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Group singing and playing of wind instruments increase COVID-19 transmission risk. After a pause during the initial period of the COVID-19 pandemic, The Tabernacle Choir at Temple Square organization (hereinafter, Choir) resumed musical events in September 2021 with prevention protocols, including required vaccination and pre-event rapid antigen testing. We investigated potential SARS-CoV-2 transmission at Choir events during September 21-November 7, 2021. We interviewed COVID-19-positive members (hereinafter, case-members) and identified members exposed when a case-member attended a Choir event during his or her infectious period. We compared whole genome sequencing results to assess the genetic relatedness of available SARS-CoV-2 specimens obtained from case-members. We identified 30 case-members through pre-event testing (n = 10), self-reported positive test results (n = 18), and a review of Utah's disease surveillance system (n = 2). All 30 case-members reported symptoms; 21 (70%) were women and 23 (77%) received a positive test result by nucleic acid amplification test. No hospitalizations or deaths were reported. We identified 176 test-eligible exposed members from 14 instances of case-members attending events during their infectious periods. All were tested at least once 2 to 14 days after exposure: 74 (42%) by rapid antigen test only (all negative) and 102 (58%) by nucleic acid amplification test (4 positive, 97 negative, and 1 equivocal). Among viral sequences available from 15 case-members, the smallest single-nucleotide polymorphism distance between 2 sequences was 2, and the next-smallest distance was 10. The lack of disease detected in most exposed members suggests that minimal, if any, transmission occurred at Choir events. When community COVID-19 incidence is high, prevention protocols might help limit SARS-CoV-2 transmission during group musical activities.

Young Black gay, bisexual, and other men who have sex with men (YB-GBMSM) living with HIV are at high risk for psychological stressors and comorbidities. We sought to gain the perspectives of multidisciplinary providers (including HIV care, mental health care, and social service providers) on their experiences with engaging YB-GBMSM in mental health care within comprehensive HIV primary care centers that included both types of services on-site. We conducted qualitative in-depth interviews with fifteen providers in two clinics, and analyzed the qualitative data using a thematic analysis approach. Our participants described high levels of need for mental health services among YB-GBMSM living with HIV, due to psychological challenges related to living with HIV as well as other frequently occurring life stressors. At the same time, barriers to accessing these services were identified, with mental health stigma, under-recognition of symptoms and confidentiality concerns being the most commonly cited. Strategies to improve mental health service uptake in light of these needs and barriers were discussed, including re-framing of mental health services, addressing logistical barriers, and utilization of a biopsychosocial perspective.

Young Black gay, bisexual and other men who have sex with men (YB-GBMSM) are disproportionately burdened by HIV and often exhibit suboptimal engagement in HIV care. With the goal of increasing engagement in HIV care, we designed a culturally specific, theory-based group-level program, Brothers Building Brothers by Breaking Barriers (B6), which aimed to strengthen resilience and social capital among YB-GBMSM living with HIV. We conducted a pilot trial to evaluate the program's acceptability and feasibility. Through clinic-based recruitment and community outreach events, we recruited and enrolled 71 YB-GBMSM into the study. Participants were randomized to either the B6 program or a control comparison program. Post-session evaluation surveys and in-depth qualitative interviews showed B6 to have high levels of acceptability and satisfaction. Specifically, participants described benefits to interacting in a group with other YB-GBMSM, and several described increased comfort with their own gay identities after participation. No adverse events or safety concerns were reported. However, there were challenges to feasibility, as reflected in recruitment and retention rates. The B6 program was highly acceptable among YB-GBMSM living with HIV; however, innovative program delivery methods and implementation strategies will be needed to improve recruitment and retention in future implementation of B6., (© The Author(s) 2022. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

