Dendritic cell differentiation pathways of CD34+ cells from the peripheral blood of head and neck cancer patients.

Patients with head and neck squamous cell carcinoma (HNSCC) have increased levels of immune-suppressive peripheral blood CD34+ cells. This study showed that the peripheral blood CD34+ cells of HNSCC patients are capable of differentiating into dendritic cells. Because CD34+ cells can differentiate through several pathways into dendritic cell subpopulations, the intermediate cells through which the blood CD34+ cells of HNSCC patients differentiate were identified. After 6-7 days of culturing the CD34+ cells of HNSCC patients with granulocyte-macrophage colony-stimulating factor, stem cell factor, and tumor necrosis factor at, there appeared CD14+CD1a+ and a lesser proportion of CD14(-)CD1a+ cells resembling the precursor cells of the bipotential and committed dendritic cell differentiation pathways that have been described for cord blood CD34+ cells. To functionally analyze whether these populations were in fact precursor cells, they were isolated and cultured for an additional 10-12 days. Each of these populations was shown to function as precursor cells because they were able to develop into cells that resembled dendritic cells, although a higher proportion developed from the CD14-CD1a+ cells. In contrast, expression of the dendritic activation/maturation marker CD83 was highest on the cells that developed from CD14+CD1a+ cells. Thus, the CD34+ cells whose levels are increased in HNSCC patients can develop into both committed and bipotential dendritic precursor cells, which can subsequently give rise to dendritic cells.

Cultures derived from peripheral blood CD34+ progenitor cells of head and neck cancer patients and from cord blood are functionally different.

Patients with head and neck squamous cell carcinoma (HNSCC) have profound immune defects mediated, in part, by an increased number of immune suppressive CD34+ progenitor cells in their peripheral blood and tumor. One means of overcoming this immune suppression is to stimulate the CD34+ cells to differentiate into more mature, nonsuppressive progeny such as dendritic cells or monocytes. This study determined that CD34+ cells from the peripheral blood of HNSCC patients have the same potential to differentiate into dendritic cells as do human umbilical cord blood CD34+ cells following 12-16 days of culture with a cytokine cocktail. When compared functionally, the cultures that developed from CD34+ cells of cord blood were able to induce an allostimulatory response in naive T-cells, while the cultures that developed from patient CD34+ cells lacked allostimulatory ability. Both cultures expressed class II MHC (HLA-DR), but the proportion of cells expressing the costimulatory molecules CD80 and CD86 was significantly less in cultures that developed from HNSCC-patient CD34+ cells. Therefore, although the CD34+ cells from the peripheral blood of HNSCC patients can differentiate into dendritic cells, their allostimulatory capabilities are impaired, raising the question of their potential effectiveness in stimulating antitumor immune responses.

Molecular markers shared by diverse apomictic Pennisetum species.

Two molecular markers, a RAPD (randomly amplified polymorphic DNA) and a RFLP/STS (restriction fragment length polymorphism/sequence-tagged site), previously were found associated with apomictic reproductive behavior in a backcross population produced to transfer apomixis from Pennisetum squamulatum to pearl millet. The occurrence of these molecular markers in a range of 29 accessions of Pennisetum comprising 11 apomictic and 8 sexual species was investigated. Both markers were specific for apomictic species in Pennisetum. The RFLP/STS marker, UGT 197, was found to be associated with all taxa that displayed apomictic reproductive behavior except those in section Brevivalvula. Neither UGT197 nor the cloned RAPD fragment OPC-04600 hybridized with any sexually reproducing representatives of the genus. The cloned C04600 was associated with 3 of the 11 apomictic species, P. ciliare, P. massaicum, and P. squamulatum. UGT197 was more consistently associated with apomictic reproductive behavior than OPC04600 or cloned C04600, thus it could be inferred that UGT197 is more closely linked to the gene(s) for apomixis than the cloned C04600. The successful use of these probes to survey other Pennisetum species indicates that apomixis is a trait that can be followed across species by using molecular means. This technique of surveying species within a genus will be useful in determining the relative importance of newly isolated markers and may facilitate the identification of the apomixis gene(s).

Transmission of the apomictic mode of reproduction in Pennisetum: co-inheritance of the trait and molecular markers.

Apomixis, asexual reproduction through seed, is an obligate mode of reproduction in several species from the genus Pennisetum. Transfer of apomixis to sexual, cultivated pearl millet (P. glaucum) from a wild species P. squamulatum has resulted in an obligate apomictic backcross line with a low, but unknown number, of chromosomes from the wild species. Molecular markers (restriction fragment length polymorphisms and random amplified polymorphic DNAs) have been identified that unequivocally demonstrate the presence of P. squamulatum DNA in BC3. Three of the informative RFLP clones have been sequenced and converted to sequence-tagged sites that can be amplified by the polymerase chain reaction. Molecular genetic analysis of more advanced back-cross individuals, using the two types of polymerase chain reaction-based molecular markers, has demonstrated co-inheritance of apomictic reproduction and two of the molecular markers. The remaining five molecular markers generally co-segregate with each other but are not linked with the mode of reproduction. These results suggest that genes for apomixis apparently can be transmitted by a single chromosome. Chromosome-specific markers will provide a starting point for the mapping of this genetically intractable reproductive trait.

