Dutch Type Research Articles

Residual C-peptide is associated with new and persistent impaired awareness of hypoglycaemia in type 1 diabetes

AimsTo describe the change in impaired awareness of hypoglycaemia (IAH) over time and to identify factors associated with this change in the Dutch Type 1 Diabetes biomarkers cohort (NCT04977635). MethodsA prospective cohort of type 1 diabetes patients, with C-peptide <300 pmol/L, who had completed the Clarke questionnaire, to determine IAH status, at baseline and after 2 years. Changes in awareness status were defined and compares as follows: unchanged normal awareness (NAH) versus unchanged IAH, new IAH versus reversal of IAH. Multivariate logistic regression models were fitted using forward and backward stepwise selection using a 0.10 P-value cut-off, and stepwise backward selection using AIC criteria. ResultsA total of 431 out of 611 participants were included. The baseline prevalence of IAH was 17 % and 20 % after 2 years. The incidence proportion of new IAH and reversal of IAH were, 9.5 % and 31 %, respectively. For every 2.7-fold increase in C-peptide, the odds of IAH decrease by 58 %. A 1-unit increase in BMI over the 2-year follow-up period is associated with a 5.27-fold increase in the odds of reversing IAH. ConclusionsHigher C-peptide levels are protective against new IAH, and an increase in BMI over time is associated with the reversal of IAH.

Metabolomic associations of impaired awareness of hypoglycaemia in type 1 diabetes

This study investigates impaired awareness of hypoglycaemia (IAH), a complication of insulin therapy affecting 20–40% of individuals with type 1 diabetes. The exact pathophysiology is unclear, therefore we sought to identify metabolic signatures in IAH to elucidate potential pathophysiological pathways. Plasma samples from 578 individuals of the Dutch type 1 diabetes biomarker cohort, 67 with IAH and 108 without IAH (NAH) were analysed using the targeted metabolomics Biocrates AbsoluteIDQ p180 assay. Eleven metabolites were significantly associated with IAH. Genome-wide association studies of these 11 metabolites identified significant single nucleotide polymorphisms (SNPs) in C22:1-OH and phosphatidylcholine diacyl C36:6. After adjusting for the SNPs, 11 sphingomyelins and phosphatidylcholines were significantly higher in the IAH group in comparison to NAH. These metabolites are important components of the cell membrane and have been implicated to play a role in cell signalling in diabetes. These findings demonstrate the potential role of phosphatidylcholine and sphingomyelins in IAH.

Minocycline for sporadic and hereditary cerebral amyloid angiopathy (BATMAN): study protocol for a placebo-controlled randomized double-blind trial

BackgroundCerebral amyloid angiopathy (CAA) is a disease caused by the accumulation of the amyloid-beta protein and is a major cause of intracerebral hemorrhage (ICH) and vascular dementia in the elderly. The presence of the amyloid-beta protein in the vessel wall may induce a chronic state of cerebral inflammation by activating astrocytes, microglia, and pro-inflammatory substances. Minocycline, an antibiotic of the tetracycline family, is known to modulate inflammation, gelatinase activity, and angiogenesis. These processes are suggested to be key mechanisms in CAA pathology. Our aim is to show the target engagement of minocycline and investigate in a double-blind placebo-controlled randomized clinical trial whether treatment with minocycline for 3 months can decrease markers of neuroinflammation and of the gelatinase pathway in cerebrospinal fluid (CSF) in CAA patients.MethodsThe BATMAN study population consists of 60 persons: 30 persons with hereditary Dutch type CAA (D-CAA) and 30 persons with sporadic CAA. They will be randomized for either placebo or minocycline (15 sporadic CAA/15 D-CAA minocycline, 15 sporadic CAA/15 D-CAA placebo). At t = 0 and t = 3 months, we will collect CSF and blood samples, perform a 7-T MRI, and collect demographic characteristics.DiscussionThe results of this proof-of-principle study will be used to assess the potential of target engagement of minocycline for CAA. Therefore, our primary outcome measures are markers of neuroinflammation (IL-6, MCP-1, and IBA-1) and of the gelatinase pathway (MMP2/9 and VEGF) in CSF. Secondly, we will look at the progression of hemorrhagic markers on 7-T MRI before and after treatment and investigate serum biomarkers.Trial registrationClinicalTrials.gov NCT05680389. Registered on January 11, 2023

