Patterns of reaggregation and formation of linear aggregate chains in collagen-well cultures of dissociated mouse brain and spinal cord cells

With the use of the newly developed collagen-well culture technique spontaneous reaggregation of cells from dissociated embryonic mouse brain and spinal cord were studied. Within 20 h in culture, aggregates are formed and settled onto collagen substrate. Two patterns of aggregate arrangement were observed: random and linear. Linear chains of aggregates appeared to be more characteristic of dissociated spinal cord cells, although the linear patterns were not uncommon in cultures of dissociated cortex. Formation of aggregate chains appeared to be dependent on the stage of neuronal and glial differentiation. After attachment to the collagen substrate, the general pattern of aggregate organization was not greatly altered except for changes which resulted from cellular migration and proliferation, the formation of connections between aggregates, or incorporation of small aggregates into larger ones. The number of non-aggregated cells in collagen-well cultures was small. Single, non-aggregated neurons seldom survived individually. Fiber connections between aggregates began to form after the first day in vitro, and by 2 or 3 days, the growing fibers formed neuritic bridges connecting aggregates. By the end of the first week growing fibers often organized compact bundles, but part connections between aggregates were presented by separate fibers in a diffused manner. Silver impregnation revealed that these connections were formed by the axons of neurons located in the aggregates. Thus, progression of the above described processes resulted in the ‘de novo’ formation of linear organized or random systems of interconnected neuronal centers.     

TP53 codon 72 polymorphism in radiation-associated human papillary thyroid cancer

The study investigated an association between the germline polymorphism at TP53 codon 72 and the development of papillary thyroid cancer (PTC) following exposure to radiation from the Chernobyl accident. TP53 genotype was examined in 48 pediatric/adolescent (age at diagnosis <18 years) and 68 adult post-Chernobyl patient with PTC, 53 adult patients with sporadic PTC and 313 healthy individuals from Russian-Ukrainian population. In addition, we evaluated loss of heterozygosity for TP53 and the allele expression ratio. The genotype of the patients was correlated with clinicopathological data. Arg TP53 homozygotes were found to be significantly underrepresented among adults with post-Chernobyl PTC, but not in children and adolescents when compared with sporadic PTC cases and the general population. In the tumors, cell transformation did not lead to allelic loss or biased TP53 allele expression in heterozygous individuals. None of TP53 genotypes specifically associated with tumor stage and morphology, however there were particular correlations with lymph node status in certain age groups of radiation-associated cases not seen in sporadic PTCs. The findings suggest TP53 allele combinations other than Arg/Arg may contribute to the risk of development of PTC in individuals exposed to radiation during their late childhood, adolescence or in young adulthood.     

Adrenal insuficiency: Diagnosis and management

Adrenal insuficiency is a disorder characterized by hypoactive adrenal glands resulting in insuficient production of the hormones cortisol and aldosterone by the adrenal cortex. This disorder may develop as a primary failure of the adrenal cortex or be secondary to an abnormality of the hypothalamic-pituitary axis. Patients with adrenal insuficiency often are asymptomatic or they may present with fatigue, muscle weakness, weight loss, low blood pressure, and sometimes darkening of the skin. The presentation of adrenal insuficiency varies dramatically and poses a major diagnostic dilemma. This review focuses on the diagnosis and treatment of primary and secondary adrenal insuificency.     

Synthesis and structure-activity relationship of 4-substituted 2-(2-acetyloxyethyl)-8-(morpholine-4-sulfonyl)pyrrolo[3,4-c]quinoline-1,3-diones as potent caspase-3 inhibitors

Synthesis, biological evaluation, and SAR dependencies for a series of novel 1,3-dioxo-2,3-dihydro-1H-pyrrolo[3,4-c]quinoline inhibitors of caspase-3 are described. The inhibitory activity of the synthesized compounds is highly dependent on the nature of 4-substituents on the core scaffold. 4-methyl-and 4-phenyl-substituted derivatives, which were the most active compounds within this series, inhibited caspase-3 with IC50 of 23 and 27 nM, respectively.     

Pelvic floor disorders: linking genetic risk factors to biochemical changes

Pelvic floor disorders (PFDs) such as stress urinary incontinence (SUI) and pelvic organ prolapse (POP) may share a common pathophysiological process related to pelvic floor tissue laxity and loss of support. We reviewed recent literature on observed biochemical changes in women with SUI and POP, linking them to genetic predisposition. We found that studies of pelvic tissues showed differences between control subjects and women with POP and SUI in collagen and elastin structure at a molecular and fibrillar level. Studies were heterogeneous but showed a trend towards decreased collagen and elastin content. The contribution of matrix metalloproteinases to increased collagenolysis can be related to genetic polymorphisms present in higher frequency in women with PFD. Extracellular matrix (ECM) protein turnover plays a role in the development of POP and SUI, but much remains to be understood of this complex dynamic interplay of enzymes, proteins and molecules. Genotyping of candidate genes participating in ECM formation will elucidate the missing link between the manifestation of the disease and the biochemical changes observed systematically, in addition to those in the pelvic floor.     

Neck injury and fibromyalgia-- are they really associated?

OBJECTIVE: To investigate whether whiplash injury may be a trigger for the onset of fibromyalgia (FM). METHODS: One hundred fifty-three patients presenting to the emergency room with the diagnosis of whiplash injury were examined. The control group included 53 patients hospitalized with fractures of the limbs, spine, and ribs due to road accident. The study and control groups were interviewed shortly after presenting and then followed prospectively. Patients complaining of musculoskeletal symptoms during followup were examined and a count of 18 tender points was conducted. FM was diagnosed if the patient fulfilled currently accepted 1990 American College of Rheumatology criteria. RESULTS: The mean followup period for the study and control groups was 14.5 months (range 12-18) and 9 months (range 6-14), respectively. There were no differences between the groups with regard to age, sex, marital, education, or employment status. During the followup period only one patient in the study group and no patients in the control group developed signs and symptoms of FM. Three patients in the study group (2%) and 15 patients in the control group (16%) filed insurance claims; none was associated with FM. CONCLUSION: Whiplash injury and road accident trauma were not associated with an increased rate of FM after more than 14.5 months of followup.