Increasing hepatitis B vaccination coverage and decreasing hepatitis B co-infection prevalence among people with HIV-1 in Germany, 1996-2019. Results from a cohort study primarily in men who have sex with men.

OBJECTIVES: Viral hepatitis co-infection among people living with HIV is known to accelerate the progression of liver disease and AIDS. An increased prevalence and incidence of hepatitis B virus (HBV) infection among people living with HIV demands continuous monitoring to adapt targeted prevention strategies to reach the global goals of eliminating viral hepatitis as a public health threat. METHODS: We determined the prevalence and incidence of HBV for the years 1996-2019 from yearly blood sample testing and questionnaire reports among people living with HIV belonging to a nationwide, multicentre observational, prospective cohort study. RESULTS: Among this study population of 3479 participants, the majority (87%) indicated that being men who have sex with men (MSM) was their likely HIV transmission route; 51% were recruited from Berlin. HBV prevalence for acute/chronic and resolved infections decreased from 4.1% and 45% in 1996-1999 to 1.3% and 16% in 2019, respectively. Simultaneously, participants with a serological status indicating HBV vaccination increased from 25% in 1996-1999 to 69% in 2019. Among vaccinated participants with relevant information (n = 1135), 38% received their first HBV vaccination after HIV infection. The HBV incidence rate in 565 eligible participants decreased from 6.9/100 person-years in 2004-2007 to 0.45/100 person-years in 2015. CONCLUSION: Increasing vaccination coverage because of a general HBV vaccination recommendation and catch-up vaccination efforts among risk groups decreased HBV infection prevalence over time among this study population of people living with HIV, primarily MSM and from Berlin. Despite this success, the prevalence and incidence of HBV remains higher than in the general population in Germany. This emphasizes the need for continued HBV prevention by promoting HBV vaccination and HBV screening at regular intervals based on the individual risk behaviour.

Serological survey of Bartonella spp., Borrelia burgdorferi, Brucella spp., Coxiella burnetii, Francisella tularensis, Leptospira spp., Echinococcus, Hanta-, TBE- and XMR-virus infection in employees of two forestry enterprises in North Rhine-Westphalia, Germany, 2011-2013.

We initiated a survey to collect basic data on the frequency and regional distribution of various zoonoses in 722 employees of forestry enterprises in the German state of North Rhine-Westphalia (NRW) from 2011 to 2013. Exposures associated with seropositivity were identified to give insight into the possible risk factors for infection with each pathogen. 41.2% of participants were found to be seropositive for anti-Bartonella IgG, 30.6% for anti-Borrelia burgdorferi IgG, 14.2% for anti-Leptospira IgG, 6.5% for anti-Coxiella burnetii IgG, 6.0% for anti-Hantavirus IgG, 4.0% for anti-Francisella tularensis IgG, 3.4% for anti-TBE-virus IgG, 1.7% for anti-Echinococcus IgG, 0.0% for anti-Brucella IgG and anti-XMRV IgG. Participants seropositive for B. burgdorferi were 3.96 times more likely to be professional forestry workers (univariable analysis: OR 3.96; 95% CI 2.60-6.04; p<0.001); and participants seropositive for Hantavirus 3.72 times more likely (univariable analysis: OR 3.72; 95% CI 1.44-9.57; p=0.007). This study found a surprisingly high percentage of participants seropositive for anti-B. henselae IgG and for anti-F. tularensis IgG. The relatively high seroprevalence for anti-Leptospira IgG seen in this study could be related to living conditions rather than to exposure at work. No specific risk for exposure to C. burnetii and Echinococcus was identified, indicating that neither forestry workers nor office workers represent a risk population and that NRW is not a typical endemic area. Forestry workers appear to have higher risk for contact with B. burgdorferi-infected ticks and a regionally diverse risk for acquiring Hantavirus-infection. The regional epidemiology of zoonoses is without question of great importance for public health. Knowledge of the regional risk factors facilitates the development of efficient prevention strategies and the implementation of such prevention measures in a sustainable manner.