Reproductive performance of cows of different ecotypes of black-and-white breeds

Abstract It was studied the main indicators of the functionality of reproductive system and their interrelation with the productivity indices of imported cows populations Holstein breed, belonging to 3 ecological types: the Dutch, the French and the German and our indigenous Moldovian type Black-Motley breed. The service period, was the shortest in cows of Moldovian type of Black-Motley breed with 38.1 days (P˂0.001), 30.2 days (P˂0.001) and by 14.5 days (P˂0.01.), compared to the Holstein German, Dutch and French ecological types, respectively. The difference of the duration of calvings interval between indigenous population and French ecological type was 18.2 days (p&lt;0.01), compared to the German 46 days (P&lt;0.001) and to the Dutch 36,5 days (P&lt;0.001). There has been a difference of 9,9% in birth coefficient, between Moldovian type and Holstein German ecological type (p˂0.05). The best result of the coefficient of reproductive capacity has the indigenous type, opposite of German ecological type, which has the lowest performance (P˂0.001). Between the service period and the productivity indicators of the milk, regardless of the origin of the animal populations, were predominantly established, authentic positive correlative interrelationships, from weak to strong and statistically negative between productivity indices and the coefficient of the reproductive capacity of cows.

Referral rate, profile and degree of control of patients with familial hypercholesterolemia: data from a single lipid unit from a Mediterranean area

BackgroundThe challenging rigorous management of hypercholesterolemia promotes referral to specialized units. This study explored the need, based on referral rate and cardiovascular (CV) risk factor control in patients evaluated for familial hypercholesterolemia (FH), for a lipid unit (LU).MethodsOver a four-year period, 340 referrals to our unit were analyzed to establish the lipid disorder referral rate. Moreover, 118 patients referred for potential FH during the period 2010–2018 (52.4 ± 13.9 years, 47.5% male, Caucasian, 26.3% obese, 33.1% smokers and 51.7% with some glycaemic alteration) were investigated. The Dutch Lipid Clinic Network (DLCN) score, type and dose of lipid-lowering drugs, lipid profile including lipoprotein (a) (Lp(a)) and the presence of plaques with carotid ultrasound (CU) were recorded.ResultsLipids represented 6.2% of referrals (38 patient-years) requiring a 2–3 h weekly monographic outpatient consultation. The potential FH sample displayed a DLCN score ≥ 6 in 78% and modifiable CV risk factors in 51%. Only 22% achieved tight disease control despite intensive treatment. The statin-ezetimibe combination treatment group achieved better goals (73.0% vs. 45.5%, P = 0.003), and the rosuvastatin group had a higher proportion of prediabetes (60.9% vs. 39.1%, P = 0.037). Neither CU plaque presence nor Lp(a) > 50 mg/dL was linked with established CV disease patients, but higher Lp(a) concentrations were detected between them (102.5 (26.3–145.8) vs. 25.0 (13.0–52.0) mg/dL, P = 0.012).ConclusionsThe referral rate, degree of control, and proportion of modifiable CV risk factors in FH patients demonstrate the need for LU in our area as well as optimize control and treatment.

Annals of Neurology: Volume 92, Number 3, September 2022

Spatial patterns of white matter injury in Dutch type Cerebral Amyloid Angiopathy (D‐CAA). A: FLAIR‐based white matter hyperintensity probability map in D‐CAA carriers showing a posterior predominance. Individual images were placed in Montreal Neurological Institute (MNI) space, and voxels are colored by the probability of a white matter lesion being present in this cohort. B: Diffusion based mean diffusivity (MD) in the white matter skeleton. Individual MD images were placed in MNI space, and voxels are colored by the average value across D‐CAA carriers in this cohort. See paper in this issue by Shirzadi et al. (pp. 358–363).

Whole-genome sequence analysis of high-level penicillin-resistant strains and antimicrobial susceptibility of Neisseria gonorrhoeae clinical isolates from Thailand.

