Immune Response following BNT162b2 mRNA COVID-19 Vaccination in Pediatric Cancer Patients.

COVID-19 vaccinations are recommended for children with cancer but data on their vaccination response is scarce. This study assesses the antibody and T-cell response following a 2- or 3-dose vaccination with BNT162b2 mRNA COVID-19 vaccine in children (5-17 years) with cancer. For the antibody response, participants with a serum concentration of anti-SARS-CoV-2 spike 1 antibodies of >300 binding antibody units per milliliter were classified as good responders. For the T-cell response, categorization was based on spike S1 specific interferon-gamma release with good responders having >200 milli-international units per milliliter. The patients were categorized as being treated with chemo/immunotherapy for less than 6 weeks (Tx < 6 weeks) or more than 6 weeks (Tx > 6 weeks) before the first immunization event. In 46 patients given a 2-dose vaccination series, the percentage of good antibody and good T-cell responders was 39.3% and 73.7% in patients with Tx < 6 weeks and 94.4% and 100% in patients with Tx > 6 weeks, respectively. An additional 3rd vaccination in 16 patients with Tx < 6 weeks, increased the percentage of good antibody responders to 70% with no change in T-cell response. A 3-dose vaccination series effectively boosted antibody levels and is of value for patients undergoing active cancer treatment.

Molecular analysis of the erythroid phenotype of a patient with BCL11A haploinsufficiency.

The BCL11A gene encodes a transcriptional repressor with essential functions in multiple tissues during human development. Haploinsufficiency for BCL11A causes Dias-Logan syndrome (OMIM 617101), an intellectual developmental disorder with hereditary persistence of fetal hemoglobin (HPFH). Due to the severe phenotype, disease-causing variants in BCL11A occur de novo. We describe a patient with a de novo heterozygous variant, c.1453G>T, in the BCL11A gene, resulting in truncation of the BCL11A-XL protein (p.Glu485X). The truncated protein lacks the 3 C-terminal DNA-binding zinc fingers and the nuclear localization signal, rendering it inactive. The patient displayed high fetal hemoglobin (HbF) levels (12.1-18.7% of total hemoglobin), in contrast to the parents who had HbF levels of 0.3%. We used cultures of patient-derived erythroid progenitors to determine changes in gene expression and chromatin accessibility. In addition, we investigated DNA methylation of the promoters of the γ-globin genes HBG1 and HBG2. HUDEP1 and HUDEP2 cells were used as models for fetal and adult human erythropoiesis, respectively. Similar to HUDEP1 cells, the patient's cells displayed Assay for Transposase-Accessible Chromatin (ATAC) peaks at the HBG1/2 promoters and significant expression of HBG1/2 genes. In contrast, HBG1/2 promoter methylation and genome-wide gene expression profiling were consistent with normal adult erythropoiesis. We conclude that HPFH is the major erythroid phenotype of constitutive BCL11A haploinsufficiency. Given the essential functions of BCL11A in other hematopoietic lineages and the neuronal system, erythroid-specific targeting of the BCL11A gene has been proposed for reactivation of γ-globin expression in ß-hemoglobinopathy patients. Our data strongly support this approach.

Effects of nutritional stress during different developmental periods on song and the hypothalamic-pituitary-adrenal axis in zebra finches.

In songbirds, developmental stress affects song learning and production. Altered hypothalamic-pituitary-adrenal (HPA) axis function resulting in elevated corticosterone (CORT) may contribute to this effect. We examined whether developmental conditions affected the association between adult song and HPA axis function, and whether nutritional stress before and after nutritional independence has distinct effects on song learning and/or vocal performance. Zebra finches (Taeniopygia guttata) were raised in consistently high (HH) or low (LL) food conditions until post-hatch day (PHD) 62, or were switched from high to low conditions (HL) or vice versa (LH) at PHD 34. Song was recorded in adulthood. We assessed the response of CORT to handling during development and to dexamethasone (DEX) and adrenocorticotropic hormone (ACTH) challenges during adulthood. Song learning and vocal performance were not affected by nutritional stress at either developmental stage. Nutritional stress elevated baseline CORT during development. Nutritional stress also increased rate of CORT secretion in birds that experienced stress only in the juvenile phase (HL group). Birds in the LL group had lower CORT levels after injection of ACTH compared to the other groups, however there was no effect of nutritional stress on the response to DEX. Thus, our findings indicate that developmental stress can affect HPA function without concurrently affecting song.

Analysis of steroids in songbird plasma and brain by coupling solid phase extraction to radioimmunoassay.

