated reduced RNA copy numbers released in the supernatant and an impairment in the production of infectious particles. Biophysical and biochemical characterization revealed that this SNV caused defective, smaller HEV particles with a loss of infectiousness. Furthermore, the P79S variant displayed an altered subcellular distribution of the ORF2 protein and was able to interfere with antibody-mediated neutralization of HEV in a competition assay. In conclusion, an SNV in the HEV ORF2 could be identified that resulted in altered virus particles that were noninfectious in vitro and in vivo, but could potentially serve as immune decoys. These findings provide insights in understanding the biology of circulating HEV variants and may guide development of personalized antiviral strategies in the future.

Despite its rising global prevalence, hepatitis E is a disease that is mostly overlooked. Every year, more than 44,000 people die as a result of ∼20 million infections worldwide (1). Healthy individuals usually display no or only mild symptoms of viral hepatitis, such as fever, nausea, vomiting, and abdominal pain (2), while patients with preexisting liver disease, pregnant women, and immunocompromised individuals suffer from liver cirrhosis and liver failure (3). Pregnant women additionally present with increased mortality rates of >25% (4). Despite those liver-associated problems, there are also extrahepatic manifestations, such as hematopoietic disease, neurological disorders, and renal injury (5–9). The underlying agent, hepatitis E virus (HEV), is classed within the species of Paslahevepirus balayani (10), formerly known as Orthohepevirus A, which includes isolates from human, swine, wild boar, rat, and other mammals. HEV is a quasienveloped virus existing as both enveloped and non-enveloped particles (11, 12). To date, eight distinct genotypes (GT) of this species of the single-stranded RNA virus have been described (13), which display similar genomic structures. The positive orientated HEV genome is organized in three main open reading frames (ORF1 to ORF3) with a total length of 7.2 kb. Nonstructural proteins forming the HEV replicase complex, such as the RNA-dependent RNA polymerase (RdRp), RNA helicase, or methyltransferase, are encoded by ORF1, while the viral capsid protein is encoded by ORF2. During the HEV replication cycle, HEV produces at least three forms of ORF2 protein: infectious ORF2 (ORF2i), glycosylated ORF2 (ORF2g), and cleaved ORF2 (ORF2c) protein (14). The ORF2i protein is the structural component of infectious particles that is likely derived from the assembly of the intracellular ORF2 (ORF2intra) protein form. In contrast, ORF2g and ORF2c protein are not associated with infectious virions, but secreted in large amounts and are the most abundant antigens detected in patient sera (14). ORF3 encodes for a functional ion channel required for assembly and release of infectious particles by interacting with a variety of host factors (15).In immunocompetent patients, acute hepatitis E usually does not involve antiviral therapy; however, chronically infected and immunocompromised patients often require clinical intervention to clear the infection. Antiviral therapies include pegylated interferon (16–18), successfully implemented for many virus infections, and sofosbuvir (19, 20), a direct acting antiviral against hepatitis C virus, both of which have not yet been systematically evaluated in the context of HEV therapy. Recent studies have investigated the antiviral potential of silvestrol (21), zinc salts (22), and other possible drug candidates in vitro [reviewed in detail by Kinast et al. (23)], but the findings remain to be clinically validated. Lacking specific treatment options, the broad antiviral ribavirin (RBV) (24) is frequently used off-label. However, RBV therapy is often discontinued due to adverse side effects and is only effective in ∼80% of patients, implying that 20% of treated patients remain viremic (25). RBV treatment is specifically contraindicated in pregnant women and can give rise to variants such as G1634R, as well as other amino acid substitutions within the ORF1-encoded polyprotein, potentially contributing to treatment failure and poor clinical long-term outcomes (26–28). In this context, we recently identified viral populations of HEV harboring variations in the capsid-encoding ORF2 region during RBV therapy. With the use of an efficient HEV cell-culture model system, we characterized the impact of these ORF2 variants in the HEV replication cycle.     

Limited Effects of SARS-CoV-2 Pandemic-related Lockdowns and Reduced Population Mobility on Preterm Birth Rates: A Secondary Analysis of Bavarian Obstetric Quality Parameters from 2010 to 2020