The increasing rate of antimicrobial-resistant Neisseria gonorrhoeae poses a considerable public health threat due to the difficulty in treating gonococcal infections. This study examined antimicrobial resistance (AMR) to drugs recommended for gonorrhea treatment between 2015 and 2017, and the AMR determinants and genetic compositions of plasmids in 3 gonococcal strains with high-level penicillin resistance. We collected 117 N. gonorrhoeae isolates from patients with gonococcal infections who attended Siriraj Hospital, Bangkok, Thailand, between 2015 and 2017. Minimum inhibitory concentrations (MICs) of penicillin, tetracycline, ciprofloxacin, azithromycin, spectinomycin, cefixime, and ceftriaxone were determined by the agar dilution method. PCR amplification and sequencing of 23S rRNA and mtrR (a negative regulator of MtrCDE efflux pump) were performed. Whole genomes of 3 PPNG strains with high-level penicillin resistance (MIC ≥ 128 μg/ml) were sequenced using Illumina and Nanopore sequencing platforms. The proportions of N. gonorrhoeae isolates with resistance were 84.6% for penicillin, 91.5% for tetracycline, and 96.6% for ciprofloxacin. All isolates were susceptible to spectinomycin, azithromycin, cefixime, and ceftriaxone. An adenine deletion within a 13 bp inverted repeat sequence in the mtrR promoter and an H105Y mutation in the mtrR coding region were found in the N. gonorrhoeae isolate with the highest azithromycin MIC value (1 μg/ml). Three high-level penicillin-resistant isolates contained nonmosaic type II penA and had mutations in penB and the mtrR coding region. All isolates with high-level penicillin resistance carried the conjugative plasmids with or without the Dutch type tetM determinant, the beta-lactamase plasmid (Rio/Toronto), and the cryptic plasmid. The gonococcal population in Thailand showed high susceptibility to ceftriaxone and azithromycin, current dual therapy recommended for gonorrhea treatment. As elevated MIC of azithromycin has been observed in 1 strain of N. gonorrhoeae, expanded and enhanced surveillance of antimicrobial susceptibility and study of genetic resistance determinants are essential to improve treatment guidelines.

Identifying and delineating the type 2 diabetes population in the Netherlands using an all-payer claims database: characteristics, healthcare utilisation and expenditures

ObjectivesWe aimed to identify and delineate the Dutch type 2 diabetes population and the distribution of healthcare utilisation and expenditures across the health system from 2016 to 2018 using an...

Alignment between outcomes and minimal clinically important differences in the Dutch type 2 diabetes mellitus guideline and healthcare professionals' preferences.

To evaluate the clinical benefit of new medicines for type 2 diabetes mellitus (T2DM), the Dutch guideline committee T2DM in primary care established the importance of outcomes and minimal clinically important differences (MCIDs). The present study used an online questionnaire to investigate healthcare professionals’ opinions about the importance of outcomes and preferences for MCIDs. A total of 211 physicians, pharmacists, practice nurses, diabetes nurses, nurse practitioners and physician assistants evaluated the importance of mortality, macro‐ and microvascular morbidity, HbA1c, body weight, quality of life, (overall) hospital admissions and severe and other hypoglycemia on a 9‐point scale. All outcomes were considered critical (mean scores 7–9), except for body weight and other hypoglycemia (mean scores 4–6). Only HbA1c and hospital admissions were valued differently by the guideline committee (not critical). Other relevant outcomes according to the respondents were adverse events, ease of use and costs. Median MCIDs were 4 mmol/mol for HbA1c (guideline: 5 mmol/mol) and 3 kg for body weight (guideline: 5 kg weight gain and 2,5 kg weight loss). Healthcare professionals preferred relative risk reductions of 20% for mortality (guideline: 10%) and macrovascular morbidity (guideline: 25%) and 50% for other hypoglycaemia (guideline: 25%). The MCID of 25% for microvascular morbidity, hospital admissions and severe hypoglycaemia corresponded to the guideline‐MCID. Healthcare professionals’ preferences were thus comparable to the views of the guideline committee. However, healthcare professionals had a stricter view on the importance of HbA1c and hospital admissions and the MCIDs for mortality and other hypoglycemia.

Development of a salutogenic intervention for healthy eating among Dutch type 2 diabetes mellitus patients.

