Adapter CAR T cells to counteract T-cell exhaustion and enable flexible targeting in AML.

Although the landscape for treating acute myeloid leukemia (AML) patients has changed substantially in recent years, the majority of patients will eventually relapse and succumb to their disease. Allogeneic stem cell transplantation provides the best anti-AML treatment strategy, but is only suitable in a minority of patients. In contrast to B-cell neoplasias, chimeric antigen receptor (CAR) T-cell therapy in AML has encountered challenges in target antigen heterogeneity, safety, and T-cell dysfunction. We established a Fab-based adapter CAR (AdCAR) T-cell platform with flexibility of targeting and control of AdCAR T-cell activation. Utilizing AML cell lines and a long-term culture assay for primary AML cells, we were able to demonstrate AML-specific cytotoxicity using anti-CD33, anti-CD123, and anti-CLL1 adapter molecules in vitro and in vivo. Notably, we show for the first time the feasibility of sequential application of adapter molecules of different specificity in primary AML co-cultures. Importantly, using the AML platform, we were able to demonstrate that chronic T-cell stimulation and exhaustion can be counteracted through introduction of treatment-free intervals. As T-cell exhaustion and target antigen heterogeneity are well-known causes of resistance, the AdCAR platform might offer effective strategies to ameliorate these limitations.

Comparison of the Effects of Beclomethasone Dipropionate and Budesonide in the Treatment of Children with Mild, Persistent Asthma.

Objective To compare the mean change in peak expiratory flow values in children receiving inhaled beclomethasone dipropionate versus inhaled budesonide in the treatment of mild persistent asthma. Method The medical records of 60 patients from the outpatient department (OPD)/emergency room (ER), National Institute of Child Health, Karachi, who received beclomethasone dipropionate (BDP) 200 µg one puff and budesonide (BUD) 200 µg one puff twice a day for treatment of mild persistent asthma from March 10, 2020, to August 10, 2020, were explored. Results The mean age of children was 10.56 ± 3.01 years in the BUD group and 10.05 ± 3.54 years in the BDP group. The mean change in peak expiratory flow % in the BUD group was 15.69 ± 3.59%, and in the BDP group, it was 13.59 ± 4.26% (P-value=0.04) Conclusion BDP and budesonide (BUD) were both found to be effective for the treatment of mild persistent asthma in children. However, we found that BUD had better efficacy compared to BDP.

Pattern and Outcome of Dengue Fever in a Pediatric Tertiary Hospital: A Retrospective Report.

Introduction For Pakistan, dengue has been established as a public health problem. With superimposed factors such as poor socioeconomic conditions, limited public health awareness, poor hygiene, and sanitation conditions, the situation has become more severe and complications have become frequent. Almost 90% of all infections occur in children of age less than 18 years. This is a three-year retrospective report of dengue fever in Southern Pakistan. Methods In this retrospective analysis, all records of patients admitted to the National Institute of Child Health, Karachi, from May 1, 2016, till April 30, 2019, diagnosed with dengue fever were recruited. Their demographic, clinical, and biochemical records were assessed. The outcome was also recorded. Data were entered and analyzed using Statistical Package for Social Sciences (SPSS) for Windows version 20.0 (IBM Corp., Chicago). Results Among 93 cases of dengue fever, there were 71 (76.3%) male and 22 (23.7%) female children. Their mean age was 5.7 ± 3.07 years. The mean duration from onset of disease to hospitalization was 4.2 ± 2.1 days. The mean platelet count was 47391.30 ± 41370.61 x 109/L. Fever (100%) and abdominal pain (35.5%) were common presentations. Bleeding episodes were seen in 31% of children, rash in 15%, disseminated intravascular coagulation in 3%, and 1% developed pleural effusion. There were no mortalities; 87 (93.5%) were discharged and six (6.5%) children left against medical advice. Conclusion Fever, abdominal pain, bleeding episodes, and rash were common presentations. Hematological, hepatological, neurological, and pleural complications were not uncommon. The outcome of the disease was adequate and there were no mortalities.

