Two parts of the nucleus prepositus hypoglossi project to two different subdivisions of the dorsolateral periaqueductal gray in cat

The dorsolateral column of the mesencephalic periaqueductal gray (PAG) is a separate part of the PAG. Its afferent sources, efferent targets, and neurochemical properties differ from the adjacent PAG columns. The dorsolateral PAG is thought to be associated with aversive behaviors, but it is not yet understood how these behaviors are brought about. To elucidate the function of the PAG further, in the present study we investigated which brainstem regions project to the dorsolateral PAG. Wheat germ agglutinin-horseradish peroxidase (WGA-HRP) injections involving the dorsolateral PAG, but extending into the lateral part, resulted in many retrogradely labeled cells in the pontine and medullary tegmentum bilaterally. However, it was concluded that these neurons were labeled from the lateral PAG, because no anterograde labeling was found in the dorsolateral PAG after a large injection into the tegmentum. Retrogradely labeled cells were also found in the nucleus prepositus hypoglossi (PPH), mainly contralaterally. Injections of [3H]leucine or WGA-HRP in the PPH resulted in anterogradely labeled fibers in the dorsolateral PAG. Two separate distribution patterns were found. The caudal and intermediate PPH projected to a small region on the dorsolateral edge of the dorsolateral column, whereas the supragenual PPH distributed labeled fibers to all other parts of the dorsolateral PAG, except the area on the dorsolateral edge. These separate PPH projections suggest that two subdivisions exist within the dorsolateral PAG. The present findings suggest a role for the dorsolateral PAG in the oculomotor system.     

Segmental and laminar organization of the spinothalamic neurons in cat: evidence for at least five separate clusters

The spinothalamic tract (STT), well known for its role in the relay of information about noxe, temperature, and crude touch, is usually associated with projections from lamina I, but spinothalamic neurons in other laminae have also been reported. In cat, no complete overview exists of the precise location and number of spinal cells that project to the thalamus. In the present study the laminar distribution of retrogradely labeled cells in all spinal segments (C1-Coc2) was investigated after large WGA-HRP injections in the thalamus. The results show that this distribution of STT cells differed greatly between the different spinal segments. Quantitative analysis showed that there exist at least five separate clusters of spinothalamic neurons. Lamina I neurons in cluster A and lamina V neurons in cluster B are mainly found contralaterally throughout the length of the spinal cord. Cluster C neurons are located bilaterally in the ventrolateral part of laminae VI-VII and lamina VIII of the C1-C3 spinal cord. Cluster D neurons were found contralaterally in lamina VI in the C1-C2 segments, and cluster E neurons were located mainly contralaterally in the medial part of laminae VI-VII and lamina VIII of the lumbosacral cord. Most spinothalamic neurons are not located in the enlargements and most spinothalamic neurons are not located in lamina I, as suggested by several other authors. The location of the spinothalamic neurons shows remarkable similarities, but also differences, with the location of spino-periaqueductal gray neurons.     

Mental illness and health care use: a study among new neurological patients

We investigated 198 new neurological patients to learn if mental illness had an impact on nonpsychiatric health care use. Mental illness was assessed in a two-phase design, including the Schedules for Clinical Assessment in Neuropsychiatry (SCAN), providing ICD-10 diagnoses and two brief rating scales: the Symptom Check List (SCL-8) for anxiety and depression and the Whiteley-7 for somatization. The patients' use of health care was studied 5-10 years before and 18 months after the psychiatric assessment, using data from national patient registers. Mental illness was linked to data on health care use through weighted logistic regression. ICD-10 psychiatric disorders increased the risk of subsequent high use of nonpsychiatric hospital admissions - somatoform disorders with more than five times (OR=5.6; 95% CI=1.6-20.1) and anxiety/depression with almost four times (OR=3.7; 95% CI=1.1-12.0). There was also a trend, though less marked, linking mental illness to previous hospital use. Use of primary care was also markedly increased by mental disorders, however, only in patients entering the study as inpatients. In conclusion, neurological patients are at risk of being high users of health care if they have a mental illness, somatoform disorders being the most powerful risk factor. The results are consistent with the findings among internal medical patients.     

Prenatal stress may increase vulnerability to life events: comparison with the effects of prenatal dexamethasone

Prenatal stress has been associated with a variety of alterations in the offspring. The presented observations suggest that rather than causing changes in the offspring per se, prenatal stress may increase the organism's vulnerability to aversive life events. Offspring of rat dams stressed gestationally by chronic mild stress (CMS, a variable schedule of different stressors) or dexamethasone (DEX, a synthetic glucocorticoid, i.e., a pharmacological stressor) was tested for reactivity by testing their acoustic startle response (ASR). Two subsets of offspring were tested. One was experimentally na?ve at the time of ASR testing, whereas the other had been through blood sampling for assessment of the hormonal stress response to restraint, 3 months previously. Both prenatal CMS and dexamethasone increased ASR in the offspring compared to controls, but only in prenatally stressed offspring that had been blood sampled 3 months previously. In conclusion, similarity of the effects of maternal gestational exposure to a regular stress schedule and of exposure to a synthetic glucocorticoid suggests that maternal glucocorticoids may be a determining factor for changes in the regulatory mechanisms of the acoustic startle response. Further, a single aversive life event showed capable of changing the reactivity of prenatally stressed offspring, whereas offspring of dams going through a less stressful gestation was largely unaffected by this event. This suggests that circumstances dating back to the very beginning of life affect the individual's sensitivity towards experiences in life after birth. The prenatal environment may thus form part of the explanation of the considerable individual variation in the development of psychopathology.     

