ELISAs for free light chains of human immunoglobulins using monoclonal antibodies: comparison of their specificity with available polyclonal antibodies

For the measurement of human immunoglobulin free light chains (LCs) in clinical samples, highly specific assays for free LCs are required to discriminate them from LC portions of intact immunoglobulins (bound LCs). To develop specific enzyme-linked immunosorbent assays (ELISAs) for free LCs, two anti-free LC kappa and lambda monoclonal antibodies (MAbs) were raised by a mouse/mouse hybridoma technique. We compared the specificities of these two MAbs with those of six commercially available anti-free LC antisera, which are widely used in free LC immunoassays. Comparative titrations against free LCs and intact IgGs showed the MAbs had less cross-reactivity to intact IgGs, while the four of six antisera had high reactivity to intact IgGs. Using these MAbs, we developed LC kappa and LC lambda ELISAs with ranges from 7.8 to 500 micro g/l of free LCs and less cross-reactivity to intact IgGs (less than 0.12%). On the other hand, ELISAs with anti-free LC antisera showed low specificity and/or sensitivity. Thus, the use of these MAbs may provide reliable methods for specific measurements of free LCs in clinical samples.     

Incidences and risk factors of moderate-to-severe ovarian hyperstimulation syndrome and severe hemoperitoneum in 1,435,108 oocyte retrievals

Research questionWhat are the risk factors affecting the incidences of moderate-to-severe ovarian hyperstimulation syndrome (OHSS) and severe hemoperitoneum in assisted reproductive technology (ART) treatment cycles?DesignA retrospective cohort study was conducted on 1,435,108 oocyte retrieval cycles among Japanese ART registry data between 2007 and 2015. The study included 11,378 cycles with moderate-to-severe OHSS, 1182 cycles with severe hemoperitoneum, including 27 cycles with both conditions, and 1,422,575 cycles without moderate-to-severe OHSS and severe hemoperitoneum.ResultsThe incidences of moderate-to-severe OHSS and severe hemoperitoneum were 0.79% and 0.08%, respectively, and decreased by 0.57-fold and 0.29-fold from 2007 to 2015, respectively. In cycles with OHSS and cycles with hemoperitoneum women were younger (odds ratios [OR] 0.91 and 0.95, respectively) and had more retrieved oocytes (OR 1.09 and 1.01, respectively) compared with cycles without both complications. The use of a gonadotrophin-releasing hormone (GnRH) agonist protocol for ovarian stimulation was the highest risk factor in cycles with OHSS and hemoperitoneum (OR 1.83 and 1.24, respectively), followed by GnRH antagonist protocol (reference), gonadotrophin with or without oral medicine (OR 0.45 and 0.56, respectively) and natural or oral medicine (OR 0.02 and 0.19, respectively). In fresh embryo transfer, clinical pregnancy was associated with an increased risk of OHSS and hemoperitoneum (OR 1.19 and 2.34, respectively).ConclusionsThe highest risk factors affecting OHSS and hemoperitoneum were the use of a GnRH agonist protocol and clinical pregnancy following fresh embryo transfer. The incidences of OHSS and hemoperitoneum have decreased yearly with a reduction of GnRH agonist use and fresh embryo transfer.     

An electron microscopic study of glomerular lesions in hereditary nephrotic mice (ICGN strain)

Summary Glomerular lesions in hereditary nephrotic mice (ICGN strain) were investigated by electron microscopy. The glomeruli of unaffected animals, which appeared normal by light microscopy, had developed an ultrastructural change in the glomerular capillary basement membrane (GCBM). There was a partial thickening of the GCBM with bilaminar splitting of the lamina densa and an electron-dense fibrillar material exhibiting cross-striations. In affected animals, light microscopy revealed a marked thickening of GCBM and an increase of mesangial matrix without cellular proliferaton. By electron microscopy, multilaminar splitting of the lamina densa in the thickened GCBMs and fusion of the epithelial foot processes were observed. In some severely affected animals, immune complex deposition was found in GCBM, but little if any was observed in other animals. In the end, the glomeruli were globally sclerosed. Our findings suggest that initial structural abnormalities in GCBM may play an important role in the onset and development of the disease, though subsequent events such as immune complex deposition would modify the disease.     