OBJECTIVE The aim of this study is to describe a technique for percutaneous cervical transfacet screw placement and compare this technique to the open technique with regard to the accuracy of facet capture and the potential of placing neurovascular structures at risk. METHODS Eight cadaveric cervical spines were harvested. One side of each spine was assigned to the percutaneous group, and the other side to the open group. The spines were instrumented from C-3 to T-1 (80 screws). The distance to the spinal canal, foramen transversarium, and neural foramen were measured to determine the likelihood of placing neurovascular structures at risk. The percentage of the facet joint captured and the angle of screw trajectory compared with the ideal trajectory were used to determine the accuracy. RESULTS There were, in total, 11 misplacements of screws: 2 screws using the open technique and 9 screws using the percutaneous technique (p = 0.006). From a neurovascular point of view, 3 percutaneous screws violated the foramen transversarium. Two of these percutaneous screws violated the neural foramen. No neurovascular foramina were violated using the open technique. The open technique resulted in a significantly greater distance from the screw to the spinal canal (p < 0.001). The distance from the screw to the foramen transversarium (p = 0.015), as well as the distance from the screw to the neural foramen (p = 0.012), did not demonstrate statistical difference when using either technique. As for the accuracy of facet capture, 8 screws exhibited less than 15% purchase of the facet joint. Six of these screws were placed using the percutaneous technique, and 2 screws were placed using the open technique. CONCLUSIONS There is a higher incidence of screw misplacement using the percutaneous transfacet in comparison to the open transfacet technique. The accuracies of facet capture using the 2 techniques were not statistically different. Surgeons will need to understand the potential risk of using the percutaneous technique as an alternative to open transfacet fixation.

Background: Screw length pertains to stability in various orthopedic fixation devices. There is little or no information on the relationship between plate pullout strength and screw length in anterior lumbar interbody fusion (ALIF) plate constructs in the literature. Such a description may prove useful, especially in the treatment of osteoporotic patients where maximizing construct stability is of utmost importance. Our purpose is to describe the influence of screw length on ALIF plate stability in severely and mildly osteoporotic bone foam models., Methods: Testing was performed on polyurethane foam blocks with densities of 0.08 g/cm(3) and 0.16 g/cm(3). Four-screw, single-level ALIF plate constructs were secured to the polyurethane foam blocks by use of sets of self-tapping cancellous bone screws that were 20, 24, 28, 32, and 36 mm in length and 6.0 mm in diameter. Plates were pulled out at 1 mm/min to failure, as defined by consistently decreasing load despite increasing displacement., Results: Pullout loads in 0.08-g/cm(3) foam for 20-, 24-, 28-, 32-, and 36-mm screws averaged 303, 388, 479, 586, and 708 N, respectively, increasing at a mean of 25.2 N/mm. In 0.16-g/cm(3) foam, pullout loads for 20-, 24-, 28-, 32-, and 36-mm screws averaged 1004, 1335, 1569, 1907, and 2162 N, respectively, increasing at a mean of 72.2 N/mm., Conclusions: The use of longer screws in ALIF plate installation is expected to increase construct stability. Stabilization from screw length in osteoporotic patients, however, is limited.