Introduction International studies on preterm birth rates during COVID-19 lockdowns report different results. This study examines preterm birth rates during lockdown periods and the impact of the mobility changes of the population in Bavaria, Germany. Material and Methods This is a secondary analysis of centrally collected data on preterm births in Bavaria from 2010 to 2020. Preterm births (< 37 weeks) in singleton and twin pregnancies during two lockdowns were compared with corresponding periods in 2010 – 2019. Fisherʼs exact test was used to compare raw prevalence between groups. Potential effects of two fixed lockdown periods and of variable changes in population mobility on preterm birth rates in 2020 were examined using additive logistic regression models, adjusting for long-term and seasonal trends. Results Unadjusted preterm birth rates in 2020 were significantly lower for singleton pregnancies during the two lockdown periods (Lockdown 1: 5.71% vs. 6.41%; OR 0.88; p < 0.001; Lockdown 2: 5.71% vs. 6.60%; OR = 0.86; p < 0.001). However, these effects could not be confirmed after adjusting for long-term trends (Lockdown 1: adj. OR = 0.99; p = 0.73; Lockdown 2: adj. OR = 0.96; p = 0.24). For twin pregnancies, differences during lockdown were less marked (Lockdown 1: 52.99% vs. 56.26%; OR = 0.88; p = 0.15; Lockdown 2: 58.06% vs. 58.91%; OR = 0.97; p = 0.70). Reduced population mobility had no significant impact on preterm birth rates in singleton pregnancies (p = 0.14) but did have an impact on twin pregnancies (p = 0.02). Conclusions Reduced preterm birth rates during both lockdown periods in 2020 were observed for singleton and twin pregnancies. However, these effects are reduced when adjusting for long-term and seasonal trends. Reduced population mobility was associated with lower preterm birth rates in twin pregnancies. Key words: COVID-19, SARS-CoV-2, preterm birth, lockdown measures, preterm delivery     

Update Breast Cancer 2022 Part 5 – Early Stage Breast Cancer

The treatment of patients with early stage breast cancer has changed in recent years due to the introduction of pembrolizumab, olaparib, and abemaciclib. These and other drugs with the same class of active ingredient are currently in trial for various indications. This review article summarizes the latest results that have either been presented at major conferences such as the ESMO 2022 or published recently in international journals. This includes reports on newly discovered breast cancer genes, atezolizumab in neoadjuvant therapy in HER2-positive patients, long-term data from the APHINITY study, and on how preoperative peritumoral application of local anesthetics can influence the prognosis. We also present solid data on dynamic Ki-67 from the ADAPT studies.Key words: breast cancer, surgery, chemotherapy, therapy standard     

Synthesis of Zeolites from Fine-Grained Perlite and Their Application as Sorbents

The hydrothermal alteration of perlite into zeolites was studied using a two-step approach. Firstly, perlite powder was transformed into Na-P1 (GIS) or hydro(xy)sodalite (SOD) zeolites at 100 °C and 24 h using 2 or 5 M NaOH solutions. Secondly, the Si:Al molar ratio of the reacted Si-rich solution was adjusted to 1 by Na-aluminate addition to produce zeolite A (LTA) at 65 or 95 °C and 6 or 24 h at an efficiency of 90 ± 9% for Al and 93 ± 6% for Si conversion. The performance of these zeolites for metal ion removal and water softening applications was assessed by sorption experiments using an artificial waste solution containing 4 mmol/L of metal ions (Me²⁺: Ca²⁺, Mg²⁺, Ba²⁺ and Zn²⁺) and local tap water (2.1 mmol/L Ca²⁺ and 0.6 mmol/L Mg²⁺) at 25 °C. The removal capacity of the LTA-zeolite ranged from 2.69 to 2.86 mmol/g for Me²⁺ (=240–275 mg/g), which is similar to commercial zeolite A (2.73 mmol/g) and GIS-zeolite (2.69 mmol/g), and significantly higher compared to the perlite powder (0.56 mmol/g) and SOD-zeolite (0.88 mmol/g). The best-performing LTA-zeolite removed 99.8% Ca²⁺ and 93.4% Mg²⁺ from tap water. Our results demonstrate the applicability of the LTA-zeolites from perlite for water treatment and softening applications.     

Influence of fluoride concentration and ethanol pre-treatment on the reduction of the acid susceptibility of enamel

Objective

To determine the association between KOH-soluble and structurally bound fluoride uptake and the erosion resistance of enamel, respectively. Additionally, the effect of enamel pre-treatment with ethanol before fluoridation was assessed.

Methods

Sixty bovine incisors (4 specimens/tooth) were randomly allocated to six groups (A-F). Samples 1 and 2 remained untreated, serving as control at baseline. Pre-treatment of the samples was performed for 5 min with 99% ethanol (groups A, B and C) or physiologic saline (groups D, E and F). Samples 3 and 4 were treated either with 0.5% (groups A and D), 1.0% (groups B and E) or 1.5% (groups C and F) fluoride solution. In samples 1 and 3, uptake of KOH-soluble and structurally bound fluoride was determined. Samples 2 and 4 were used for the determination of acid susceptibility by immersion in 1 ml HCl for 30 s. Calcium release into HCl was assessed by atomic absorption spectroscopy. Differences between the groups were calculated by unpaired t-tests (p < 0.05).

Results

Mode of pre-treatment showed no influence on fluoride acquisition. KOH-soluble and structurally fluoride uptake increased with increasing fluoride concentrations. Highest acid resistance was observed after treatment with 1% fluoride solution for both kinds of pre-treatment followed by 1.5% and 0.5% fluoride solution.

Conclusion

Dose-dependency was observed for enamel fluoride acquisition but not for acid resistance.