Lay SummaryHealthy eating can be challenging for type 2 diabetes mellitus (T2DM) patients. The theory of salutogenesis, which focuses on the resources required to organize behavioural changes in everyday life, was used to develop an intervention for healthy eating. The aim was to describe the development, structure and content of this salutogenic intervention. The development consisted of two phases that were based on the operationalization of important key principles of salutogenesis. In Phase 1 (Exploration and synthesis), a systematic review and three qualitative studies were performed to explore important characteristics to enable healthy eating in everyday life. The results were used to develop the draft intervention. In Phase 2 (Validation and adjustment), interviews and workshops were conducted with T2DM patients, healthcare providers and scientists. Based on this, the draft intervention was modified into its final form. The developmental process resulted in a 12-week, group-based intervention that aimed to enable important resources for healthy eating via self-examination, reflection, setting goals and sharing experiences. Attention was also paid to disease information, disease acceptance, food literacy, stress management, self-identity and social support. The group sessions began following an individual intake session, with a booster session held 3 months after the intervention. The researcher’s translation of the stakeholders’ priorities into an intervention was corrected for and approved by the stakeholders concerned. This comprehensive salutogenic intervention was developed based on practical and scientific evidence. Providing transparency in developmental processes and content is important because it determines the scientific integrity and credibility of an intervention.

Cerebral amyloid angiopathy is associated with decreased functional brain connectivity

Cerebral amyloid angiopathy (CAA) is a major cause of intracerebral hemorrhage and neurological decline in the elderly. CAA results in focal brain lesions, but the influence on global brain functioning needs further investigation. Here we study functional brain connectivity in patients with Dutch type hereditary CAA using resting state functional MRI. Twenty-four DNA-proven Dutch CAA mutation carriers (11 presymptomatic, 13 symptomatic) and 29 age-matched control subjects were included. Using a set of standardized networks covering the entire cortex, we assessed both within- and between-network functional connectivity. We investigated group differences using general linear models corrected for age, sex and gray matter volume. First, all mutation carriers were contrasted against control subjects and subsequently presymptomatic- and symptomatic mutation carriers against control subjects separately, to assess in which stage of the disease differences could be found. All mutation carriers grouped together showed decreased connectivity in the medial and lateral visual networks, default mode network, executive control and bilateral frontoparietal networks. Symptomatic carriers showed diminished connectivity in all but one network, and between the left and right frontoparietal networks. Presymptomatic carriers also showed diminished connectivity, but only in the frontoparietal left network. In conclusion, global brain functioning is diminished in patients with CAA, predominantly in symptomatic CAA and can therefore be considered to be a late consequence of the disease.

Biological microchip for assessing tetracycline-resistance in &lt;i&gt;Neisseria gonorrhoeae&lt;/i&gt; clinical isolates in Russian Federation

A total of 399 Neisseria gonorrhoeae clinical isolates collected in different regions of the Russian Federation in 20152017 were analyzed for tetracycline susceptibility and genetic markers of resistance. Drug susceptibility testing was performed by serial dilution method in agar and minimum inhibitory concentration (MIC) was measured according to the Russian “Guidelines for microbial susceptibility testing for antibacterial agents No. 4.2.1890-04”. Tetracycline resistance determinants were studied by using hydrogel microarray with immobilized oligonucleotide probes able to identify a series of chromosomal mutations and detect plasmid tetM gene. Different resistance determinants were found in 193 isolates (48.4%). Mutation in codon 57 in the rpsJ gene (41.2%) was most common that decreases tetracycline affinity to ribosome 30S subunit, mainly due to Val57Met substitution both as a point mutation as well as in combination with others. Mutations in the rpsJ gene were found in strains with the intermediate tetracycline susceptibility. Mutations in the porB gene (lower tetracycline influx) held the se cond place in prevalence pattern (23.1%); the Gly120Lys substitution usually led to emergence of tetracycline resistance either as a point mutation or in combination with other substitutions. Substitutions of Gly120 for other residues (Asp, Asn, and Thr) and Ala121 for Asp, Asn, and Gly had much less effect on resistance level. The –35 delA deletion in the promoter region of mtrR gene (increased expression of MtrC-MtrD-MtrE efflux pump) was observed in 11.3% strains. The tetM gene was found in 27 strains including 17 American and 10 Dutch type tetM determinants. Evolutionary tree was constructed for the tetM genes with the estimation of their homology with similar genes in genera Streptococcus, Enterococcus and Mycoplasma. Mutations in chromosomal genes resulted in increase of tetracycline MIC up to 2–4 mg/L; 4 mg/L MIC was observed in case of simultaneous presence of several mutations. Strains bearing tetM gene-containing plasmid showed extremely high resistance level: MIC ≥ 8 mg/L (64 mg/L for the two samples). Thus, long-lasting withdrawal of tetracycline use for treatment of gonococcal infections in Russia (since 2003) resulted in decreased percentage of resistant strains (including strains with intermediate susceptibility) from 75% down to 45.4%. However, currently tetracycline resistance in Russia remains elevated that is explained by the presence of different resistance determinants in the half of isolates under study.