Background Context: Although the frequency of transpsoas lumbar interbody fusion procedures has increased in recent years, complication reports remain scarce in the literature., Purpose: To present four cases of vertebral body fracture after transpsoas interbody fusion procedures in nonosteoporotic patients without significant trauma and discuss relevant biomechanical factors., Study Design: Case series and literature review., Patient Sample: Patients 1 and 2 were obese men who underwent one- and two-level transpsoas interbody fusion procedures and subsequently experienced coronal plane fracture. Patients 3 and 4 were elderly women who underwent multilevel transpsoas interbody fusion procedures and experienced L5 compression fracture., Results: Patients 2 and 3 were treated nonsurgically after fracture. The fractures healed uneventfully; however, Patient 3 developed a flat back syndrome. Patient 1 underwent posterior instrumented fusion and had solid bridging bone above and below the fracture. Patient 4 was treated with vertebroplasty. Factors potentially contributing to these fractures were discussed., Conclusions: Fracture can occur after transpsoas lumbar interbody fusion, even in nonosteoporotic patients. Factors, such as intraoperative end-plate breach, subsidence, compression by lateral screws, and cage rolling, could contribute to the development of fractures after transpsoas interbody fusion., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Study Design: Retrospective magnetic resonance imaging (MRI)-based study., Objective: Our goal was to develop Wiltse's paraspinal surgical approach by determining the precise anatomic locations of the intermuscular cleavage planes formed by the multifidus and longissimus muscles. The primary objective was to measure the distances between the midline and the intermuscular planes, bilaterally, on MRI scans at each of the five disc levels between L1 and S1. Secondary objectives included identifying the existence of any correlations between patient demographics and the measured outcomes., Summary of Background Data: In 1968, Wiltse described an approach to the spine using the natural cleavage plane of the multifidus and longissimus muscles as an entry to the posterior spinal elements. The small direct incisions lessened bleeding, tissue violation, and muscle retraction, which popularized Wiltse's approach among surgeons. A detailed description of the locations of the intermuscular cleavage planes at each lumbar disc level, however, is not available., Methods: MRI scans of 200 patients taken during routine care (2007-2009) were retrospectively reviewed to gather measurements of the distances from the intermuscular cleavage planes to the midline, bilaterally, at each disc level from L1 to S1. Age, sex, and BMI (body mass index) were obtained to determine correlations., Results: Mean measurements significantly differed between all disc levels. At L5-S1, the mean distance was 37.8 mm; at L4-L5, 28.4 mm; at L3-L4, 16.2 mm; at L2-L3, 10.4 mm; and at L1-L2, 7.9 mm. The mean female distances were significantly greater than males (2 mm) on both sides of L5-S1 only. No correlation was discovered between BMI, age, height (N = 50), or weight (N = 50) with respect to measured distances., Conclusion: In the absence of any significant clinical correlation between patient demographics and the entry site in Wiltse's approach, the spine surgeon may use distances described in this paper to apply to a broad base of spine patients regardless of BMI, sex, or age.

Background: The increase in total disc replacement procedures performed over the last 5 years has increased the occurrence of patients presenting with postoperative iatrogenic deformity requiring revision surgery. Proposed salvage treatments include device retrieval followed by anterior lumbar interbody fusion or posterior fusion. We propose a novel approach for the correction of disc tilt after total disc replacement using a posterior dynamic stabilization system., Methods: Pedicle screws can be inserted either in an open manner or percutaneously by standard techniques under fluoroscopy. The collapsed side is expanded, and the convex side is compressed. Universal spacers are placed bilaterally, with the spacer on the collapsed side being taller by 6 mm. Cords are threaded through the spacers and pulled into place with the tensioning instrument. Extra tension is applied to the convex side, and the wound is closed by standard techniques., Results: Three patients presenting with tilted total disc replacement devices underwent corrective surgery with posterior dynamic stabilization. Radiographs confirmed correction of deformity in all cases., Conclusions/level of Evidence: This technical note presents a novel indication for posterior dynamic stabilization and describes its surgical application to the correction of disc tilt after total disc replacement. This is level V evidence.

Total disc replacement (TDR) surgeries have significantly increased in number since US FDA approval. However, the associated complications such as implant malposition, loosening, subsidence, implant migration, fractures, and infection may necessitate retrieval of the artificial disc and proceeding with interbody fusion. Retrieval of artificial discs in proximity to important vascular, urological, and other vital anatomical structures (for example, L4-5) presents numerous difficulties to spine and approach surgeons. To avoid the impediments of an anterior revision approach, in 2006 Pimenta et al. described an effective transpsoas lateral minimally invasive approach to retrieve the TDR device. In this paper, the authors adopted this technique in their surgical practice; however, they realized that a complex step involved the removal of the polyethylene core. The polyethylene core is compressed between the device endplates and is difficult to remove with the aid of a Kocher clamp as advised by Pimenta et al. Moreover, distraction on the endplates or the vertebral bodies to decrease the compression across the core is laborious, time consuming, and often not possible due to the approach and location of the lumbar plexus. In the present paper, the authors propose a novel modification to the polyethylene core removal with the use of a screw to create a better lever arm, apply effective distraction, and secure a good grip to enable core retrieval. This modification significantly reduced the operating time.