Pre- and periconceptional primary cytomegalovirus infection: risk of vertical transmission and congenital disease.

OBJECTIVE: To estimate the risk of congenital cytomegalovirus infection and disease following primary maternal infection around the time of conception compared with the risk during later stages of pregnancy. DESIGN: Cohort study between 1990 and 2003. SETTING: Germany. PARTICIPANTS: One hundred and sixty-six pregnant women with serologically confirmed primary cytomegalovirus infection and known outcome. METHODS: Timing of primary cytomegalovirus infection by analysing the kinetics of cytomegalovirus-specific IgG and IgM antibodies, the IgG avidity index and neutralising antibodies. MAIN OUTCOME MEASURE: Onset of maternal primary infection in relation to congenital infection and disease. RESULTS: Preconceptional (between eight and two weeks before onset of the last menstrual period) was determined in three women and did not lead to congenital infection. Periconceptional infection (between one week before and five weeks after last menstrual period) occurred in 20 women with congenital infection in nine cases (45%). Timing was less precise (between eight weeks before and five weeks after last menstrual period) in an additional 10 women, three cases of which resulted in congenital infection. Of the 12 pregnancies in which congenital infection occurred, seven were terminated, six before the 12th week of gestation (WG 12) and one at WG 19 due to fetal hyperechogenic bowel. One of the five infected live-born infants delivered to a mother with periconceptional infection showed dystrophy and mild microcephaly at birth, but had a rather normal development at two years of age. Primary infections occurring between WG 6-20 and WG 20-38 resulted in transmission rates of 30% (27/89) and 58% (18/31), respectively. CONCLUSIONS: Counselling of women with periconceptional primary cytomegalovirus infection should be adjusted to offer prenatal diagnosis and high-level ultrasound controls due to the considerable risk for fetal infection and uncertainty of clinical outcome and disease.

[Androgenetic alopecia in the man]. / Die androgenetische Alopezie des Mannes.

Androgenetic alopecia (AGA) occurs in approximately 40% of men at the age of 40 and 50% at 50, respectively. Especially for young men progressive hair loss can be distressing. Therefore, understanding of these patients' concerns is important for appropriate management. Current understanding of the pathophysiology of AGA mainly focuses on androgen metabolism as it affects hair growth. As a result, pharmacologic treatment has made considerable progress through the introduction of selective 5 alpha-reductase inhibition with finasteride. In placebo-controlled clinical trials in men with AGA, treatment with oral finasteride proved to be effective. Minoxidil is the only pharmacological substance for topical application with proven efficacy. So far, other treatment modalities have no proven efficacy in clinical trials, so that their use cannot be recommended. Options for advanced AGA not amenable to pharmacologic treatment are autologous hair transplantation and hair replacement, both of which have recently also made progress in terms of cosmetic appeal.

[Primary headache in childhood and adolescence. The association between changes in family interaction patterns and therapy success--a pilot study]. / Primäre Kopfschmerzen im Kindes- und Jugendalter.

INTRODUCTION: Family interaction patterns are often involved in diseases and disorders in childhood and adolescence in complex ways (e.g in their development, maintenance and cure). The present study deals with the role of family factors in success in a pediatric headache therapy consisting of group hypnotherapy and systemic family consultation. METHODS: A sample of 12 outpatients, aged 9-15 years and balanced in sex, is investigated. Patients were diagnosed by IHS-criteria. Global symptom strain was measured by numeric rating scale (NRS) at pre-appointment and at 9-months follow-up appointment. Also family interaction patterns associated with the occurrence of headache symptoms were measured by content analysis. RESULTS: We found an association between changes in two independently assessed variables: global symptom strain and family interaction patterns. (1) When patients assessed global symptom strain as unchanged, family interaction patterns associated with headache were also assessed as unchanged by observers; (2) when patients assessed their global symptom strain as positively changed, family interaction pattern associated with headache were also assessed as positively changed by observers. CONCLUSION: These data provide empirical evidence about when to include family in treatment of pediatric headache: when rigid family interaction patterns associated with headache complicate a symptom change.

