Characterization of the thymus in Lrp4 myasthenia gravis: Four cases

Myasthenia gravis (MG) is an autoimmune disease of the neuromuscular junction. Most patients have pathogenic autoantibodies against the acetylcholine receptor (AChR). In the last years a novel subpopulation of MG patients has been described that harbors antibodies against low-density lipoprotein receptor-related protein 4 (Lrp4), another postsynaptic neuromuscular antigen. In early-onset AChR MG (EOMG), the thymus plays an important role in immunopathogenesis, and early thymectomy is beneficial. It is still unknown if the thymus plays any role in Lrp4-MG. In this pilot study, we compared thymus samples from four patients with Lrp4-MG (one pre-treated with immunosuppressive drugs), four non-MG controls and five EOMG patients (not pretreated with immunosuppressive drugs). Immunohistochemistry of the Lrp4-MG thymi revealed normal architecture, with normal numbers and distribution of B-cells, lymphoid follicles and Hassall's corpuscles. Primary CD23⁺ lymphoid follicles were similarly infrequent in Lrp4-MG and control thymic sections. In none of the control or Lrp4-MG thymi did we find secondary follicles with CD10⁺ germinal centers. These were evident in 2 of the 5 EOMG thymi, where primary lymphoid follicles were also more frequent on average, thus showing considerable heterogeneity between patients. Even if characteristic pathological thymic changes were not observed in the Lrp4 subgroup, we cannot exclude a role for the thymus in Lrp4-MG pathogenesis, since one Lrp4-MG patient went into clinical remission after thymectomy alone (at one year follow-up) and one more improved after thymectomy in combination with immunosuppressive therapy.     

New molecular diagnostic trends and biomarkers for amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis (ALS) is a rare and fatal neurodegenerative disorder. Two forms are recognized, familial (FALS) that accounts for 5–10% of ALS cases, and sporadic (SALS) that accounts for the rest. Early diagnosis of ALS is important because it improves their therapeutic efficacy. Current diagnosis is based on clinical assessment and requires approximately 12 months, leading to a significant delay in drug administration. Therefore, new methods are required for the earlier diagnosis of ALS. Screening for pathogenic variants in known ALS‐associated genes is already exploited as a diagnostic tool in ALS but cannot be applied for population‐based screening. New circulating biomarkers (proteins or small molecules) are needed for initial screening, whereas specific diagnostic methods can be applied to confirm the presence of pathogenic variants in the selected population subgroup. Lipids appear as promising biomarkers for population‐based screening and for monitoring disease progression. Genetic analysis can also assist in the prediction of disease progression by analyzing disease‐modifying genes, for example, EPHA4 and CHGB. Furthermore, molecular diagnosis will aid the stratification of ALS patients for improved pharmacological approaches. Here, we discuss current and novel diagnostic strategies and how they can be applied to revolutionize the field of ALS molecular diagnosis.     

Periodontitis associated with Hajdu-Cheney syndrome

BACKGROUND: Hajdu-Cheney syndrome (HCS) is an inheritable, rare disorder of bone metabolism, associated with acro-osteolysis of the distal phalanges, short stature, distinctive craniofacial and skull changes, premature tooth loss, and periodontitis. This report focuses on the periodontal manifestations of HCS. METHODS: A 22-year-old female presented with the characteristic clinical features of HCS, including short stature, small face, prominent epicanthal folds, thin lips, small mouth, and short hands. There were no abnormal biochemical, hematological, or hormonal data. Tests for bone mineral density were indicative of osteoporosis. Cephalometric analysis revealed hypoplasia of the midface and increased cranial base angle; the maxilla and the mandible were set posteriorly. The sella turcica was enlarged, elongated, and wide open with slender clinoids. Hair samples were examined by scanning electron microscopy, and tooth cementum and dentin were evaluated histologically. RESULTS: According to the periodontal evaluation, gingival inflammation was 12.5%, bleeding on probing score was 24%, probing depths averaged 4 to 6 mm, and clinical attachment loss averaged 3 to 6 mm. Class II furcations were found on three teeth. Almost all teeth exhibited pathological mobility of varying degrees. There was a generalized, horizontal bone loss of approximately 50%. Three teeth had to be extracted because of severe localized periodontal destruction. Histologic examination of the dentin and the cementum was normal. CONCLUSIONS: HCS periodontitis is associated with an unpredictable and uneven, rapid rate of periodontal destruction of unknown etiology. Further research is required to identify the role of the possible pathogenic factors involved.     