MM14 murine myoblast cells remain in an undifferentiated and proliferative state if they are maintained in the continuous presence of basic fibroblast growth factor (FGF-2) and serum factors, but terminally differentiate into myocytes and myotubes if deprived of FGF-2 during G1 of the cell cycle. We find that 2-aminopurine (2-AP) induces a reversible, rapid, and profound alteration in the morphology of proliferating MM14 cells to a polarized shape which is similar to that observed during normally induced differentiation. This change requires neither a differentiation signal nor de novo protein synthesis. In contrast, we do not observe a morphological change in response to 2-AP in nonmyogenic cell lines. The morphological alteration of MM14 cells in response to 2-AP requires reorganization of the microtubule and actin cytoskeletons, in common with changes during normal differentiation. Additionally, we show that cytoskeletal rearrangements that occur during both normal differentiation and in response to 2-AP require the activity of the small GTPase Rho. Differentiation defective MM14 cells (DD-1 cells), which lack MyoD, also undergo a profound morphological alteration in the presence of 2-AP. These results indicate that morphological changes consistent with myoblast differentiation are regulated by a pathway independent of MyoD transcriptional control.

Thymic stromal cell line TS-9 was found to selectively bind a subpopulation of normal murine thymocytes. Selective binding allowed the isolation and phenotypic characterization of the adherent and nonadherent subpopulations of thymocytes. Flow cytometric analysis of fluorescently labeled thymocytes revealed that the adherent and nonadherent populations differ in maturity, with the adherent population enriched in immature thymocytes of the PNAhi, Thy-1hi, CD3-/lo, and CD4+/CD8+ double positive surface phenotype. A quantitative microwell assay was developed to measure the binding of thymocytes to TS-9. Thymocytes labeled with vital DNA stain Hoechst 33342 were allowed to bind to TS-9 in microwells and the intense fluorescence of this label was readily detected with a scanning fluorometer. The binding was trypsin-sensitive and hyaluronidase and PI-PLC resistant. The binding was also temperature dependent and sensitive to cytochalasin B. A panel of monoclonal antibodies to cell surface antigens including CD2, LFA-I/ICAM-I, and Thy-1 was screened in a quantitative binding assay for their ability to inhibit the binding of thymocytes to TS-9. The binding was partially inhibited by the C3C12 monoclonal antibody which recognizes the recently identified and apparently unique gp23,gp45 complex expressed on murine stromal cells.

We have discovered a novel mitosis-specific human autoantigen that arises at the centromeres of prophase chromosomes, but ultimately participates in formation of an organelle that bisects the cell at late anaphase and during telophase. The organelle, discernible as a three-dimensional disc by confocal microscopy, encompasses the entire midzone diameter, and its distribution survives disassembly of interpolar microtubules by cold temperature treatment and detergent lysis of cells. Cytokinetic furrow contraction proceeds normally in dihydrocytochalasin B (DCB)-treated cells, and antigen distribution in the furrow is unaltered. In DCB, the furrow retracts in early interphase, coincident with loss of normal membrane association with the disc, resulting in the formation of binucleate cells. The midzone disc in both drug-treated and normal cells is present at the correct time and position to play a central role in cytokinesis. By immunocytochemistry, the disc appears to contain myosin but not actin. The position of the disc and the possible presence of myosin suggest that cytokinesis may involve the interaction of the disc organelle with actin in the cell cortex to produce cleavage in mammalian cells.