It is a common practice to extract steroids from plasma, serum, or tissue samples prior to steroid measurement by radioimmunoassay (RIA) or enzyme immunoassay (EIA). Steroid extraction is critical because it can remove substances that interfere with the RIA or EIA. Steroid extraction is commonly achieved using organic solvents, such as diethyl ether or dichloromethane. However, organic solvent extractions can suffer from low recovery, imprecise recovery, or incomplete removal of assay interference. Here, we describe validations of a simple protocol to extract steroids (e.g., dehydroepiandrosterone, corticosterone, and estradiol) from avian plasma, serum, and brain tissue using solid phase extraction (SPE) with commercially available C18 columns. We compare various methods for (1) eluting steroids from columns, (2) drying eluates, and (3) resuspending dried eluates prior to RIA. The SPE method yields high and consistent recoveries. The SPE method also effectively separates steroids from interfering substances, even when extracting steroids from lipid-rich plasma and brain tissue. These data indicate that SPE is superior to organic solvent extraction on several measures. SPE should be broadly useful for extracting steroids from plasma or tissue samples.

Balneotherapy in rheumatic diseases--an overview of novel and known aspects.

Balneotherapeutic applications to treat rheumatic diseases have a long-term tradition and are based on established scientific principles, and the majority of rheumatic diseases most frequently include balneotherapy. However, as other therapeutic strategies, balneological interventions require scientific proof of their effect and efficacy. Notably, recent studies providing evidence of the thermic and chemical effects on the immune system and cytokine milieu have enforced the revival of the traditional balneological interventions. This review provides an overview of the known as well as the novel aspects of balneotherapy and their clinical relevance in the treatment of rheumatic diseases.

Association of 1.25 vitamin D3 deficiency, disease activity and low bone mass in ankylosing spondylitis.

Vertebral fractures due to osteoporosis are a common but frequently unrecognized complication in established ankylosing spondylitis (AS). It is known that inflammatory activity in rheumatic diseases (i.e., proinflammatory cytokines) itself plays a possible role in the pathophysiology of bone loss. The aim of this study was to analyze whether inflammatory activity and an alteration of the vitamin D metabolism play a substantial role in the loss of bone mass in AS. In this cross-sectional study, 58 patients with established AS and an age- and sex-matched control group were examined. The vitamin D status was investigated, as was, in parallel, the relationship to disease activity (erythrocyte sedimentation rate [ESR], C-reactive protein [CRP], Bath Ankylosing Spondylitis Disease Activity Index [BASDAI]), markers of bone metabolism (parathyroid hormone [PTH], 1.25 vitamin D3, 25 vitamin D3), calcium, bone alkaline phosphatase (bone-AP), urine cross-links, and plasma tumor necrosis factor alpha (TNFalpha). Bone mineral density was measured by quantitative computed tomography (QCT) of the lumbar spine. Osteoporosis was diagnosed in early as well as in progressive stages of AS (23/58=39.6%). Furthermore, serum levels of 1.25 vitamin D3 and PTH were negatively correlated with disease activity and TNFalpha. The excretion of cross-links showed a positive correlation with disease activity and TNFalpha, and 1.25 vitamin D3 and PTH were positively correlated with bone-AP. TNFalpha also positively correlated with disease activity. AS patients with osteoporosis showed significantly increased CRP, ESR, cross-links and PTH and a significantly decreased 1.25 D3. Osteoporosis is frequent in AS and high disease activity is associated with an alteration in vitamin D metabolites and increased levels of bone resorption in active AS. Our findings propose a close association of BMD, bone metabolism and inflammatory activity, possibly related to vitamin D inflammation interactions.

Follow-up of ovarian function post-chemotherapy following ovarian cryopreservation and transplantation.

BACKGROUND: The purpose of this study was to assess the ovarian function after treatment of a malignant disease in women who previously had cortical tissue from an entire ovary cryopreserved prior to chemotherapy, and to assess ovarian function after autotransplantation of cryopreserved ovarian tissue. All were treated with chemotherapeutic drugs with an estimated high risk of inducing ovarian failure. METHODS: Twenty-two women with breast cancer (n = 8), Hodgkin's disease (n = 6), non-Hodgkin's (n = 2), leukaemia (n = 5) or brain tumour (n = 1) underwent a clinical examination >18 months after cryopreservation. Three patients with premature ovarian failure had ovarian tissue autotransplanted orthotopically and heterotopically. Ovarian function was assessed by ultrasonography of the remaining ovary and hormone measurements. RESULTS: Nine of 22 women (41%) had sonographic and hormonal signs of ovarian failure with ovarian volumes <1.3 cm3, no antral follicles and high FSH levels (median 57.1 IU/l). Thirteen of the 22 women (59%) still menstruated and 10 had a seemingly normal ovarian function, with a median ovarian volume of 6.8 cm3, a median number of antral follicles of six, FSH <15 IU/l and normal estradiol levels. All three patients with autotransplanted ovarian tissue regained ovarian function as confirmed by return of menses, follicles on ultrasonography and normalized hormone levels. Two embryos were created from the crypreserved tissue after IVF. CONCLUSIONS: Treatment with bone-marrow transplantation and/or high doses of alkylating agents led to ovarian failure in all patients. Autotransplantation of ovarian tissue led to return of ovarian function.