Angiogeneseinhibition xenotransplantierter Plattenepithelkarzinome

Clinical and histopathological results in human tumors indicate that the connection of solid tumors to the vascular system precedes the exponential tumor growth and further progression. Acknowledging the concept of tumor angiogenesis, the search for angiogenesis inhibiting agents as potential drugs in cancer treatment began rather early. In the present preclinical nude mice model, the antitumoral effect of TNP-470 on xenotransplanted squamous cell carcinoma was tested. The chosen dosage of 30 mg/kg and 60 mg/kg resulted in a significant growth inhibition (P = 0.006 and P = 0.01) compared to the control group. The available in vivo and in vitro data lead to the conclusion that the concept of angiogenesis inhibition will have some impact on treatment of solid tumors in the near future.     

Solid cerebral echinococcosis mimicking a primary brain tumor

Solitary brain affection is rare in echinococcosis. We report the case of a 35-year-old woman presenting with symptomatic grand-mal epilepsy due to a right frontal, partially cystic space-occupying lesion. Pre-operative computed tomography and magnetic resonance imaging (MRI) suggested a cystic astrocytoma. However, histological examination yielded the diagnosis of a 'chitinoma', a rare subtype of solid cerebral hydatid disease (echinococcosis). It mimicked a primary brain tumor and, therefore, posed a diagnostic problem. We present the--to our knowledge--first MRI scans in a case of a histologically proven chitinoma.     

Influence of advanced age on the blastocyst development rate and pregnancy rate in assisted reproductive technology

Objective: To evaluate the percentage of blastocysts developing, the pregnancy rate, the implantation rate, and the abortion rate in women >40 years of age using a cell-free culture system for the development of viable human blastocysts.

Design: Retrospective clinical study.

Setting: Private IVF units.

Patient(s): Two hundred ninety-three cycles in patients undergoing IVF treatment for infertility. Sixty-two cycles were in patients ≥40 years of age, and 231 cycles were in patients <40 years of age.

Intervention(s): Pronucleate oocytes obtained from IVF were cultured in vitro for 5–6 days. One to four embryos were transferred.

Main Outcome Measure(s): Blastocyst development rate, pregnancy rate, implantation rate, and abortion rate.

Result(s): From 293 cycles, 3,115 pronucleate oocytes were cultured, producing 1,175 blastocysts. In the women >40 years of age, the blastocyst development rate was 22.2%, and in the younger group, the rate was 40.5%. The pregnancy rate and implantation rate in the ≥40-year age group were 21.1% and 8.9%, respectively; corresponding rates in the younger group were 44.6% and 19.9%. The abortion rate was increased for the ≥40-year age group (25% versus 13.3%).

Conclusion(s): Success rates for the development of viable human blastocysts, pregnancy, and implantation decline significantly in women ≥40 years old.     