Postoperative and long-term results of ileal pouch-anal anastomosis for ulcerative colitis and familial polyposis coli.

The immediate postoperative and long-term functional results of 51 ulcerative colitis patients and 21 familial polyposis patients who underwent ileal J-pouch-anal anastomosis were compared in this study. The incidence of postoperative complications requiring reoperation was not statistically different in both groups. The mean daily stool frequency was significantly higher in colitis patients. Pouchitis occurred in 44% of colitis patients but not in polyposis patients (P < 0.005). Symptoms of pouchitis included bloody diarrhea, urgency, abdominal pain, weight loss, fever, and arthritis. Six colitis patients required pouch excision because of intractable pouchitis. The overall pouch excision rate was 22% in ulcerative colitis patients and 5% in familial polyposis patients. Patient satisfaction was good in 46% of ulcerative colitis patients and 76% of polyposis patients (P < 0.05). Our data demonstrate that the long-term outcome of ileal pouch-anal anastomosis is more favorable in polyposis patients than in colitis patients. Pouchitis is a major long-term complication occurring exclusively in colitis patients.

Enterostomy as an adjunct to treatment of intra-abdominal sepsis.

In 60 patients a small bowel enterostomy was constructed as part of the treatment of various intra-abdominal infectious and obstructive conditions. Eleven patients (18 per cent) died in the immediate postoperative period from continuing sepsis. In one patient closure of the stoma was not considered because of disseminated malignancy. In the remaining 48 patients continuity of the gut was subsequently restored. In 22 patients (46 per cent) complications occurred, 12 (25 per cent) of which were intra-abdominal septic complications. The occurrence of intra-abdominal complications was found to be linked to premature (i.e. within 3 months) closure of the stoma. Reasons for premature closure were stomal difficulties and prerenal azotaemia. Stomal closure was attended by a 10 per cent mortality rate.

Mammographic and histopathologic correlation of nonpalpable lesions of the breast and the reliability of frozen section diagnosis.

During a period of ten years, 118 (32.9 per cent) instances of carcinoma were found in 359 specimens taken at biopsy for nonpalpable mammographic lesions. In recent years, the positive predictive value has increased from 68 per cent due to the development of magnification mammography and the use of a mammographic grid. Correlating mammographic and histopathologic data, the rate of malignant disease was 12.7 per cent for instances of a circumscribed or nodular mass, 32.4 per cent for clustered microcalcifications as the only suspect finding, 28.6 per cent when a mass with microcalcifications was present and 66.7 per cent when a stellate-shaped mass was found. Of 188 instances of carcinoma, 40 were noninvasive: 32 instances of ductal carcinoma (27.1 per cent) and eight of lobular carcinoma in situ (6.8 per cent). The possibility of frozen section diagnosis was studied retrospectively by comparison with the paraffin section reports. A correct diagnosis, whether benign or malignant, was achieved in 68 per cent. No frozen section examination was done in 17.3 per cent and the diagnosis was deferred to results of paraffin section in 12.2 per cent. False-negative results were encountered in seven patients (1.9 per cent) and false-positive results in two (0.6 per cent). Both of these patients had florid sclerosing adenosis. Although frozen section diagnosis is feasible in nonpalpable lesions of the breast, it is recommended that this method not be used in instances of pure microcalcifications and tiny solid masses of 5 millimeters or less.

[Misdiagnosed rupture of the diaphragm following blunt thoracic-abdominal trauma]. / Verkannte Zwerchfellrupturen nach stumpfem Brust-Bauch-Trauma.

The histories of 67 patients with a diaphragmatic rupture due to blunt trauma were reviewed in four hospitals. In 45 patients the diagnosis was made within 24 hours after the accident, in the other 22 patients the rupture was diagnosed in a later stage. In the first group there were much abdominal injuries and during emergency laparotomy for some other reasons in 29% of these cases the diaphragmatic rupture was found accidentally. Not recognizing a rupture in the acute phase and therefore delaying operation was caused by the fact that the initial chest X-ray was not thoroughly checked for signs of a diaphragmatic rupture. The reasons for operation in the "delayed" group were mainly typical abnormalities for diaphragmatic rupture of the chest X-rays and other investigations proving the diagnosis. Only in one patient the delay in diagnosis has led to a very serious complication: because of incarceration with gangrene of a part of the small bowel it was necessary to remove this part. The other 21 patients in the group where the diagnosis was initially missed did not suffer from any serious complication.