Study of Vimentin Expression in Non-Hodgkin's Lymphoma Using Paraffin Sections

Human non-Hodgkin's lymphomas were studied by means of an avidin biotin complex immunoperoxidase method using several monoclonal antibodies against the intermediate filament protein, vimentin. The study cases were 61 B cell lymphomas (including 2 plasmacytomas) and 30 T cell lymphomas (including 8 cases of mycosis fungoides). Twelve of the 61 B cell lymphomas were positive for vimentin, and were composed of extrafollicular center cells such as immunoblastic and plasmacytoid cells. On the other hand, lymphomas of follicular center cell origin were negative for vimentin. All cases of T cell lymphoma except for 14 (all of 9 AlLD- type lymphomas, all of 4 lymphoblastic lymphomas and one diffuse mixed small/ large lymphoma) were positive for vimentin. Although vimentin expression appeared to be influenced by various conditions such as the proportion of T- and B cell subsets, or B cell proliferation rate, follicular center cells were constantly negative for vimentin.     

Anterior mediastinal tumors: diagnostic accuracy of CT and MRI

Purpose

To compare the diagnostic accuracy for anterior mediastinal tumors among CT, MRI, and both CT and MRI, and to determine the optimal CT and MRI procedures for the diagnosis of anterior mediastinal tumors.

Materials and methods

Both CT and MRI were performed in 127 patients with pathologically diagnosed anterior mediastinal tumors. The patients included 48 cases of thymoma, 12 cases of thymic carcinoma, 12 cases of thymic cyst, 20 cases of mature teratoma, 13 cases of malignant germ cell tumor, and 22 cases of malignant lymphoma. The CT and MRI scans were assessed by two chest radiologists without knowledge of their clinical and pathologic data. The observers recorded various CT and MRI findings and their first choice of diagnosis.

Results

The two observers made a correct first-choice diagnosis in an average of 78 (61%) of 127 cases on CT, 71 (56%) of 127 cases on MRI, and 85.5 (67%) of 127 cases on both CT and MRI. These included 83% cases of thymoma on CT, 84% on MRI, and 85% on both CT and MRI; 38% cases of thymic carcinoma on CT and 13% on MRI, and 33% on both CT and MRI; 46% cases of thymic cyst on CT and 71% on MRI, and 63% on both CT and MRI; 58% cases of mature teratoma and 38% on MRI, and 78% on both CT and MRI; 35% cases of malignant germ cell tumor on CT and 27% on MRI, and 31% on both CT and MRI; and 55% cases of malignant lymphoma on CT and 43% on MRI, and 61% on both CT and MRI. There were significant differences between the diagnostic accuracy by CT and MRI in the cases with both thymic cysts and thymic carcinoma (p < 0.05).

Conclusion

CT is equal or superior to MRI in the diagnosis of anterior mediastinal tumors except for thymic cyst. CT should be considered the modality of choice following chest radiography, however, in certain circumstances, such as thymic cyst with hemorrhage or inflammation which mimic solid tumor despite low enhancement, MRI may be better in distinguishing anterior mediastinal tumors. For more helpful information in the diagnosis of mature teratoma after CT, MRI may follow.     

Clinical and neuroendocrinological characteristics of delayed orthostatic hypotension in Parkinson’s disease

Clinical Autonomic Research - Delayed orthostatic hypotension (DOH), a fall in blood pressure after a 3-min cutoff, is clinically meaningful. The aim of this study was to elucidate the clinical and...     

A novel stop-gain CUL3 mutation in a Japanese patient with autism spectrum disorder

BackgroundCUL3 encodes cullin-3, a core component of a ubiquitin E3 ligase. CUL3 mutations have recently been associated with autism spectrum disorder (ASD); however, the detailed clinical courses have been described in only a limited number of patients with CUL3 mutations and neurodevelopmental diseases, including ASD.Case reportA 21-month-old Japanese girl presented with febrile status epilepticus and thereafter exhibited developmental regression, including loss of her verbal ability, eye contact, and skills in activities of daily living. Trio-based exome sequencing identified a de novo two-base insertion in CUL3, c.1758_1759insTG, p.(Thr587*).ConclusionWe report a case of a patient with ASD and a stop-gain CUL3 variant. Screening of CUL3 variants is worth considering for patients with ASD, especially those with Rett-like developmental regression.     