Prenatal diagnosis of congenital cytomegalovirus infection in 189 pregnancies with known outcome.

Prenatal diagnosis (PD) of fetal cytomegalovirus (CMV) infection was performed in 242 pregnancies, with known outcome in 189 cases. In 141/189 pregnancies, PD was carried out on account of suspicious maternal CMV serology up to gestational week (WG) 23, and in 48 cases on account of abnormal ultrasonic findings detected between WG 18 and 39. Chorionic villus samples (n = 6), amniotic fluid (AF, n = 176) and/or fetal blood specimens (n = 80) were investigated for detection of virus by cell culture, shell vial assay, PCR and/or CMV-specific IgM antibodies. Of 189 fetuses correctly evaluated by CMV detection either in fetal tissue following therapeutic abortion/stillbirth (n = 24) or in urine of neonates within the first 2 weeks of life (n = 33), 57 were congenitally infected. In women with proven or suspected primary infection, the intrauterine transmission rates were 20.6% (7/34) and 24.4% (10/41), respectively. Of the congenitally infected live-born infants, 57.6% (19/33) had symptoms of varying degree. The overall sensitivity of PD in the serologic and ultrasound risk groups was 89.5% (51/57). A sensitivity of 100% was achieved by combining detection of CMV-DNA and CMV-specific IgM in fetal blood or by combined testing of AF and fetal blood for CMV-DNA or IgM antibodies. There was no instance of intrauterine death following the invasive procedure. The predictive value of PD for fetal infection was 95.7% (132/138) for negative results and 100% (51/51) for positive results. Correct results for congenital CMV infection by testing AF samples can be expected with samples obtained after WG 21 and after a time interval of at least 6 weeks between first diagnosis of maternal infection and PD. In case of negative findings in AF or fetal blood and the absence of ultrasound abnormalities at WG 22-23, fetal infection and neonatal disease could be excluded with high confidence. Positive findings for CMV infection in AF and/or fetal blood in combination with CMV suspicious ultrasound abnormalities predicted a high risk of cytomegalic inclusion disease (CID). Furthermore, detection of specific IgM antibodies in fetal blood was significantly correlated with severe outcome for the fetus or the newborn (p = 0.0224). However, normal ultrasound of infected fetuses at WG 22-23 can neither completely exclude an abnormal ultrasound at a later WG and the birth of a severely damaged child nor the birth of neonates which are afflicted by single manifestations at birth or later and of the kind which are not detectable by currently available ultrasonographic techniques.

Selective specimen preparation for TEM observation of the cross-section of individual carbon nanotube/metal junctions

We present here an efficient method to prepare a transmission electron microscopy (TEM) specimen for selective observation of the cross-section of individual nanoscale structures. As a typical example, the cross-sectional TEM observation of a quasi-one-dimensional material - a nano-electronic component based on an individual carbon nanotube - is presented.

Leg ulcers in patients with myeloproliferative disorders: disease- or treatment-related?

Leg ulcers are a relatively frequent problem in patients with myeloproliferative disorders under treatment with hydroxyurea (HU). The pathogenesis is currently unknown and may be multifactorial. Concomitant arterial or venous disease may play a contributing role in the development of these wounds. Vasculitis, cryoglobulinemia and pyoderma gangrenosum should be considered if typical clinical signs are present. We report on 3 patients with myeloproliferative disorders who developed HU-induced leg ulcers and review the literature. HU-induced leg ulcers share clinical features which can help to differentiate them from leg ulcers of other etiologies: occurrence under long-term treatment with HU at a dose of at least 1 g/day, localization in the malleolar region and spontaneous healing when HU is discontinued. We conclude that differentiation between disease-related and treatment-induced leg ulcers can be difficult and may not always be possible. In HU-induced leg ulcers, cessation of the drug typically leads to wound healing.