Localization and excision of nonpalpable breast lesions. A surgical evaluation of three methods.

Three methods of excising nonpalpable breast lesions have been evaluated: (1) "blind" method, using mammographic coordinates; (2) preoperative localization with the Frank needle; and (3) Frank needle localization aided by a multiperforated compression plate. Successful removal at first attempt occurred in about 80% with any method. The size of the biopsy specimens did not differ significantly among the three groups and is most probably a function of the breast volume. The failure rate was seven (2.1%) of 332 biopsies. Since three of the six repeated biopsies yielded specimens with malignancy, the persistence of a radiographically suspicious lesion on follow-up mammogram of the operated-on breast is an urgent indication for reoperation.

Ectopic gastric mucosa in the lower esophagus, presenting as a sessile polyp.

A patient with dysphagia caused by a sessile polyp in the lower esophagus is reported. Histologic examination showed ectopic gastric mucosa. Of the benign tumors of the esophagus, only leiomyoma is seen regularly. The remaining tumors are so rare that the consequences of this diagnosis are unclear. When symptoms make treatment necessary, local excision is preferred.

Perianal fistulas in patients with Crohn's disease.

A series of 55 patients with perianal fistulas and Crohn's disease is reported herein. Thirteen patients (24%) did not need specific treatment of the fistula. Primary proctectomy was necessary in five patients. Defunctioning enterostomy was used in nine patients, followed by healing of the fistula in four patients. Local surgery of the fistula was performed in 28 patients, with an overall success rate of 79%. Healing of the fistula was not influenced by activity of the disease, type of fistula, or condition of the rectum. Local surgery did not cause incontinence in this series.

The significance of microcalcifications without palpable mass in the diagnosis of breast cancer.

Microcalcifications constitute an important part of nonpalpable breast lesions and may be the first sign of a breast carcinoma. Between 1975 and 1984, 150 consecutive patients with clusters of at least five microcalcifications without palpable findings as the only indication for biopsy were treated. One hundred seventy-three groups of microcalcifications were excised and 51 malignancies were detected (29.5%). Most of the malignant lesions were noninfiltrating (56%). Axillary or distant metastases occurred in 11% of the fully evaluable cases. This warrants the expectation that these patients have a very favorable prognosis. Breast biopsy for nonpalpable-clustered microcalcifications is a feasible and valuable procedure. Close cooperation is required between the surgeon, radiologist, and pathologist.

Multicentricity in nonpalpable breast carcinoma and its implications for treatment.

During a 9 year period, 300 consecutive women underwent breast biopsies solely because of nonpalpable, mammographically suspicous findings. One hundred clinically occult breast carcinomas were found, 65 of which were invasive and 35 noninvasive. Eighty-three mastectomy specimens were evaluable for evidence of multifocal carcinoma in another quadrant of the breast or at a distance of 5 cm and residual cancer outside the excisional biopsy cavity. Multicentricity was present in 47 percent and residual tumor in 60 percent of the whole group. When only clinically occult invasive carcinomas were considered, other foci of invasive carcinoma were demonstrated in 26 percent of the patients and residual invasive cancer in 35 percent. The rate of bilaterality was 14 percent, occurring synchronously in 11 percent of the patients. Any therapeutic procedure for nonpalpable breast carcinoma, whether invasive or noninvasive ductal carcinoma, should be directed to the whole breast. Mammography of the contralateral side should be an integral part of the preoperative work-up of patients with palpable lesions ipsilaterally.

Rectal prolapse; a review of Dutch surgical practice.

Many different technics have been described for the treatment of rectal prolapse. Transabdominal fixation of the rectum is the procedure currently most favoured, as in The Netherlands, where the majority of patients with rectal prolapse are treated in this way. A questionaire sent to all teaching hospitals showed that in only seven out of 27 hospitals postanal repair is carried out in cases of postoperative fecal incontinence.

Anastomotic insufficiency in small bowel surgery--incidence and treatment.

In the period 1977-1981 234 small bowel anastomoses were constructed in 143 patients. Eight anastomoses showed leakage (3.4%) and from nine anastomoses a fistula developed (3.8%): a total rate of disturbed healing of small bowel anastomoses (7.3%). In the presence of intra-abdominal infection this rate was 14.8%, in the absence of infection 0.8%. The results of treatment with oversewing and with resection and immediate anastomosis were disappointing. Better results were obtained by dismantling of the anastomosis, establishment of a split-enterostomy and reestablishment of continuity in a second stage. Mortality was 3/17 (18%). The literature is reviewed.