CENP-A, a centromere-specific 17-kDa protein, has histone-like properties. However, in contrast to the common somatic histones, CENP-A is quantitatively retained in bull spermatozoa, and we have exploited this fact to purify CENP-A to apparent homogeneity. Partial sequence analysis of the purified protein indicates that CENP-A is a distinctive gene product. Some CENP-A sequences are highly similar to regions of histone H3. Other segments of CENP-A are not related to H3 or any other histone. These unrelated segments are presumably involved in localizing CENP-A to centromeric DNA or in centromere-specific functions of CENP-A.

The 17 kDa human autoantigen designated CENP-A is a centromere specific histone. We show here that CENP-A is present in tissue of bovine origin, and that it is quantitatively retained in mature spermatozoa. This result is striking, as a prominent feature of spermatogenesis in mammals is the replacement of most somatic and testes specific histones with protamines. Indirect immunofluorescence studies further show that CENP-A is retained in sperm nuclei in discrete foci, rather than being dispersed throughout the sperm head. These observations suggest that CENP-A is a functionally important component of centromeres, and that pre-existing CENP-A:DNA interactions are likely to be important in organizing the centromeres of the paternal genome during early embryogenesis.

The events of mammalian fertilization overlap with the completion of meiosis and first mitosis; the pro-nuclei never fuse, instead the parental genomes first intermix at the mitotic spindle equator at metaphase. Since kinetochores are essential for the attachment of chromosomes to spindle microtubules, this study explores their appearance and behavior in mouse oocytes, zygotes and embryos undergoing the completion of meiosis, fertilization and mitoses. Kinetochores are traced with immunofluorescence microscopy using autoimmune sera from patients with CREST (CREST = calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, telangiectasia) scleroderma. These sera cross-react with the 17 kDa centromere protein (CENP-A) and the 80 kDa centromere protein (CENP-B) found at the kinetochores in human cell cultures. The unfertilized oocyte is ovulated arrested at second meiotic metaphase and kinetochores are detectable as paired structures aligned at the spindle equator. At meiotic anaphase, the kinetochores separate and remain aligned at the distal sides of the chromosomes until telophase, when their alignment perpendicular to the spindle axis is lost. The female pronucleus and the second polar body nucleus each receive a detectable complement of kinetochores. Mature sperm have neither detectable centrosomes nor detectable kinetochores, and shortly after sperm incorporation kinetochores become detectable in the decondensing male pronucleus. In pronuclei, the kinetochores are initially distributed randomly and later found in apposition with nucleoli. At mitosis, the kinetochores behave in a pattern similar to that observed at meiosis or mitosis in somatic cells: irregular distribution at prophase, alignment at metaphase, separation at anaphase and redistribution at telophase. They are also detectable in later stage embryos. Colcemid treatment disrupts the meiotic spindle and results in the dispersion of the meiotic chromosomes along the oocyte cortex; the chromosomes remain condensed with detectable kinetochores. Fertilization of Colcemid-treated oocytes results in the incorporation of a sperm which is unable to decondense into a male pronucleus. Remarkably kinetochores become detectable at 5 h post-insemination, suggesting that the emergence of the paternal kinetochores is not strictly dependent on male pronuclear decondensation.

A monoclonal antibody, 1D8, which recognizes a cell-surface antigen expressed by human chromosome 3 in Chinese hamster-human somatic-cell hybrids, has been produced. Testing of hybrids containing various deletions of chromosome 3 determines that the gene encoding the antigen is regionally localized to 3q (cen-22). This regional mapping is distinct from that elsewhere reported for two other cell-surface antigens assigned to chromosome 3--namely, the human transferrin receptor and the p97 melanoma-associated antigen. In addition, biochemical characterization is different from that elsewhere reported for other chromosome 3-encoded cell-surface antigens. When tested against a panel of rare-phenotype red blood cells, the only cells that failed to react were those of the Rhnull phenotype. The antibody reacts only weakly with homozygous -D- and fetal red cells, in contrast with a previously described antibody, R6A, which does not react with Rhnull cells. Furthermore, R6A does not recognize a cell-surface antigen expressed by chromosome 3 in Chinese hamster-human somatic-cell hybrids. Thus, the monoclonal antibody 1D8 recognizes a previously undescribed cell-surface antigen encoded by human chromosome 3 and not expressed on Rhnull cells. The gene on chromosome 3 regulating expression of this antigen may be that defective in Rhnull disease or may require the normal allele at an unlinked Rhnull locus for expression. Linkage studies will be required to further elucidate this matter.