p53 and bcl-2 protein expression in rectosigmoid adenomas

Objective: The aim of this study was to evaluate whether p53 or bcl-2 protein expression in rectosigmoid adenomas is associated with histological characteristics of the adenomas and with presence of synchronous advanced proximal neoplasms.
Methods: Seventy-six average-risk patients who underwent total colonoscopy and had rectosigmoid adenoma(s) were studied. An adenoma was considered advanced if villous histology and/or severe dysplasia and/or diameter >1 cm were present. p53 And bcl-2 protein expression was immunohistochemically examined using specific monoclonal antibodies.
Results: p53 Protein was detected in 43% and bcl-2 in 93% of the 76 rectosigmoid adenomas. Advanced compared with nonadvanced adenomas were significantly more frequently p53-positive (28 of 44 or 63.6% vs five of 32 or 15.6%,   p < 10⁻⁴  ) or had a bcl-2 score of 12 (20 of 44 or 45.5% vs five of 32 or 15.6%,   p = 0.007  ). Proximal advanced neoplasms were mainly found in patients with rectosigmoid adenomas positive for p53 and with a bcl-2 score of 12 (six of 17 or 35.3% vs 2/59 or 3.4%, OR: 15.6,   p = 0.001  ) and in particular in those with advanced rectosigmoid adenomas positive for p53 and with a bcl-2 score of 12 (six of 13 or 46.2% vs two of 31 or 6.5%, OR: 12.4,   p = 0.007  ).
Conclusion: p53 Expression and bcl-2 protein overexpression in rectosigmoid adenomas are associated with advanced histology and a high risk of synchronous advanced proximal colon neoplasm.     

What Has Changed in the Management of Uterine Serous Carcinomas? Two Decades of Experience

Uterine serous carcinoma accounts for 3–10% of endometrial cancers, but it is the most lethal histopathological subtype. The molecular characterization of endometrial carcinomas has allowed novel therapeutic approaches for these patients. We undertook a retrospective analysis of patients with uterine serous carcinomas treated in our hospital within the last two decades to identify possible changes in their management. The patients and their characteristics were evenly distributed across the two decades. Treatment modalities did not change significantly throughout this period. After adjuvant treatment, patients’ median disease-free survival was 42.07 months (95% CI: 20.28–63.85), and it did not differ significantly between the two decades (p = 0.059). The median overall survival was 47.51 months (95% Cl: 32.18–62.83), and it significantly favored the first decade’s patients (p = 0.024). In patients with de novo metastatic or recurrent disease, median progression-free survival was 7.8 months (95% Cl: 5.81–9.93), whereas both the median progression-free survival and the median overall survival of these patients did not show any significant improvement during the examined time period. Overall, the results of our study explore the minor changes in respect of uterine serous carcinoma’s treatment over the last two decades, which are reflected in the survival outcomes of these patients and consequently underline the critical need for therapeutic advances in the near future.     

Optimisation of bacterial release from a stable microfluidic-generated water-in-oil-in-water emulsion

Application of droplet microfluidics for the encapsulation of bacteria in water-in-oil-in-water (W/O/W) emulsion allows for production of monodisperse droplets with controllable size. In this study the release of bacteria from W/O/W emulsion, the effect of the double emulsion structure on bacterial growth and metabolic activity, and the stability and mechanism of bacterial release were investigated. W/O/W emulsions were formed using a double flow-focusing junction microfluidic device under controlled pressure to produce droplets of approximately 100 μm in diameter containing an inner aqueous phase (W₁) of about 40–50 μm in diameter. GFP-labelled Escherichia coli (E. coli-GFP) bacteria were encapsulated within the W₁ droplets and the stability of emulsions was studied by monitoring droplet size and creaming behaviour. The double emulsions were stabilised using a hydrophilic (Tween 80) and a lipophilic surfactant (polyglycerol polyricinoleate) and were destabilised by altering the osmotic balance, adding NaCl either in the inner W₁ phase (hypo-osmotic) or outer W₂ phase (hyper-osmotic). The release of E. coli-GFP was monitored by plating on agar whereby the colony form unit (CFU) of the released bacteria was determined while fluorescent microscopy was employed to observe the mechanism of release from the droplets. The release of E. coli-GFP was significantly increased with higher concentrations of NaCl and lower amounts of Tween 80. Microscopic observation revealed a two-step mechanism for the release of bacteria: double W/O/W emulsion droplet splitting to release W₁ droplets forming a secondary double emulsion followed by the collapse of W₁ droplets to release E. coli-GFP into the continuous aqueous phase.

Encapsulation enhanced viability and metabolic activity. Nutrients can cross the oil layer. Bacterial release increased while emulsion stability decreased at high osmotic pressure and low surfactant concentration. Two-step release mechanism observed.