Mapping Cucumber Vein Yellowing Virus Resistance in Cucumber (Cucumis sativus L.) by Using BSA-seq Analysis.

Cucumber vein yellowing virus (CVYV) causes severe yield losses in cucurbit crops across Mediterranean countries. The control of this virus is based on cultural practices to prevent the presence of its vector (Bemisia tabaci) and breeding for natural resistance, which requires the identification of the loci involved and the development of molecular markers for linkage analysis. In this work, we mapped a monogenic locus for resistance to CVYV in cucumber by using a Bulked Segregant Analysis (BSA) strategy coupled with whole-genome resequencing. We phenotyped 135 F3 families from a segregating population between a susceptible pickling cucumber and a resistant Long Dutch type cucumber for CVYV resistance. Phenotypic analysis determined the monogenic and incomplete dominance inheritance of the resistance. We named the locus CsCvy-1. For mapping this locus, 15 resistant and 15 susceptible homozygous F2 individuals were selected for whole genome resequencing. By using a customized bioinformatics pipeline, we identified a unique region in chromosome 5 associated to resistance to CVYV, explaining more than 80% of the variability. The resequencing data provided us with additional SNP markers to decrease the interval of CsCvy-1 to 625 kb, containing 24 annotated genes. Markers flanking CsCvy-1 in a 5.3 cM interval were developed for marker-assisted selection (MAS) in breeding programs and will be useful for the identification of the target gene in future studies.

Secernin-1 is a novel phosphorylated tau binding protein that accumulates in Alzheimer\u2019s disease and not in other tauopathies

We recently identified Secernin-1 (SCRN1) as a novel amyloid plaque associated protein using localized proteomics. Immunohistochemistry studies confirmed that SCRN1 was present in plaque-associated dystrophic neurites and also revealed distinct and abundant co-localization with neurofibrillary tangles (NFTs). Little is known about the physiological function of SCRN1 and its role in Alzheimer’s disease (AD) and other neurodegenerative diseases has not been studied. Therefore, we performed a comprehensive study of SCRN1 distribution in neurodegenerative diseases. Immunohistochemistry was used to map SCRN1 accumulation throughout the progression of AD in a cohort of 58 patients with a range of NFT pathology (Abundant NFT, n = 21; Moderate NFT, n = 22; Low/No NFT, n = 15), who were clinically diagnosed as having AD, mild cognitive impairment or normal cognition. SCRN1 accumulation was also examined in two cases with both Frontotemporal Lobar Degeneration (FTLD)-Tau and AD-related neuropathology, cases of Down Syndrome (DS) with AD (n = 5), one case of hereditary cerebral hemorrhage with amyloidosis – Dutch type (HCHWA-D) and other non-AD tauopathies including: primary age-related tauopathy (PART, [n = 5]), Corticobasal Degeneration (CBD, [n = 5]), Progressive Supranuclear Palsy (PSP, [n = 5]) and Pick’s disease (PiD, [n = 4]). Immunohistochemistry showed that SCRN1 was a neuronal protein that abundantly accumulated in NFTs and plaque-associated dystrophic neurites throughout the progression of AD. Quantification of SCRN1 immunohistochemistry confirmed that SCRN1 preferentially accumulated in NFTs in comparison to surrounding non-tangle containing neurons at both early and late stages of AD. Similar results were observed in DS with AD and PART. However, SCRN1 did not co-localize with phosphorylated tau inclusions in CBD, PSP or PiD. Co-immunoprecipitation revealed that SCRN1 interacted with phosphorylated tau in human AD brain tissue. Together, these results suggest that SCRN1 is uniquely associated with tau pathology in AD, DS and PART. As such, SCRN1 has potential as a novel therapeutic target and could serve as a useful biomarker to distinguish AD from other tauopathies.