Chicken phosphoglucomutase (PGM-2), serum albumin, vitamin D binding protein (Gc) and phosphoribosyl pyrophosphate amidotransferase (PPAT) structural genes have been mapped to chicken chromosome 6 using chicken-Chinese hamster somatic cell hybrids containing this chromosome as the only chicken genetic material. Chicken PGM-2 activity was detected in the hybrids using cellogel electrophoresis and a substrate, ribose-1-phosphate (R-1-P), that allows the detection of PGM-1 activity in mice and PGM-2 activity in humans. Chicken albumin sequences were detected in the hybrids with the use of a labelled chicken serum albumin cloned cDNA. Classical studies have shown linkage of the serum albumin and Gc genes, and the Gc gene also can be localized to chicken chromosome 6. The PPAT gene was localized to this chromosome in previous studies using these hybrids. A homologous linkage group has been identified in mammals and, therefore, a chromosomal linkage group containing at least four genes--Gc, serum albumin, PPAT, and PGM-2--has been conserved over a period of 300 million years, throughout both avian and mammalian evolution.

An attempt was made to identify erythrocyte cell surface antigens associated with the major histocompatibility complex in the turkey. Two turkey lines, A and B, from the University of Minnesota, putatively homozygous for different major histocompatibility complex alleles, were used for this study. A total of 79 birds were immunized either by skin grafting or by injection of homogenized skin preparation in Freund's incomplete adjuvant. Antisera produced with skin grafts and homogenized skin inoculated intramuscularly both contained red cell agglutinins of low titer. Because agglutination reactions were weak, no sera were suitable for use as line-specific typing fluids. Thus, we were unable to demonstrate that turkey erythrocytes contain serologically detectable major histocompatibility antigens corresponding to the B major histocompatibility system in the chicken or the H-2 system in mice. Rather, the turkey major histocompatibility complex is more like that in rabbits, dogs, monkeys and humans.

Two murine monoclonal antibodies, 1D12 and 2F7, apparently of the same specificity, define a new red cell polymorphism, MER2. 92% of English blood donors are MER2+ giving the gene frequencies MER2+ 0.7159; MER2- 0.2841. Family studies showed that MER2+ is inherited as a Mendelian dominant character and that MER2 is not controlled by any of the main blood group loci. The MER2 antigen is also present on some leukaemia fibroblasts and human cell lines. Using somatic cell hybrids, MER2 appears to be coded for by a gene on chromosome 11 at 11p15.

Five monoclonal antibodies which indirectly agglutinate human rbc were isolated and designated 2D8, 4E12, 4H10, 5A8, and 5H5. Expression of the antigen defined by 2D8 was 100% concordant with the presence of human chromosome 19 in a panel of human-Chinese hamster somatic cell hybrids. Secondary clones isolated from antigen positive hybrids exposed to antibody 2D8 and complement were shown to have lost both the cell surface antigen and chromosome 19. Small terminal deletions of 19p were observed in the karyotypes of two antigen negative clones that continued to express human GPI. All five monoclonal antibodies and the previously isolated F10 labelled the surface of parental cells and did not label the surface of deletion mutants, and the five monoclonal antibodies produced in this study inhibited the binding of F10 to the human rbc surface. Therefore, all must recognize the MSK20 cell surface antigen which was defined previously by antibody F10. Chimpanzee cells bind 5A8 and 5H5, orangutan cells bind 4E12, and neither bind 2D8 and 4H10. Therefore, the monoclonal antibodies produced in this study must recognize at least three distinct epitopes of the MSK20 cell surface antigen which are expressed on the human rbc.