Anti-Müllerian hormone initiates growth of human primordial follicles in vitro.

Survival and growth of follicles in human ovarian tissue is presently only performed with limited success. We evaluated the effect of anti-Müllerian hormone (AMH) and/or testosterone on follicular growth during a 4-week culture period using ovarian cortical tissue from six women in their reproductive years. The cortex of each biopsy was isolated and immediately cryopreserved upon collection and stored in liquid nitrogen. After thawing the tissue was placed in culture. After the culture period all follicles were counted on histological sections and classified for viability and stage of development. Based on evaluation of 6603 follicles it was found that the number of growing follicles significantly increased during the culture period as compared to the uncultured control, irrespective of the composition of the culture medium. Furthermore, significantly more follicles advanced to the primary and secondary stage (p<0.05) in tissue cultured with AMH (54%) as compared to tissue cultured in control medium (41%). The mean diameter of follicles classified as primary follicles was significantly enhanced in tissue cultured in the presence of AMH (p=0.002) and AMH plus testosterone (p<0.001) as compared to that observed in tissue cultured with control medium and medium containing testosterone alone. In contrast the mean diameter of the oocyte and its nucleus remained similar irrespective of culture medium. In conclusion, AMH seems to affect early stages of human follicular development by enhancing recruitment, survival and/or growth during a 4-week culture period.

A new approach to studying angiogenesis in rheumatoid arthritis by means of power Doppler ultrasonography and measurement of serum vascular endothelial growth factor.

OBJECTIVE: To evaluate angiogenesis as an essential component of pannus formation and cartilage destruction in rheumatoid arthritis (RA) using power Doppler ultrasonography (PDUS) and serum vascular endothelial growth factor (VEGF) measurement. METHODS: Twenty-one RA patients with a painful and swollen wrist and 12 healthy controls were examined with ultrasound. By means of standard scans, vascularity near and inside the joint capsule was visualized with PDUS. Two trained investigators performed sonography. Representative video clips were stored and read by two independent investigators, under blinded conditions, with regard to the microvascular Doppler flow being either inside or outside the joint capsule and with respect to a qualitative estimate of the intensity of blood flow, according to a grading from 1 to 3. Serum levels of VEGF were measured with a standard quantitative sandwich ELISA. RESULTS: The power Doppler mode identified increased synovial microvascular blood flow inside the joint capsule in 17 of 21 RA patients (81%) vs one of the healthy controls. We found large variation in serum VEGF levels in RA patients and in healthy controls. The degree of synovial vascularity determined by PDUS showed no correlation with the immediate serum VEGF level in the same patient. CONCLUSION: The high correlation between intra-articular microvascular power Doppler flow and clinical synovitis in RA patients (P<0.0001) indicates that PDUS may be helpful in studying the role of synovial blood vessels in rheumatoid inflammation.

Neutrophil CD177 (NB1 gp, HNA-2a) expression is increased in severe bacterial infections and polycythaemia vera.

The NB1 glycoprotein (CD177, HNA-2a antigen) is exclusively expressed on human neutrophils. As the clinical significance of CD177 expression is unknown, we investigated its expression in healthy individuals before and after stimulation with granulocyte colony-stimulating factor (G-CSF), in patients with rheumatoid arthritis, viral hepatitis, severe bacterial infections and polycythaemia vera. Expression was quantitatively determined by flow cytometry and by real time polymerase chain reaction. Only G-CSF-stimulated individuals and patients with severe bacterial infections and polycythaemia showed a significantly (P < 0.001) increased CD177 expression compared with healthy individuals, indicating that neutrophil CD177 expression can increase significantly in certain clinical conditions.

Embryo quality in natural versus stimulated IVF cycles.