The metabolomic approach to the complex biochemical characteristics of cole &lt;i&gt;Brassica oleracea&lt;/i&gt; L.

Relevance A commonly cultivated and demanded type of vegetable crops – cabbage Brassica oleracea L. – in the process of evolution and domestication was divided into three clusters: leafy cabbage, headed cabbage and broccoli (cauliflower). According to modern data, cabbage originated in culture from the large-leaved forms of leafy cabbage. Portuguese tronchuda is a transitional form to a typical headed cabbage. It is represented by leafy and semi-capped forms and evolutionarily related to Portuguese halega (Galician) leafy cabbage. White cabbage (Brassica oleracea L. convar. capitata (DC.) Alef. var. capitata L. f. alba DC.) is the most significant in the world and Russian economy. The characteristic of cultures of Brassica oleracea L. (varieties of white cabbage and tronchuda) is given according to the most important biochemical quality characteristics. Methods The research material included 48 samples of cabbage, including 27 samples of 13 European cultivar types, including Dutch, Central European and South European cultivar types, 8 samples – two Russian northwestern and central Russian cultivar types and 13 samples – five oriental cultivar types of Caucasian and Turkish-Balkan origin, 9 samples – Portuguese cabbage tronchuda from the VIR collection, reflecting the ecological, geographic and genetic diversity of cabbage. Samples of cabbage were grown at the research and production base "Pushkin and Pavlovsk laboratories VIR" (St. Petersburg), biochemical analysis was carried out in the laboratory of biochemistry and molecular biology of VIR. Results As a result of using of the modern non-specific metabolic approach to the study of the biochemical composition in samples of white cabbage, about 100 components have been identified from the groups of organic acids, free amino acids, including essential, fatty acids, including essential, polyhydric alcohols, sugars, and also phenolic compounds, wax, nucleosides, and others. Regularity in the accumulation of nutrient and biologically active substances by cultures of Brassica oleracea L., by cultivar types and individual samples were revealed. As a result of our research using modern techniques, including the nonspecific metabolic approach, new data on the biochemical composition of white and semi-capped forms were obtained. The cabbage within the studied botanical varieties and groups of cultivar types has a complex biochemical composition that characterizes cabbage samples as samples with potentially high value (the role and importance of far from all compounds in the organization of healthy human nutrition is known), which confirms the need for in-depth control of the biochemical composition of plants when breeding new varieties. Samples with the optimal component composition for balanced human nutrition, which are proposed to be used in breeding for quality, including obtaining varieties for healthy (functional) and therapeutic and prophylactic nutrition of the population of theRussian Federation were found.

Early Aβ reduction prevents progression of cerebral amyloid angiopathy.

Clinical trials targeting β-amyloid peptides (Aβ) for Alzheimer disease (AD) failed for arguable reasons that include selecting the wrong stages of AD pathophysiology or Aβ being the wrong target. Targeting Aβ to prevent cerebral amyloid angiopathy (CAA) has not been rigorously followed, although the causal role of Aβ for CAA and related hemorrhages is undisputed. CAA occurs with normal aging and to various degrees in AD, where its impact and treatment is confounded by the presence of parenchymal Aβ deposition. APPDutch mice develop CAA in the absence of parenchymal amyloid, mimicking hereditary cerebral hemorrhage with amyloidosis Dutch type (HCHWA-D). Mice were treated with a β-site amyloid precursor protein cleaving enzyme 1 (BACE1) inhibitor. We used 3-dimensional ultramicroscopy and immunoassays for visualizing CAA and assessing Aβ in cerebrospinal fluid (CSF) and brain. CAA onset in mice was at 22 to 24 months, first in frontal leptomeningeal and superficial cortical vessels followed by vessels penetrating the cortical layers. CSF Aβ increased with aging followed by a decrease of both Aβ40 and Aβ42 upon CAA onset, supporting the idea that combined reduction of CSF Aβ40 and Aβ42 is a specific biomarker for vascular amyloid. BACE1 inhibitor treatment starting at CAA onset and continuing for 4 months revealed a 90% Aβ reduction in CSF and largely prevented CAA progression and associated pathologies. This is the first study showing that Aβ reduction at early disease time points largely prevents CAA in the absence of parenchymal amyloid. Our observation provides a preclinical basis for Aβ-reducing treatments in patients at risk of CAA and in presymptomatic HCHWA-D. ANN NEUROL 2019;86:561-571.