Clinical-scale radiolabeling of a humanized anticarcinoembryonic antigen monoclonal antibody, hMN-14, with residualizing 131I for use in radioimmunotherapy.

Radiolabeling of monoclonal antibodies (mAbs) with an intracellularly trapped form of (131)I (residualizing (131)I) involves radioiodinating a small molecular entity, conjugating it to the mAb, and purification. Column purifications are impractical during procedures involving multi-gigabecquerel levels of radioactivity. The goal of this study was to develop a simple, remote, "1-pot" method of radiolabeling and purification for the scaled-up radioiodination of a humanized anti-carcinoembryonic antigen (CEA) mAb, humanized MN-14 (hMN-14; labetuzumab), with an optimized residualizing (131)I moiety, (131)I-IMP-R4. IMP-R4 is MCC-Lys(MCC)-Lys(X)-d-Tyr-d-Lys(X)-OH, where MCC is 4-(N-maleimidomethyl)-cyclohexane-1-carbonyl and X is 1-((4-thiocarbonylamino)benzyl)-diethylenetriaminepentaacetic acid. METHODS: An IODO-GEN-based remote labeling system was used. IMP-R4 was radioiodinated (0.13 mumol per 3.7 GBq of (131)I) at a pH of 7.0-7.4 and conjugated to disulfide-reduced hMN-14 after quenching of unused reactive (131)I. The product was purified by stirring for 5 min with a 20% (w/v) suspension of an anion-exchange resin and sterilely filtered into a sealed vial. Human serum albumin was added at a final concentration of 1%-2.5%. Immunoreactivity was determined by mixing with CEA and determining the complexation level by size-exclusion high-pressure liquid chromatography. Two control radiolabelings, either with unreduced hMN-14 or with IMP-R4 omitted, also were performed. RESULTS: In 18 radiolabelings with (131)I in the range of 2.04-4.81 GBq (55-130 mCi), yields of 59.9% +/- 7.9% (mean +/- SD) at specific activities of 200 +/- 26 MBq/mg (5.4 +/- 0.7 mCi/mg) were obtained, with > or =95% of the radioactivity being associated with hMN-14 and with < or =4% aggregation. Similar yields were obtained in a subset of radiolabelings (n = 7) with >3.7 GBq of (131)I. The immunoreactivities of the preparations were typically >95%. Nonspecific incorporation in the absence of IMP-R4 was 0.5%, whereas that obtained with unreduced IgG was approximately 8%, possibly because of conjugation of IMP-R4 at lysine sites. The process also removed >99% of the quenching reagent used. Radiolabelings performed with freshly prepared solutions or lyophilized preparations produced similar yields, a result that suggested the option for a single-use kit design. CONCLUSION: Efficient removal of (131)I-IMP-R4 and quenched (131)I by 5 min of stirring with anion-exchange resin renders a multi-gigabecquerel-level preparation of (131)I-IMP-R4-hMN-14 safe, convenient, and practical.     

High levels of neutralizing IL-6 autoantibodies in 0.1% of apparently healthy blood donors

IL-6-specific autoantibodies (aAb-IL-6) have been reported in diseased and healthy individuals. We recently established a model for aAb-IL-6 in different mouse strains, based on vaccination with immunogenic IL-6 analogues, in which titers of aAb-IL-6 above 1,000 resulted in an in vivo IL-6 deficiency. Here, we examined aAb-IL-6 in 4,230 blood donors. Stable low titers of aAb-IL-6 were found in 9% of the donors, while 1% had titers ranging from 64 to greater than 10,000. Such aAb-IL-6-positive donors appeared normal with no overt signs of pathology. Natural and recombinant forms of IL-6 bound avidly to their IgG, and their plasma strongly neutralized IL-6 in vitro. Slightly elevated concentrations of IL-6 exclusively in the form of IL-6-IgG complexes were present in their circulation. The complexes did not contain soluble IL-6 receptors. Titers of 0.1% of the blood donors were as positive as the vaccination-induced IL-6-deficient mice. Such donors might be IL-6 deficient, and if so, IL-6 seems be dispensable for several months in otherwise healthy individuals. Such highly positive donors also explain why normal human IgG for pharmaceutical use may contain high anti-IL-6 activity. Finally, transfusion of plasma with a high titer of aAb-IL-6 might, temporally, render a recipient IL-6 deficient.     