DNA-mediated transfectants were isolated that expressed two of the cell-surface antigens encoded by human chromosome 11. These tranfectants were used to analyze monoclonal antibodies selected to recognize human cell-surface antigens expressed by a somatic cell hybrid containing 11 as its only human chromosome. Analysis of the transfectants, deletion hybrids, and mutants showed that the monoclonal antibodies recognized at least five different antigens, one of which we had not identified previously. A majority of the monoclonal antibodies recognized the a1 antigen. The use of cells from higher primates demonstrated that the a1-specific monoclonal antibodies recognize at least two epitopes.

Drosophila chromatin contains two antigenically distinct H2A histones, H2A.1 and H2A.2. Indirect immunofluorescence analyses revealed that anti-H2A.1 binding was distributed throughout polytene chromosomes, whereas anti-H2A.2 binding was interband-specific. Thus, H2A.2 probably contributes to the less compacted structure of interbands. Since each band-interband region is thought to contain a single gene, our results suggest that the distribution of H2A.2 echoes the functional organization of the Drosophila genome. Similar H2A histones occur in eukaryotes ranging from protozoa to mammals. Their placement might be an important determinant of chromatin structure.

Hybridomas producing monoclonal antibody (MAb) of predefined specificity were isolated from a Chinese hamster that had received injections of a Chinese hamster-human somatic cell hybrid. A standard method for the production of murine hybridomas was used to produce Chinese spleen lymphocyte X murine plasmacytoma hybridomas, and they were screened for complement-mediated cytotoxicity against a panel of Chinese hamster-human somatic cell hybrids and agglutination of human erythrocytes. Two hybridomas were established in tissue culture following limiting dilution cloning, and their reactivity with a panel of Chinese hamster-human somatic cell hybrids indicates that they are specific for the previously identified human cell-surface antigen a1. The Chinese hamster appears to respond preferentially to human antigens of Chinese hamster-human somatic cell hybrids, and it will serve as a useful tool for the production of MAb specific for human cell-surface antigens expressed in the many well-characterized Chinese hamster-human somatic cell hybrids available.

In embryonic nuclei of Drosophila virilis, 45% of the DNA is satellite, and congruent to 50% of the H1 histone is phosphorylated. In polytene salivary gland nuclei, less than 1% of the DNA is satellite, and less than 10tion. The phosphorylated H1's migrate 4% slower than the unphosphorylated H1's on SDS-acrylamide gels. The mobility difference may arise because the phosphorylated and unphosphorylated H1's have different conformations in SDS. This putative conformational difference could be essential to the compaction of satellite DNA into heterochromatin.

We have developed a competitive enzyme-linked immunosorbent assay for solubilized kinetochore components, using human CREST (calcinosis, Raynaud's phenomenon, esophageal dysfunction, sclerodactyly, telangiectasia) scleroderma autoimmune antibodies specific for these kinetochore elements. Using this quantitative assay, we found interphase persistent or "pre-kinetochore" components in low- and moderately high-salt (375 mM salt) extracts of micrococcal nuclease-digested rat liver and chicken erythrocyte nuclei. The release of antigen activity from nuclei under these conditions has been correlated with loss of pre-kinetochore foci as determined by immunofluorescence microscopy. Combined biochemical and competition assay analysis of chicken erythrocyte nuclear extracts indicates that pre-kinetochore components are tightly bound to chromatin of mononucleosome size. The conclusions based on competition assay data are supported by a direct binding assay, which confirms that antigens recognized by CREST sera are present on chromatin. These results raise the possibility that the kinetochore-specific chromosomal antigen(s) we have detected substitutes for "standard" mononucleosome components, such as histone H1. Furthermore, they suggest approaches to the isolation of kinetochore-specific DNA sequences from higher eucaryotes.