Resistance Profile of Neisseria gonorrhoeae in KwaZulu-Natal, South Africa Questioning the Effect of the Currently Advocated Dual Therapy.

We report on the antimicrobial resistance profile of Neisseria gonorrhoeae isolates and the distribution of tetM genes in isolates with high-level tetracycline resistance in KwaZulu-Natal, South Africa. Male and female patients presenting with urethral and/or vaginal discharge were recruited into the study. Urethral and cervical secretions were cultured on New York City agar. Confirmatory tests for N. gonorrhoeae included Gram stain, catalase, oxidase, and carbohydrate utilization tests. Beta-lactamase was tested by means of the chromogenic cephalosporin test. Minimum inhibitory concentrations were determined using agar dilution with multipoint inoculation. Polymerase chain reaction with gel electrophoresis was used to detect the presence and type of the tetM gene. N. gonorrhoeae was isolated from the specimens of 319 (26%) of the 1220 recruited patients. Of these 319 isolates, 71% were resistant to 3 or more drugs. Resistance to azithromycin was found in 68% of the isolates. All isolates showed high-level tetracycline resistance with minimum inhibitory concentration values of 16 and 32 mg/mL. The tetM gene was present in 293 (92%). The American type was found in 264 (90%) and the Dutch type in 29 (10%). Twenty-six (8%) did not carry a tetM gene. The current syndromic management with dual ceftriaxone and azithromycin is due to the high level of azithromycin resistance factually single-drug therapy. High-level tetracycline resistance based on a resistance mechanism other than ribosome protection by the tetM gene product is present in N. gonorrhoeae infecting South African patients.

Multiple Approaches to Diffusion Magnetic Resonance Imaging in Hereditary Cerebral Amyloid Angiopathy Mutation Carriers.

BackgroundCerebral amyloid angiopathy (CAA) is a major cause of lobar intracerebral hemorrhage in elderly adults; however, presymptomatic diagnosis of CAA is difficult. Hereditary cerebral hemorrhage with amyloidosis–Dutch type (HCHWA‐D) is a rare autosomal‐dominant disease that leads to pathology similar to sporadic CAA. Presymptomatic HCHWA‐D mutation carriers provide a unique opportunity to study CAA‐related changes before any symptoms have occurred. In this study we investigated early CAA‐related alterations in the white matter.Methods and ResultsWe investigated diffusion magnetic resonance imaging (dMRI) data for 15 symptomatic and 11 presymptomatic HCHWA‐D mutation carriers and 30 noncarrier control participants using 4 different approaches. We looked at (1) the relation between age and global dMRI measures for mutation carriers versus controls, (2) voxel‐wise dMRI, (3) independent component‐clustered dMRI measures, and (4) structural connectomics between presymptomatic or symptomatic carriers and controls. Fractional anisotropy decreased, and mean diffusivity and peak width of the skeletonized mean diffusivity increased significantly over age for mutation carriers compared with controls. In addition, voxel‐wise and independent component‐wise fractional anisotropy, and mean diffusivity, and structural connectomics were significantly different between HCHWA‐D patients and control participants, mainly in the periventricular frontal and occipital regions and in the occipital lobe. We found no significant differences between presymptomatic carriers and control participants.ConclusionsThe dMRI technique is sensitive in detecting alterations in symptomatic HCHWA‐d carriers but did not show alterations in presymptomatic carriers. This result indicates that dMRI may be less suitable for identifying early white matter changes in CAA.