BACKGROUND: The impact of controlled ovarian stimulation (COS) on oocyte and subsequent embryo quality remains controversial. In the present study we have compared embryo quality in natural and stimulated cycles in the same group of patients. METHODS: This retrospective study was comprised of patients with a regular menstrual cycle who had IVF after COS using rFSH in a long GnRH agonist protocol. In all stimulated cycles the patients had fresh embryos transferred and surplus good quality embryos cryopreserved. Subsequently the same patients were treated with a modified FER cycle (mFER) where thawing of the frozen embryos was combined with aspiration of the dominant follicle in the natural cycle. The embryo cleavage stage and quality score were compared between the stimulated and the natural cycle for the patients having an embryo in the natural cycle. RESULTS: In 177 cases patients returned for mFER in a natural cycle. Spontaneous ovulation had occurred in 35 cycles. In 17 cycles no oocyte was retrieved at aspiration and in 125 cycles 128 oocytes were aspirated. In the stimulated cycles from these patients we had obtained 950 embryos (cleavage rate 70.4%) versus 85 embryos (cleavage rate 66.4%) (P = 0.34) in the natural cycles. Comparing the embryos in the natural and stimulated cycles in all patients having an embryo in the natural cycle, we found no difference in the distribution between the different cleavage stages. Of the cleaved embryos, 53% in the stimulated cycles had >or=4 cells versus 59% in the natural cycles after 2 days culture (P = 0.31). In the stimulated cycles 61% of the embryos had <10% fragmentation at the time of transfer on day 2, compared to 69% in the natural cycles (P = 0.15). CONCLUSION: The administration of exogenous gonadotrophins was not reflected in cleavage capacity or quality assessment of the resulting embryos.

Survival of primordial follicles following prolonged transportation of ovarian tissue prior to cryopreservation.

BACKGROUND: Cryopreservation of ovarian tissue for fertility preservation is becoming increasingly common. Treatment of diseases that may deprive the ovaries of follicles is often performed at local hospitals that are without the necessary facilities and expertise to cryopreserve ovarian tissue. The aim of the present study was to evaluate whether primordial follicles of ovarian cortex survive transport for up to 4 h prior to cryopreservation. METHODS: Immediately after recovery of one ovary from each of four patients, the cortex was roughly isolated, placed in IVF culture medium, kept on ice and transported for 3-4 h to the centre where final dissection and cryopreservation took place. Transplantation of pieces of thawed ovarian cortex under the skin of ovariectomized immunodeficient mice for a period of 4 weeks was used to assess the survival of primordial follicles. RESULTS: After transplantation, ovarian tissue from each of the four patients contained surviving follicles. CONCLUSIONS: Transport of roughly isolated ovarian cortex cooled on ice for a period of up to 4 h allows survival of primordial follicles following cryopreservation and transplantation to immunodeficient mice.

Th2-mediated atopic disease protection in Th1-mediated rheumatoid arthritis.

OBJECTIVE: The balance between CD4+ T-helper (h) cell subsets (Th1 and Th2) plays an important role in the pathogenesis of rheumatoid arthritis (RA) and atopy. While RA is believed to be a Th1 mediated disease, Th2 cells predominate in atopic disorders. The purpose of this study was to investigate differences in the occurrence of allergy, hay fever, house dust mite sensitivity and asthma, as well as total serum IgE levels in RA patients and controls. METHODS: The case history of atopic disorders was assessed in 134 RA patients and compared to those found in 305 healthy blood donors. RA patients also answered clinical questions concerning disease activity and severity. Total serum IgE levels were measured in both groups, taking into consideration disease modifying therapy. RESULTS: A significantly lower occurrence of medical history of hay fever (2.3%) and house dust mite sensitivity (3.1%) was found among RA patients compared to controls (24.2% and 12.2%, respectively; p < 0.0001 and p < 0.003 respectively). Moreover, RA patients had significantly lower total serum IgE levels than control subjects (p < 0.0001). RA was less severe in patients with atopy compared to non-atopic RA patients. CONCLUSION: These results support the concept that RA and atopy antagonize each other and that a change in the cytokine patterns of Th1 and Th2 cells could provide an indication for curative effects on RA.

Density and distribution of primordial follicles in single pieces of cortex from 21 patients and in individual pieces of cortex from three entire human ovaries.