Selective killing of B-cell hybridomas targeting proteinase 3, Wegener's autoantigen

Wegener's granulomatosis (WG) is a rare disease characterized by granulomatous lesions, small vessel vasculitis and the presence of anti-neutrophil cytoplasmic autoantibodies (C-ANCAs) in the sera of affected patients. Their main target antigen is proteinase 3 (PR3), a neutrophil and monocyte-derived neutral serine protease. Since the standard treatment of this severe autoimmune disease, with cyclophosphamide and corticosteroids, is associated with potential side-effects, the development of a more specific immunotherapeutic agent is warranted. The key role of ANCA in the pathogenesis of vasculitis and the effectiveness of anti-CD20 antibodies in patients with refractory WG points towards the importance of B cells in WG. We thus evaluated a new approach to selectively eliminate PR3-specific autoreactive B cells by targeting the B-cell receptor. For this purpose we used a bifunctional recombinant fusion protein consisting of the antigen PR3 and a toxin. The cytotoxic component of this novel fusion protein was the ribonuclease angiogenin, a human toxin with low immunogenicity. The toxin was stabilized by exchanging the catalytically relevant histidine in position 44 with glutamine to eliminate the autoproteolytic activity. PR3H44Q was fused either to the N terminus or to the C terminus of angiogenin. The recombinant proteins were expressed in 293T cells. Binding assays demonstrated the appropriate size and recognition by anti-PR3 antibodies. Using TUNEL technology, we demonstrated that these autoantigen toxins kill proteinase 3-specific B-cell hybridomas selectively by inducing apoptosis. The data indicate that autoantigen-toxins are promising tools in the treatment or co-treatment of autoimmune diseases in which the antigen is known.     

The natural history of primary dysmenorrhoea: a longitudinal study

Objective   To describe the prevalence, severity, course and predictive factors of primary dysmenorrhoea in women of all reproductive ages.
Design   Prospective mailed surveys in 1985 and 1991.
Setting   University of Iowa, College of Nursing.
Population   We began with a stratified sample of 996 nurses who graduated between 1963 and 1984. We analysed data from 404 women who responded to both surveys, but denied endometriosis, pelvic inflammatory disease or uterine fibroids.
Methods   Participants were surveyed twice at an interval of six years (response rates 73% and 78%) regarding menstrual cycle characteristics. For analysis, dysmenorrhoea was dichotomised as none/mild or moderate/severe. We analysed predictive factors using χ² tests and stepwise logistic regression.
Main outcome measure   Severity of dysmenorrhoea. Menstrual cramps as experienced when not taking medication to prevent discomfort were rated on a four-point scale: 0 = no dysmenorrhoea, 1 = minimal (can work, somewhat uncomfortable), 2 = moderate (can work, but quite uncomfortable) or 3 = severe dysmenorrhoea (miss work, have to be in bed).
Results   In 1985, 80% of respondents were >25 years old and 60% were parous. There were few changes over six years in the prevalence of mild (51% to 53%), moderate (22% to 20%) or severe dysmenorrhoea (4% to 2%). After adjusting for dysmenorrhoea in 1985, each live birth during follow up (OR = 0.20, 95% CI = 0.08 to 0.53) and older age (OR = 0.92, 95% CI = 0.86 to 0.98) were associated with less dysmenorrhoea in 1991.
Conclusions   Primary dysmenorrhoea affects most women throughout the menstrual years. Dysmenorrhoea severe enough to cause absence from work occurs in less than 5% of women. Although improvement and worsening are equally likely for all women, improvement is more likely in women who bear children.     

Fetal growth restriction has long-term effects on postnatal lung structure in sheep

We have previously shown that fetal growth restriction (FGR) during late gestation in sheep affects lung development in the near-term fetus and at 8 wk after birth. In the present study, our aim was to determine the effects of FGR on the structure of the lungs at 2 y after birth; our hypothesis was that changes observed at 8 wk after birth would persist until maturity. FGR was induced in sheep by umbilicoplacental embolization, which was maintained from 120 d until delivery at term (approximately 147 d); birth weights of FGR lambs were 41% lower than in controls. At 2 y after birth, body and lung weights were not different, but there were 28% fewer alveoli and alveoli were significantly larger than in controls; hence there was a 10% reduction in the internal surface area relative to lung volume in FGR sheep compared with controls. The lungs of FGR sheep, compared with controls, had thicker interalveolar septa as a result of increased extracellular matrix deposition; the alveolar blood-air barrier was also thicker, largely because of an 82% increase in basement membrane thickness. These changes are qualitatively similar to those observed at 8 wk. Our data show that structural alterations in the lungs induced by FGR that were apparent at 8 wk were still evident at 2 y after birth, indicating that FGR may result in permanent changes in the structure of the lungs of the offspring and may affect respiratory health and lung aging later in life.