Silicon nitride grids are compatible with correlative negative staining electron microscopy and tip-enhanced Raman spectroscopy for use in the detection of micro-organisms.

AIMS: Successive application of negative staining transmission electron microscopy (TEM) and tip-enhanced Raman spectroscopy (TERS) is a new correlative approach that could be used to rapidly and specifically detect and identify single pathogens including bioterrorism-relevant viruses in complex samples. Our objective is to evaluate the TERS-compatibility of commonly used electron microscopy (EM) grids (sample supports), chemicals and negative staining techniques and, if required, to devise appropriate alternatives. METHODS AND RESULTS: While phosphortungstic acid (PTA) is suitable as a heavy metal stain, uranyl acetate, paraformaldehyde in HEPES buffer and alcian blue are unsuitable due to their relatively high Raman scattering. Moreover, the low thermal stability of the carbon-coated pioloform film on copper grids (pioloform grids) negates their utilization. The silicon in the cantilever of the silver-coated atomic force microscope tip used to record TERS spectra suggested that Si-based grids might be employed as alternatives. From all evaluated Si-based TEM grids, the silicon nitride (SiN) grid was found to be best suited, with almost no background Raman signals in the relevant spectral range, a low surface roughness and good particle adhesion properties that could be further improved by glow discharge. CONCLUSIONS: Charged SiN grids have excellent particle adhesion properties. The use of these grids in combination with PTA for contrast in the TEM is suitable for subsequent analysis by TERS. SIGNIFICANCE AND IMPACT OF THE STUDY: The study reports fundamental modifications and optimizations of the negative staining EM method that allows a combination with near-field Raman spectroscopy to acquire a spectroscopic signature from nanoscale biological structures. This should facilitate a more precise diagnosis of single viral particles and other micro-organisms previously localized and visualized in the TEM.

Sialylated O-glycans and sulfated tyrosines in the NH2-terminal domain of CC chemokine receptor 5 contribute to high affinity binding of chemokines.

The chemokine receptor CCR5 plays an important role in leukocyte chemotaxis and activation, and also acts as a coreceptor for human and simian immunodeficiency viruses (HIV-1, HIV-2, and SIV). We provide evidence that CCR5 is O-glycosylated on serine 6 in the NH2 terminus. The O-linked glycans, particularly sialic acid moieties, significantly contribute to binding of the chemokine ligands. By contrast, removal of O-linked oligosaccharide exerted little effect on HIV-1 infection. Sulfation of specific tyrosine residues in the CCR5 NH2 terminus was important for efficient beta-chemokine binding. Thus, as has been observed for the binding of selectins and their ligands, O-linked carbohydrates and tyrosine sulfates play major roles in promoting the interaction of chemokines with CCR5. The resulting flexible arrays of negative charges on the CCR5 surface may allow specific, high-affinity interactions with diverse chemokine ligands. Although this is the first example of O-linked oligosaccharides and tyrosine sulfates playing a role in chemokine binding, the high density of serines, threonines and tyrosines in the N-termini of many CC chemokine receptors suggests that these posttranslational modifications may commonly contribute to chemokine binding.

Detection limit of negative staining electron microscopy for the diagnosis of bioterrorism-related micro-organisms.

AIMS: To determine the detection limit of diagnostic negative staining electron microscopy for the diagnosis of pathogens that could be used for bioterrorism. METHODS AND RESULTS: Suspensions of vaccinia poxvirus and endospores of Bacillus subtilis were used at defined concentrations as a model for poxviruses and spores of anthrax (Bacillus anthracis), both of which are pathogens that could be used for bioterrorist attacks. Negative staining electron microscopy was performed directly or after sedimentation of these suspensions on to the sample supports using airfuge ultracentrifugation. For both virus and spores, the detection limit using direct adsorption of a 10-µl sample volume onto the sample support was 10(6) particles per ml. Using airfuge ultracentrifugation with a sample volume of 80 µl, the detection limit could be reduced to 10(5) particles per ml for spores and to 5 × 10(4) particles per ml for poxviruses. The influence on particle detection of incubation time, washing and adsorption procedures was investigated. CONCLUSIONS: The reproducibility and sensitivity of the method were acceptable, particularly considering the small sample volume and low particle number applied onto the sample support. SIGNIFICANCE AND IMPACT OF THE STUDY: Diagnostic negative staining electron microscopy is used for the diagnosis of pathogens in emergency situations because it allows a rapid examination of all particulate matter down to the nanometre scale. This study provides precise detection limit for the method, an important factor for the validation and improvement of the technique.

bdhA-patD operon as a virulence determinant, revealed by a novel large-scale approach for identification of Legionella pneumophila mutants defective for amoeba infection.

