Reflection of neuroblastoma intratumor heterogeneity in the new OHC-NB1 disease model

Accurate modeling of intratumor heterogeneity presents a bottleneck against drug testing. Flexibility in a preclinical platform is also desirable to support assessment of different endpoints. We established the model system, OHC-NB1, from a bone marrow metastasis from a patient diagnosed with MYCN-amplified neuroblastoma and performed whole-exome sequencing on the source metastasis and the different models and passages during model development (monolayer cell line, 3D spheroid culture and subcutaneous xenograft tumors propagated in mice). OHC-NB1 harbors a MYCN amplification in double minutes, 1p deletion, 17q gain and diploid karyotype, which persisted in all models. A total of 80–540 single-nucleotide variants (SNVs) was detected in each sample, and comparisons between the source metastasis and models identified 34 of 80 somatic SNVs to be propagated in the models. Clonal reconstruction using the combined copy number and SNV data revealed marked clonal heterogeneity in the originating metastasis, with four clones being reflected in the model systems. The set of OHC-NB1 models represents 43% of somatic SNVs and 23% of the cellularity in the originating metastasis with varying clonal compositions, indicating that heterogeneity is partially preserved in our model system.     

Inhibition of cellular immune reactions by zymosan

Mice treated intraperitoneally with zymosan showed a strong inhibition of the DTHR (delayed-type hypersensitivity reaction) against sheep erythrocytes (SE), ovalbumin (OA), and alloantigen. Furthermore, the rejection time of skin transplants was nearly doubled while antibody formation against SE was significantly enhanced. When a DTHR against OA and an antibody formation against SE were induced at the same time in the same animals, than the suppressive and stimulating effects cancelled each other. These results are discussed with regard to the sensitivity of lymphocyte subpopulations, which may be different if exposed to phagocytosis-induced oxygen radicals.     

Reflex sympathetic dystrophy in cervical spinal cord injury patients

Sixty consecutive patients admitted to the spinal cord injury unit at a Downey, California medical center were evaluated for hand and upper extremity pain. Patients averaged nine months postinjury and had an average age of 37 years. Seven patients (11.7%) complained of diffuse hand pain, swelling, and stiffness. All patients with complaints were evaluated with three-phase radionuclide scintigraphy. Six of those seven patients had scintigrams consistent with reflex sympathetic dystrophy (RSD), an overall incidence of 10%. Three of these six patients were treated with stellate ganglion blocks, which gave relief of symptoms and allowed return to their rehabilitation program. An awareness of RSD as a cause of pain in spinal cord injured patients should lead to earlier recognition and treatment.     

Atypical presentation of co-existent Haemophilus ducreyi and Treponema pallidum infection in an HIV-positive male.

A 25-year-old homosexual black male presented with asymmetrical perianal ulceration of uncertain clinical origin. Indepth microbiological examination revealed the combined presence of Haemophilus ducreyi and Treponema pallidum. The atypical clinical appearance may have been due to the changed immunological status of the host's being infected with Human Immunodeficiency Virus.     

Sequence of the human 40-kDa keratin reveals an unusual structure with very high sequence identity to the corresponding bovine keratin.

The complete amino acid and DNA sequences of the human 40-kDa keratin are reported. The DNA sequence encodes a protein of 44,098 Da, which is unique in that it lacks the terminal non-alpha-helical tail segment found in all other keratins. When the human 40-kDa keratin amino acid sequence is compared to the corresponding bovine keratin, the overall identity is 89%. The coil-forming regions are 89% identical and the head regions are 88% identical. This similarity is also evident in the DNA sequence of the coding region, the 5' upstream sequences, and the 3' noncoding sequences. The high degree of cross-species identity between bovine and human 40-kDa keratins suggests that there is strong evolutionary pressure to conserve the structure of this keratin. This in turn suggests an important and universal role for this intermediate filament subunit in all species.     

Maternal transmission of immunity to Eimeria maxima: western blot analysis of protective antibodies induced by infection.

Infection of breeding hens with Eimeria maxima induces production of parasite-specific antibodies which are transferred, via the egg yolk, to hatchling chicks. These antibodies (immunoglobulin G) are highly protective, mediating up to a 97% reduction in oocyst excretion in challenged hatchlings. However, the degree of maternally derived immunity transferred by the hens to their offspring declines with increasing time after infection of the hens. This decline in immunity is directly related to declining immunoglobulin G titers. However, sera from highly protected hatchlings recognize only a very few E. maxima proteins on Western blots (immunoblots). In particular, a 230-kDa protein band is outstanding for its association with maternally derived immunity to E. maxima in hatchlings. This band was excised from a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) preparative gel of crude merozoite protein extract. The SDS-PAGE cutout was emulsified in Freund's adjuvant and injected, intramuscularly, into six breeding hens on two occasions, 2 weeks apart. Eggs were collected from these hens 28 to 39 days after the second injection, and the hatchlings from these eggs were challenged with 150 sporulated oocysts of E. maxima. Subsequent oocyst excretion in these hatchlings was, on average, 54% lower than oocyst excretion by control chicks but only 37% lower (significant at P < 0.05) than that by chicks from hens sham immunized with Freund's adjuvant. The latter result is apparently due to the ability of the adjuvant to induce production of antibodies which recognize Eimeria spp. and thereby transfer some degree of protection to hatchlings. These experiments indicate that protective, maternally derived immunoglobulin G antibodies may be useful for the identification of putative anticoccidial vaccine candidates.     

Naevus follicularis keratosus: clinical aspects, histology and histogenesis]

The clinical, histological and histogenetic aspects of naevus follicularis keratosus (NFK) ("naevus comedonicus") are reported. Clinically, NFK appears mostly as linear and unilateral groups of dark comedo-like plugs. Clinical forms include variants with minimal and distinctive deviations from the basic form. Recurrent inflammation is not mandatory. Histological examination reveals keratin-filled infundibula, with granular layers that are always present but though not always equally obvious. This finding corresponds to the mode of keratinization in the follicular infundibulum. Overall, the findings are indicative of a harmartoma of the follicular infundibulum with additional rudimentary sebaceous glands.