The temporal profile of the reaction of microglia,astrocytes, and macrophages in the delayed onset paraplegia after transient spinal cord ischemia in rabbits

In the present study, we sought to elucidate the temporal profile of the reaction of microglia, astrocytes, and macrophages in the progression of delayed onset motor dysfunction after spinal cord ischemia (15 min) in rabbits. At 2, 4, 8, 12, 24, and 48 h after reperfusion (9 animals in each), hind limb motor function was assessed, and the lumbar spinal cord was histologically examined. Delayed motor dysfunction was observed in most animals at 48 h after ischemia, which could be predicted by a poor recovery of segmental spinal cord evoked potentials at 15 min of reperfusion. In the gray matter of the lumbar spinal cord, both microglia and astrocytes were activated early (2 h) after reperfusion. Microglia were diffusely activated and engulfed motor neurons irrespective of the recovery of segmental spinal cord evoked potentials. In contrast, early astrocytic activation was confined to the area where neurons started to show degeneration. Macrophages were first detected at 8 h after reperfusion and mainly surrounded the infarction area later. Although the precise roles of the activation of microglia, astrocytes, and macrophages are to be further determined, the results indicate that understanding functional changes of astrocytes may be important in the mechanism of delayed onset motor dysfunction including paraplegia. IMPLICATIONS: Microglia and macrophages play a role in removing tissue debris after transient spinal cord ischemia. Disturbance of astrocytic defense mechanism, breakdown of the blood-spinal cord barrier, or both seemed to be involved in the development of delayed motor dysfunction.     

Preliminary results of intermittent retrograde cerebral perfusion during proximal aortic arch surgery.

OBJECTIVE: Continuous retrograde cerebral perfusion during aortic arch surgery is associated with cerebral edema. In this report, we describe the clinical use of a new type of intermittent retrograde cerebral perfusion. SUBJECTS AND METHODS: Fourteen patients with a Stanford type A dissection were included in this study. With the usual method of retrograde cerebral perfusion, about 2,500 mL venous blood is drained from bicaval cannulae into a hard-shell reservoir, and oxygenated blood is perfused through the superior vena caval cannula. The flow rate is 300 mL/min. After about 15 min, retrograde perfusion is discontinued, and drainage from the bicaval cannulae is restarted. When a bloodless field is necessary, perfusion also is discontinued. RESULTS: Two to seven cycles of intermittent retrograde cerebral perfusion were administered (average, 3.1+/-0.4, mean+/-SD). The total retrograde perfusion time was 36.0+/-1.9 min which was equivalent to 74.8% of the circulatory arrest time. No patient developed edema of the upper body. The time to wake-up was 3 to 14 h (average, 6.5+/-1.0 h). No patient suffered any neurologic complications even though the time of circulatory arrest was greater than 60 min in four cases. Head magnetic resonance imaging or computed tomography was performed in 12 cases, and no evidence of hypoxic brain injury was detected. CONCLUSIONS: Our clinical experience using a moderate amount of intermittent retrograde cerebral perfusion is superior to continuous retrograde cerebral perfusion for protecting the brain during aortic arch surgery.     

Folic acid reduces risks of having fetus affected with neural tube defects: dietary food folate and plasma folate concentration

OBJECTIVES: Risk of having fetus affected with neural tube defects can be reduced by maternal periconceptional folic acid supplementation. The purpose of the present study is to investigate how folate is taken from diets and to measure plasma folate concentrations. SUBJECTS AND METHODS: A total of 222 women comprising 5 groups, i.e., healthy women, mothers of myelodysplastic patients, pregnant women, myelodysplastic patients, nurse students, participated in our study. Food frequency questionnaires kept 3 days were analyzed based on the 5th standard table of food composition in Japan. Plasma folate concentrations were measured by means of chemiluminescent immunoassay method. Changes in plasma folate concentrations and possible adverse effects following the folic acid supplementation for 16 weeks were also investigated. RESULTS: The dietary intake of folate, plasma folate concentration and energy intake averaged 293 micrograms/day, 8.1 ng/ml and 1,857 Kcal, respectively, among the subjects. Pregnant women took the largest amount of folate from diets and demonstrated the highest plasma folate concentration among the groups. The dietary folate in myelodysplastic patients and nurse students was significantly lower compared to that of healthy women. The Recommended Dietary Allowance of folate was not fulfilled in 22% of non-pregnant adult women and 72% of pregnant women. The dietary folate was mainly taken from the 3rd food group but the 4th group of food was consumed most. Mean folate intake was significantly correlated with circulating concentrations of serum folate (p = 0.012 r = 0.186). The consecutive administration of 400 micrograms supplements for 16 weeks increased a baseline plasma value of 8.7 ng/ml to 32.6 but fell down rapidly to 17.3 24 hours later without any adverse effects. CONCLUSIONS: The dietary folate and serum folate concentrations averaged 293 micrograms/day and 8.1 ng/ml, respectively. The former is the first report based on the 5th standard table of food composition in Japan. Majority of pregnant women took less dietary folate than what recommended by the government. Those who are capable of becoming pregnant are recommended to consume much of the 3rd food group and those who are planning to become pregnant are recommended to take 400 micrograms of folic acid supplements from 4 weeks before to 12 weeks after conception.