Legionella pneumophila, the causative agent of Legionnaires' disease, is an intracellular parasite of eukaryotic cells. In the environment, it colonizes amoebae. After being inhaled into the human lung, the bacteria infect and damage alveolar cells in a way that is mechanistically similar to the amoeba infection. Several L. pneumophila traits, among those the Dot/Icm type IVB protein secretion machinery, are essential for exploiting host cells. In our search for novel Legionella virulence factors, we developed an agar plate assay, designated the scatter screen, which allowed screening for mutants deficient in infecting Acanthamoeba castellanii amoebae. Likewise, an L. pneumophila clone bank consisting of 23,000 transposon mutants was investigated here, and 19 different established Legionella virulence genes, for example, dot/icm genes, were identified. Importantly, 70 novel virulence-associated genes were found. One of those is L. pneumophila bdhA, coding for a protein with homology to established 3-hydroxybutyrate dehydrogenases involved in poly-3-hydroxybutyrate metabolism. Our study revealed that bdhA is cotranscribed with patD, encoding a patatin-like protein of L. pneumophila showing phospholipase A and lysophospholipase A activities. In addition to strongly reduced lipolytic activities and increased poly-3-hydroxybutyrate levels, the L. pneumophila bdhA-patD mutant showed a severe replication defect in amoebae and U937 macrophages. Our data suggest that the operon is involved in poly-3-hydroxybutyrate utilization and phospholipolysis and show that the bdhA-patD operon is a virulence determinant of L. pneumophila. In summary, the screen for amoeba-sensitive Legionella clones efficiently isolated mutants that do not grow in amoebae and, in the case of the bdhA-patD mutant, also human cells.

A large proportion of people who inject drugs are susceptible to hepatitis B: Results from a bio-behavioural study in eight German cities.

BACKGROUND: People who inject drugs (PWID) are at high risk of hepatitis B virus (HBV) infection by sharing needles and drug use paraphernalia. In Germany, no routine surveillance of HBV prevalence and vaccination coverage among PWID exists. METHODS: Socio-demographic and behavioural data were collected between 2011 and 2014 through face-to-face interviews, during a bio-behavioural survey of PWID recruited in eight German cities. Dried blood spots (DBS) prepared with capillary blood were tested for HBV markers. Factors associated with past/current HBV infection and vaccination status were analysed by univariable and multivariable analysis using logistic regression. The validity of self-reported HBV infection and vaccination status was analysed by comparison to the laboratory results. RESULTS: Among 2077 participants, the prevalence of current HBV infection was 1.1%, of past HBV infection was 24%, and of vaccine-induced HBV antibodies was 32%. No detectable HBV antibodies were found in 43%. HBV infection status was significantly associated with study city, age, years of injecting, use of stimulants, migration status, and homelessness; HBV vaccination status was significantly associated with study city, age, and level of education. Correct infection status was reported by 71% and correct vaccination status by 45%. CONCLUSIONS: HBV seroprevalence among PWID was about five times higher than in the general population in Germany, confirming PWID as an important risk group. Targeted information campaigns on HBV and HBV prevention for PWID and professionals in contact with PWID need to be intensified. Routinely offered HBV vaccination during imprisonment and opioid substitution therapy would likely improve vaccination rates among PWID.

[Actual food intake of small children at a child day care center]. / Untersuchungen zur realen Nahrungsaufnahme von Kleinkindern in einer Kinderkrippe.