Combination of microneutralization and avidity assays: improved diagnosis of recent primary human cytomegalovirus infection in single serum sample of second trimester pregnancy.

Estimation of IgG avidity index is a classical serological method. Antibodies with low avidity are detectable at a very early stage of infection whereas high avidity antibodies indicate past infection. Recently, it was shown that the neutralization assay can be routinely used as a reliable method for differentiating between acute primary and non-primary infection in a single serum sample because the first neutralizing titers (NT) appeared after an average of 13 weeks (range, 10-17 weeks). A low positive NT titer in the presence of specific IgM antibodies, however, still represents a diagnostic problem especially if blood sampling occurred after the 12th week of gestation. To overcome this problem the combination of NT and IgG avidity tests was evaluated. Human cytomegalovirus (HCMV) IgG avidity indices of 350 serum samples from 227 pregnant women were investigated using 6M urea in the washing buffer. HCMV specific IgG antibodies reached full maturation approximately 20-22 weeks after seroconversion and low IgG avidity is therefore a marker of primary infection. The combined application of the microneutralization and avidity assays was shown to serve as a helpful tool in diagnosis of a recent primary HCMV infection of second trimester pregnancy particularly when previous serological data were not available.

Evaluation of two novel enzyme immunoassays using recombinant antigens to detect cytomegalovirus-specific immunoglobulin M in sera from pregnant women.

BACKGROUND: Two novel enzyme linked immunosorbent assays (ELISA) (Abbott IMx CMV IgM 2.0, and Cobas Core CMV IgM EIA recomb, research version) which use recombinant antigens to detect cytomegalovirus (CMV)-specific IgM antibodies were evaluated. OBJECTIVES: A new ELISA is normally evaluated against a gold standard commercial kit, which in this case does not exist. We therefore evaluated the two novel recombinant ELISA against four conventional ELISAs and a recently developed CMV IgM immunoblot containing four purified viral and four recombinant proteins. STUDY DESIGN: A total of 280 sera from pregnant women and 42 potentially cross-reactive sera were investigated using the six ELISAs, including 101 sera which were also tested using the new IgM immunoblot. RESULTS: Relative sensitivity, relative specificity and overall agreement differed according to the reference assay. The Cobas Core CMV EIA recomb showed much higher agreement with the ELISA consensus, and the IMx CMV IgM 2.0 with the immunoblot. CONCLUSION: The evaluation of these new IgM assays in terms of their agreement with either commercial ELISA kits or the IgM immunoblot demonstrates that the question 'which reference method?' is still open. However the recombinant IgM assays may improve the diagnosis of CMV infection in pregnancy since the recombinant technology offers helpful tools for identifying diagnostically relevant proteins and allows the use of standardized pure preparations of antigens. For serological diagnosis of CMV infection in pregnancy two IgM assays that can be relied upon should be performed. IgM positive sera should be tested with supplementary assays to differentiate primary from non-primary infection.

[Erythroderma and alcohol abuse]. / Erythrodermie und Athylabusus.

We report on 7 patients with chronic alcohol abuse and ichthyosiform erythroderma. Dry skin (i.e. xerosis cutis) was a striking common feature in all patients who did not present any signs of fluid depletion. Two patients had exclusively exsiccational dermatitis; three had atopic disposition and one nummular eczema of the legs. In another patient a drug allergy could be demonstrated. Xerosis cutis seems to be an important precipitating aetiologic factor for erythroderma, and may be a consequence of alcohol-induced diuresis. All patients recovered rapidly under topical treatment with emollients and low potency steroids. We propose that ichthyosiform erythroderma be added to the known skin diseases caused by chronic alcohol abuse.

Nanosecond optical parametric oscillator based on large-aperture periodically poled RbTiOAsO(4).