Determining the influence of the composition of milk from cows of different breeds on quality indicators for the dutch-type cheese

The paper reports a comprehensive study into the impact of feeding rations and breed of cows on the technological properties of milk and quality characteristics of the Dutch type of cheese. Despite the abundance of information on the relationship between domestic animal feeding and technological parameters for the production of cheeses, a possible relationship between particular properties of animal breeds, changes in feed, and a positive influence on the quality of products requires in-depth research. That explains the relevance of studying the influence of an animal breed and the optimization of animal feed on a set of technological parameters and quality indicators for the Dutch type of cheeses. We manufactured the Dutch-type cheese in a series of experiments using milk from cows of different breeds, obtained at different rations of feeding. We compared the chemical composition of milk from three groups of cows ‒ the Ukrainian brown dairy breed, Lebedinskaya breed, and Simmental breed, at different feed. The results obtained demonstrate that the use of alfalfa haylage contributes to an increase in the mass share of a dry fat-free milk residue in milk, improves its suitability for cheese, as compared with milk obtained over a silage-hay period.We have worked out technological modes for producing the hard Dutch-type cheese from milk of three cow breeds at different feeding rations. It is shown that when applying alfalfa haylage to feed animals the duration of basic technological operations shortens. That helps reduce the share of dry substances (including fat) in whey, and increases the output of finished product. The samples of hard Dutch-type cheese, made from milk of all three groups of animals over a haylage-hay period are characterized by a high content of essential amino acids, as well as free amino acids, and soluble non-protein nitrogen. That predetermined their higher degree of ripeness, and the larger point-based assessment, in terms of organoleptic characteristics (by 7‒15 points), compared to the samples of cheese produced from milk received over a silage-hay period. The highest point-based estimate for organoleptic characteristics (99 points) was given to samples of the Dutch-type cheese made from milk of the Ukrainian brown dairy cow, milked over a haylage-hay period.It was established that an animal breed has a significant impact on the suitability of milk for cheese and yield of the finished product. The most significant influence of the optimized feeding was exerted on the organoleptic and sensorial indicators for a Dutch-type cheese. The proof are the indicators for proteolytic activity. There was an increase in the growth of nitrogen within soluble fractions, an accelerated maturation process, as evidenced by the accumulation of low-molecular nitrogenous fractions. The largest total amount of free amino acids was demonstrated by cheese made from milk of the cows that were fed the improved ration using alfalfa haylage.The developed feeding rations, as well as the improved technological parameters for making a Dutch-type cheese, could be recommended as the cost and product quality management methods for producing mass-market and craft cheese.

Molecular Antimicrobial Resistance of Neisseria gonorrhoeae in a Moroccan Area.

Objectives To identify the prevalence and the types of Neisseria gonorrhoeae (NG) resistance plasmids-mediated penicillin (PPNG) and tetracycline (TRNG), the ciprofloxacin resistance (CRNG), and related risk factors of each types of resistance. Methods The beta-lactamase-producing plasmid types (Africa, Asia, and Toronto), tetM tetracycline resistance plasmid types (America and Dutch), and the determination of the Ser-91 mutation of GyrA were detected by specifics PCRs on 149 diagnosed NG positives samples followed by Hinf1 digestion for tetM and gyrA mutation. Results 135 (90.1%) samples showed a profile of molecular resistance to at least one antibiotic with predominance of ciprofloxacin resistance. In fact, 36 (24.2%) and 69 (46.3%) cases harbored PPNG and TRNG, respectively, and 116 (77.9%) cases showed the mutation Ser-91 of GyrA (CRNG). From a total of 36 PPNG isolates, the Toronto, Asian, and Toronto/Asian types were detected in 13 (36.1%), 10 (27.8%), and 13 (36.1%) cases, respectively, whereas the African type was not detected. In addition, the American type of TRNG was detected in 92.8% (64/69) of cases, while the Dutch type was detected in 7.2% (5/69) of cases. The association of demographics and clinical variables with NG resistance to ciprofloxacin, penicillin, and tetracycline was studied and the risk factors have been determined. Conclusion Resistance to penicillin, tetracycline, and ciprofloxacin among NG samples positives remained at high levels in Morocco as determined by molecular profile. So, the use of molecular tools for NG antimicrobial resistance detection can help in the management and spread limitation of this infection.