Investigations on the real food intake of children in day-care centres in the G.D.R. have not yet been carried out. Former studies compared the food intake with the recommendations for the daily intake of energy and nutrients. We have studied the daily food intake of 80 children in a day-care centre over a period of three weeks using the "simplified weighing method". The food intake at home was studied additionally by using a protocol method. The intake of energy during the stay in the day-care centre (three meals) amounted to 90 percent of the recommended quantity. The proportion of carbohydrates was lower and that of fat higher than the recommendations. The intake of iron and vitamin C was below the recommended doses. The total caloric intake and intake of fat per day were above the recommended values. Improvement of the composition of meals will help to overcome the disproportions observed.

[Ergometric exercise tests in the diagnosis of juvenile hypertension]. / Ergometrische Belastungsuntersuchungen in der Diagnostik der juvenilen Hypertonie.

We analysed 186 exercise stress-tests in children and adolescents with juvenile hypertension. 40 per cent of our patients showed an exaggerated rise of blood pressure during ergometric testing. There were no severe complications during exercise-tests. We recommended an antihypertensive drug therapy in cases of hypertensive resting blood pressure and hypertensive exercise-test. The ergometric investigation can control the effectiveness of drug therapy. Our data documented that 53.5 per cent showed a spontaneous remission of their elevated blood pressure after 3-5 years. There was no statistical dependence on the exaggerated rise of blood pressure during dynamic exercise-test.

[Medium high-dose estrogen/gestagen sequential therapy in constitutional stature. The results of a 10-year pediatric gynecological follow-up observation]. / Mittelhochdosierte Estrogen/Gestagen-Sequentialtherapie bei konstitutionellem Hochwuchs. Ergebnisse einer 10jährigen gynäkologisch-pädiatrischen Nachbeobachtung.

We report about results of growth-inhibiting treatment in Magdeburg between 1976-1986. In 57 girls closed treatment was studied. Side effects are small. The functional state of the hypothalamo-pituitary axis experienced no negative influence.

[Hypertension outpatient clinic for children and adolescents. Analysis of nearly 10 years of monitoring]. / Hypertoniedispensaire für Kinder und Jugendliche. Analyse einer fast 10jährigen Betreuung.

314 children with hypertension living in an urban district have been transmitted to our out patient clinic. These children were discovered by regularly performed examinations of the medical youth care since 1979. We found a secondary cause of hypertension in 2.2 per cent. Risk factors, as for example familiar disposition, hyperlipidaemia, smoking and overweight, were also analysed. A dynamic exercise-test was performed by means of bicycle-ergometry. 43 per cent of the children with elevated blood pressure in rest reacted with hypertension during exercise. Criteria for drug-therapy were the unsuccessful non-medicamentous effects on the hypertension and a hypertension exercise-test. After 2-5 years 55.9 per cent of all children with elevated blood pressure of the beginning of our study became normotensive without drug-therapy.

[Histological findings of the small intestinal mucosa in rotavirus infections in infants and young children]. / Histologische Befunde der Dünndarmschleimhaut bei Rotavirusinfektionen im Säuglings- und Kleinkindalter.

In a prospective study we performed small intestinal mucosal biopsies in 40 children with acute rotavirus diarrhoea. Biopsies were taken from these children between the 2nd to 10th day of acute phase. The children were at an age of less than 18 month. 95% of the patients had normal histological findings of the small intestinal mucosa. Only 2 children (5%) had well defined injuries of intestinal mucosa. Correlations were not found between clinical findings, morphological results and therapy. In one child with gastroenteritis and meningitis rotaviruses were found in cerebrospinal fluid. In 31.25% of all children rotavirus infection was acquired by a nosocomial infection. The treatment of rotavirus infection is symptomatic. Usually intestinal mucosal biopsy as a routine diagnostic method isn't recommended and it should be used only in intractable courses.

[Parameters of mineral metabolism in children and adolescents in athletic training]. / Mineralstoffwechselparameter bei trainierenden Kindern und Jugendlichen.