BACKGROUND: At the time of cryopreservation of ovarian tissue for fertility preservation a small biopsy of ovarian cortex is usually taken for histological evaluation of the follicular reserve. The purpose of this study was to evaluate the distribution and density of primordial follicles in single pieces of cortex from individual patients and in pieces of cortex comprising entire ovaries, all prepared for cryopreservation. METHODS: Cortical biopsies from 21 patients and the whole cortex of one ovary were evaluated histologically prior to cryopreservation. In addition, the cortex of two whole ovaries was cryopreserved before histological evaluation. The volume of each cortical fragment was measured, all follicles counted and the follicular density calculated. RESULTS: In individual pieces of cortex follicular density showed a significant inverse linear correlation with age. The follicular density per cortical fragment prepared from each of the three entire ovaries varied from 1.8 to 166, 0.007 to 140 and 0.04 to 4.48 follicles/mm(3) cortical tissue. CONCLUSION: The density of primordial follicles varied more than two orders of magnitude in cortical fragments from each of the three ovaries. Primordial follicles were very unevenly distributed throughout the cortex of these ovaries, although a significant linear correlation between age and follicular density was found.

[Cervical myelopathy induced by calcium pyrophosphate tophus in primary chondrocalcinosis]. / Zervikale Myelopathie durch Kalzium-Pyrophosphat-Tophus bei primärer Chondrokalzinose.

A 78 year old female patient with cervical myelopathy induced by a calcium pyrophosphate dihydrate tophus around the dens axis with clinical signs of a cervical tumor is presented. After operation of the tumor, the diagnosis of a generalized CPPD disease was established retrospectively, where by patient had complained of joint pain already for many years. This known complication of a primary chondrocalcinosis should be a reason for a careful investigation of the cervical spine in CPPD disease, because neurological disturbances might increase the joint destruction in the manner of "Charcot joints".

Current aspects of colchicine therapy -- classical indications and new therapeutic uses.

Colchicine has been traditionally used for the treatment of gout. At present there is no generally accepted alternative to colchicine for the treatment of acute attacks or for the prevention of further attacks. The complex actions of this substance, which are mainly attributable to its stabilising action on the cytoskeleton and cell membranes, and its special pattern of distribution form the basis for the results presented here regarding the prophylactic or therapeutic actions of colchicine in a whole range of other diseases. This is all the more significant in that in several instances it concerns diseases that have so far been unsatisfactorily controlled by other treatments. Because of its astonishing absence of side effects, some authors consider that low dose colchicine may be considered as an alternative to previous therapies or even a means of first choice. It is therefore incorrect to think that medical research has shown little interest in this long-known potential and has not sought to confirm promising options by means of controlled studies. Fibrotic and inflammatory systemic diseases and those in which leucocytic chemotaxis play a role seem to be particularly predestined for this.

[Legal abortion. An analysis of factors which can affect frequency of complications]. / Legal abort. En analyse af faktorer, der kan have indflydelse på komplikationsfrekvensen.

INTRODUCTION: Due to a rather high complication rate, we decided to analyze data from women, who were readmitted to hospital with complications to legal abortion. MATERIAL: 760 legal abortions performed in 1993-1994 were retrospectively reviewed for complications. RESULTS: We found 66 complications (8.7% (95% confidence limits (6.7-10.7)), that resulted in readmission to hospital. The distribution of complications was as follows: retention 41 (62% (50-74)), infection 12 (18% (9-27)), bleeding and pain 11 (17% (8-26)) and perforation two (3% (0-7)). There was a slight correlation between a stronger midline echo on ultrasound measurement of the uterine cavity and finding placental tissue on histological examination of the material obtained from reevacuation. The time interval between legal abortion and reevacuation was nine days. Retroflexio uteri was not correlated with reevacuation. CONCLUSIONS: We found a statistically increased risk of complications following legal abortion relative to the figures reported by the national health authority, but not significantly higher than that found in other reports. Moreover, a conservative attitude to retention can reduce the number of reevacuatio uteri after legal abortion.

[Surgeons' experience of complication frequency--registration validity for legal abortion]. / Operatørerfaring i relation til komplikationsfrekvens--samt registreringsvaliditet ved legal abort.

INTRODUCTION: The aim was to investigate the degree to which experience influenced the complication rate (cr) in legal abortion. We also compared our cr with that registered by the National Health Service for the same group of patients. MATERIAL: 760 legal abortions from 1993 to 1994 were reviewed retrospectively. RESULTS: We found an overall cr of 8.7% (95% confidence limit: 6.7-10.7). The cr was the same for all groups of doctors, from the youngest to the most senior and experienced surgeons. The experience in months of the younger surgeons was noted for each intervention, and we found that they did not improve their results over time or by practice. In the same study, we found that the National Health Service had received, registered, and published statistical data from the hospitals, which were not entirely accurate. CONCLUSION: The complication rate was not associated with the experience of the surgeon.