We report on optical parametric oscillators (OPO's) based on periodically poled RbTiOAsO(4) (PP RTA), which are pumped by Q -switched solid-state lasers. With a diode-pumped Nd:YVO(4) laser (pulse energy, 800microJ ; pulse duration, 5.5 ns; repetition rate, 1 kHz) the PP RTA OPO generated 1.58-microm signal and 3.26-microm idler radiation with a signal pulse energy of 45microJ . The large aperture of 3 mmx3 mm of the PP RTA crystal also permitted OPO operation with pump pulse energies as high as 65 mJ, provided by a flash-lamp-pumped Q -switched Nd:YAG laser (pulse duration, 20 ns; repetition rate, 10 Hz). With this pump source the OPO generated signal pulse energies as high as 17 mJ, corresponding to an efficiency of 26%. The performance of this OPO shows that large-aperture PP RTA crystals are well suited for pulsed nanosecond OPO operation with pump pulse energies of tens of millijoules.

Nanosecond periodically poled lithium niobate optical parametric generator pumped at 532 nm by a single-frequency passively Q-switched Nd:YAG laser.

We report an efficient, visible, nanosecond optical parametric generator of periodically poled lithium niobate pumped at 532 nm by a frequency-doubled, diode-pumped, passively Q -switched, single-mode Nd:YAG laser with 90-muJ pulse energy. The signal radiation is tunable from 637 to 593 nm. The maximum signal-conversion efficiency is 23%. Optical parametric amplification of a He-Ne laser at 632.8 nm is also studied.

Bilateral deafness due to congenital CMV and not rubella reinfection.

We report on an infant with bilateral deafness discovered at the age of 5 months caused by a retrospectively diagnosed primary maternal CMV infection after definitive exclusion of maternal rubella reinfection as a cause of fetal infection.

An enzyme linked immunoassay using recombinant antigens for differentiation of primary from secondary or past CMV infections in pregnancy.

BACKGROUND: As primary cytomegalovirus (CMV) infection in pregnancy may be associated with severe fetal outcome, the serological differentiation of primary infection from recurrent or previous infection is of major importance. OBJECTIVES: This differentiation was attempted with a CMV IgG enzyme immunoassay (EIA), which investigated the differential IgG immune response to recombinant proteins p52 and pp150. To express the IgG reactivity to either protein a ratio was calculated by OD p52/OD pp150. STUDY DESIGN: Serial samples from groups of pregnant women with primary infection (n = 18) were compared to those with recurrent (n = 7) or previous CMV infection (n = 189). RESULTS: In primary infected women a predominant IgG response to p52 (p52 alone or ratio > or = 1.5) was observed in early sera less than 4 weeks after seroconversion, whereas the IgG response to recombinant protein pp150 was delayed and appeared after 2-7 weeks. Women with secondary and those with past infection had either IgG antibodies to pp150 alone or a ratio of less than 1.5 in 85 and 89.1% respectively with no remarkable change of ratio over time. CONCLUSIONS: The IgG recombinant EIA was shown to be a useful supplementary assay for differentiation of primary up to 8 weeks after seroconversion from recurrent or past infections.

The action of benzbromarone in relation to age, sex and accompanying diseases.

The uric-acid lowering effect was investigated in a group of 2220 patients. 1984 of these were employed for statistical evaluation purposes. On average, the uric acid level was reduced from 8,24 +/- 1,16 mg/100 ml to 5,32 +/- 1,265 mg/100 ml. In 82 % of all the cases, the uric acid level at the end of the treatment period was below 6,4 mg%, both in patients treated throughout with 50 mg benzbromarone (NarcaricinRmite) and in those changed over to 100 mg benzbromarone (NarcaricinR) after 1 to 3 weeks. The lowering of the uric acid levels was in no way related to hypertension, adiposity, hyperlipoproteinaemia, diabetes mellitus or age.