Changes in bone structure like chondronecrosis and osteonecrosis occurred in a group of adolescent athletes especially gymnasts. To determine the etiology calcium, inorganic phosphorus, alkaline phosphatase activity and levels of 25-OH-Vit D were investigated. Single low calcium values were present in nearly the half of the athletes. In 37% the vitamin-D-levels were depressed. Such results were found in athletes with and without changes in bone structure. A vitamin D supplementation is recommended if low vitamin D levels are found. Vitamin D deficiency is not the only reason for the disturbances of the bone. Other factors are to take in consideration like nutrition, delayed bone maturation and its consequences, and the mechanical load.

The level of CD4 expression limits infection of primary rhesus monkey macrophages by a T-tropic simian immunodeficiency virus and macrophagetropic human immunodeficiency viruses.

The entry of primate immunodeficiency viruses into cells is dependent on the interaction of the viral envelope glycoproteins with receptors, CD4, and specific members of the chemokine receptor family. Although in many cases the tropism of these viruses is explained by the qualitative pattern of coreceptor expression, several instances have been observed where the expression of a coreceptor on the cell surface is not sufficient to allow infection by a virus that successfully utilizes the coreceptor in a different context. For example, both the T-tropic simian immunodeficiency virus (SIV) SIVmac239 and the macrophagetropic (M-tropic) SIVmac316 can utilize CD4 and CCR5 as coreceptors, and both viruses can infect primary T lymphocytes, yet only SIVmac316 can efficiently infect CCR5-expressing primary macrophages from rhesus monkeys. Likewise, M-tropic strains of human immunodeficiency virus type 1 (HIV-1) do not infect primary rhesus monkey macrophages efficiently. Here we show that the basis of this restriction is the low level of CD4 on the surface of these cells. Overexpression of human or rhesus monkey CD4 in primary rhesus monkey macrophages allowed infection by both T-tropic and M-tropic SIV and by primary M-tropic HIV-1. By contrast, CCR5 overexpression did not specifically compensate for the inefficient infection of primary monkey macrophages by T-tropic SIV or M-tropic HIV-1. Apparently, the limited ability of these viruses to utilize a low density of CD4 for target cell entry accounts for the restriction of these viruses in primary rhesus monkey macrophages.

[Differential diagnosis and therapy of hypoglycemia in childhood]. / Differentialdiagnostik und Therapie der Hypoglykämie im Kindesalter.

Occurrence of hypoglycaemia is directly connected to fuel balance. Compared to free fatty acids, glucose is rapidly exhausted. Therefore, besides disturbances in available glucose (i.e. defects in gluconeogenesis or glycogenolysis) as well as their hormonal control, defects of beta-oxidation might be possible causes. According to recent investigations they occurred with similar frequency. A blood glucose level of 2.6 mmol/l (= 47 mg%) already represents a critical concentration of brain function. Based on pathophysiological concepts concentrations of glucose, free fatty acids, ketone bodies and insulin must be determined from a single plasma sample. Together with case history and clinical symptom findings, these parameters are useful to get a reliable tentative diagnosis. Therefore, descriptions like "ketotic" or "idiopathic" hypoglycaemia can be avoided and suitable treatment can be started. Suspected diagnosis must be confirmed by means of chromatographic, enzymatic, and if necessary histological investigations. Successful treatment depends mainly on early diagnosis. This is of importance in case of surgery in organic hyperinsulinism, but likewise for advisory help to families with infants suffering from beta-oxidation defects, to avoid prolonged fasting.

Molecular cloning and sequence analysis of interleukin 16 from nonhuman primates and from the mouse.

Interleukin 16 (IL-16) is synthesized as a 67 000 Mr precursor (pro-IL-16), but only a carboxy terminal part of 12 000-14 000 Mr is secreted by CD8(+) lymphocytes. This lymphokine binds to CD4 and has been shown to induce migration, affect the activation state of T cells, and inhibit immunodeficiency virus replication. It has been suggested that CD8(+) cell-derived soluble factors play a pivotal role in protecting natural-host nonhuman primates from developing immunodeficiency following SIV infection. In a first attempt to address this question, we cloned and sequenced the IL-16 cDNA from different primates. Here we report the pro-IL-16 sequence from chimpanzees, African green monkeys (AGM), rhesus macaques, and cynomolgus macaques. In order to compare and analyze structural motifs possibly involved in processing, intracellular targeting, or secretion, we extended our study to the New World monkeys saimiri and aotus and to the mouse. Alignments of deduced amino acids reveal that the human protein shares 99% similarity to that of chimpanzees, approximately 95% to rhesus, cynomolgus and AGM, about 90% to aotus and saimiri, and 77.5% to the mouse. Phylogenetic analyses revealed the expected evolutionary groupings.

[Necrotizing enterocolitis in the newborn infant. Pathogenetic risk factors in a 3 year analysis]. / Die nekrotisierende Enterokolitis des Neugeborenen. Pathogenetische Risikofaktoren in einer 3-Jahres-Analyse.

In a clinical study possible risk factors causing necrotizing enterocolitis (NEC) were studied in 17 preterm infants with NEC and compared with a control group of 17 healthy preterm infants. We found a more frequent rate of blood exchange transfusions and a lower rate of breast milk feeding in patients who later developed NEC than in the control group. But there were no differences between both groups concerning factors promoting hypoxia of the gut, the number of infections, antibiotic treatment, or hyperosmolar enteral feeding. The preterm infants with NEC did not show any characteristics in history, physical examination and clinical course. These children could not be distinguished as a risk group for this illness from the newborn infant control group. Therefore, with the exception of breast milk feeding, no special prevention of the necrotizing enterocolitis can be recommended.

Molecular cloning, sequence, expression, and processing of the interleukin 16 precursor.

Interleukin 16 (IL-16) has been shown to function as chemoattractant factor, as a modulator of T-cell activation, and as an inhibitor of immunodeficiency virus replication. The recent identification of inconsistencies in published IL-16 cDNA nucleotide sequences led to the proposal that IL-16 is synthesized in the form of a large precursor protein (pro-IL-16). To identify the true transcriptional start of the IL-16 mRNA rapid amplification of cDNA ends methods were applied. The complete pro-IL-16 cDNA was subsequently molecularly cloned, sequenced, and expressed in COS-7 cells. We report here that pro-IL-16 is most likely synthesized as a 67-kDa protein and is encoded from a major 2.6-kb transcript. Recombinant pro-IL-16 polypeptides are specifically cleaved in lysates of CD8(+) cells, suggesting that the naturally secreted bioactive form of IL-16 is smaller than the originally published 130 amino acids fragment. Moreover, in contrast to other interleukins such as IL-15, IL-16 mRNA expression is almost exclusively limited to lymphatic tissues underlining the potential of IL-16 as an immune regulatory molecule.

GA-binding protein factors, in concert with the coactivator CREB binding protein/p300, control the induction of the interleukin 16 promoter in T lymphocytes.

Interleukin 16 (IL-16) is a chemotactic cytokine that binds to the CD4 receptor and affects the activation of T cells and replication of HIV. It is expressed as a large 67-kDa precursor protein (pro-IL-16) in lymphocytes, macrophages, and mast cells, as well as in airway epithelial cells from asthmatics after challenge with allergen. This pro-IL-16 is subsequently processed to the mature cytokine of 13 kDa. To study the expression of IL-16 at the transcriptional level, we cloned the human chromosomal IL-16 gene and analyzed its promoter. The human IL-16 gene consists of seven exons and six introns. The 5' sequences up to nucleotide -120 of the human and murine IL-16 genes share >84% sequence homology and harbor promoter elements for constitutive and inducible transcription in T cells. Although both promoters lack any TATA box, they contain two CAAT box-like motifs and three binding sites of GA-binding protein (GABP) transcription factors. Two of these motifs are part of a highly conserved and inducible dyad symmetry element shown previously to control a remote IL-2 enhancer and the CD18 promoter. In concert with the coactivator CREB binding protein/p300, which interacts with GABPalpha, the binding of GABPalpha and -beta to the dyad symmetry element controls the induction of IL-16 promoter in T cells. Supplementing the data on the processing of pro-IL-16, our results indicate the complexity of IL-16 expression, which is tightly controlled at the transcriptional and posttranslational levels in T lymphocytes.