Principles and analytical performance of Papilloplex® HR-HPV, a new commercial CE-IVD molecular diagnostic test for the detection of high-risk HPV genotypes.

The accurate detection and genotyping of high-risk human papillomavirus (HR-HPV) are critical for cervical cancer screening and epidemiological investigations. GeneFirst Papilloplex® HR-HPV is a new CE-IVD-marked real-time PCR test based on patented multiplex probe amplification technology. Papilloplex® HR-HPV provides the simultaneous detection and differentiation of 14 HR-HPV genotypes (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68a/b) in a single closed-tube reaction ensuring rapid, cost-effective, and contamination-free results. In this study, the analytical performance characteristics in terms of the assay's sensitivity, specificity, range, reproducibility, and cross-reactivity were evaluated. Papilloplex® HR-HPV provided sensitive detection and differentiation of 14 HR-HPV types with highly reproducible results. The differential HR-HPV specificity and sensitivity were further confirmed through the participation in the WHO HPV Laboratory Network Proficiency Study (2014). Overall, GeneFirst Papilloplex® HR-HPV assay demonstrated a robust analytical performance with reproducible and reliable results in the detection of HR-HPV genotypes.

Genetic and functional analyses identify DISC1 as a novel callosal agenesis candidate gene.

Agenesis of the corpus callosum (AgCC) is a congenital brain malformation that occurs in approximately 1:1,000-1:6,000 births. Several syndromes associated with AgCC have been traced to single gene mutations; however, the majority of AgCC causes remain unidentified. We investigated a mother and two children who all shared complete AgCC and a chromosomal deletion at 1q42. We fine mapped this deletion and show that it includes Disrupted-in-Schizophrenia 1 (DISC1), a gene implicated in schizophrenia and other psychiatric disorders. Furthermore, we report a de novo chromosomal deletion at 1q42.13 to q44, which includes DISC1, in another individual with AgCC. We resequenced DISC1 in a cohort of 144 well-characterized AgCC individuals and identified 20 sequence changes, of which 4 are rare potentially pathogenic variants. Two of these variants were undetected in 768 control chromosomes. One of these is a splice site mutation at the 5' boundary of exon 11 that dramatically reduces full-length mRNA expression of DISC1, but not of shorter forms. We investigated the developmental expression of mouse DISC1 and find that it is highly expressed in the embryonic corpus callosum at a critical time for callosal formation. Taken together our results suggest a significant role for DISC1 in corpus callosum development.

Massive small bowel resection during pregnancy causing short bowel syndrome.

Massive small bowel infarction in pregnancy is rare but has devastating complications. Diagnosis is difficult because pregnancy masks the symptoms. Our aim was to assess risk factors and outcomes of massive resection associated with pregnancy. We conducted a review of nine patients with short bowel syndrome (SBS) secondary to massive bowel resection during pregnancy. The most common cause of bowel resection was midgut volvulus in seven patients. Four of these also had malrotation. Three others had previous abdominal operations, including gastric bypass. The two remaining patients had vascular insufficiency. Five of the nine patients presented after a cesarean delivery. There were three fetal deaths. Resulting small bowel length was less than 60 cm in all but one patient. All patients required parenteral nutrition (PN). Seven patients developed significant complications related to SBS and long-term PN. Four patients underwent transplantation. Massive small bowel resection during pregnancy is a devastating complication, which requires a high degree of suspicion to diagnose. Most patients have risk factors, which include previous surgery, congenital malrotation, and a hypercoagulable state. Surviving patients usually need long-term PN or transplantation.

A 25-year experience with postresection short-bowel syndrome secondary to radiation therapy.

BACKGROUND: Short-bowel syndrome (SBS) can be caused by abdominal and pelvic malignancies treated by radiation therapy (XRT). The management and long-term outcome of these patients is poorly defined. METHODS: This was a retrospective observational study. We reviewed 48 adults developing postresection SBS after XRT over a 25-year period. There were 36 women and 12 men ranging from 19 to 78 years. Follow-up evaluation ranged from 1 to 360 months. RESULTS: The underlying cancer in women included rectum (n = 13), ovary (n = 8), uterus (n = 7), and cervix (n = 6). In men, rectal cancer (n = 4) was most common. The interval to SBS was 1 to 234 months, with 16 (33%) patients developing SBS within 12 months. The indication for surgery was intestinal obstruction in 35, fistula in 9, perforation in 5, and ischemia in 2. Thirty-four (71%) patients underwent multiple resections and residual radiation enteritis was present in 34 (71%). Thirty-six (75%) patients also underwent colectomy and 28 (58%) had an ostomy. Intestinal remnant length was 60 cm or less in 11 patients, 60 to 120 cm in 16 patients, and 120 to 180 cm in 21 patients. Parenteral nutrition was weaned in 9 (19%) patients, and 30 (62%) patients remain on parenteral nutrition. Up to half (48%) of the patients had further intestinal procedures, including 2 liver-small-bowel transplants. Mortality during the follow-up period was 35%, with 8 patients dying within 12 months. CONCLUSIONS: Postresection SBS develops within months to years after XRT for mainly gynecologic and rectal malignancies. Intestinal obstruction is the most common reason for surgery. Multiple resections, colectomy, and ostomy are performed frequently. Long-term survival is possible in many patients although further surgical intervention, including transplantation, can be performed safely.

Permanent diversion rates after neoadjuvant therapy and coloanal anastomosis for rectal cancer.

BACKGROUND: The aim of this study was to assess the rate of permanent diversion in patients undergoing coloanal anastomosis after neoadjuvant therapy for rectal cancer. METHODS: We performed a retrospective review of patients with rectal cancer who underwent a total mesorectal excision of a tumor within 9 cm of the anal verge. RESULTS: There were 201 patients who underwent resection with coloanal anastomosis, with a mean follow-up period of 51 months. The average tumor distance from the anal verge was 7 cm (range, 4-9 cm). Neoadjuvant therapy was administrated in 145 patients, 47 had no radiation, and 9 received radiation postoperatively. Thirty-two patients (16%) had long-term complications including incontinence, fistulas, and strictures. Twenty-five patients (12%) had recurrent disease, 16 of these were local recurrence. The total rate of permanent diversion was 29 (14%). Reasons for diversion included local recurrence in 12 patients (6%), complications in 10 patients (5%), and poor function in 7 patients (3%). CONCLUSIONS: Poor bowel function, late complications, and local recurrence all contribute to permanent diversion after a coloanal anastomosis. Neoadjuvant therapy in conjunction with a total mesorectal excision and coloanal anastomosis leads to acceptably low permanent diversion rates in the vast majority of patients.

Fulminant Clostridium difficile enteritis after proctocolectomy and ileal pouch-anal anastamosis.

Clostridium difficile (C. difficile) infection of the small bowel is very rare. The disease course is more severe than that of C. difficile colitis, and the mortality is high. We present a case of C. difficile enteritis in a patient with ileal pouch-anal anastamosis (IPAA), and review previous case reports in order to better characterize this unusual condition.

Hartmann's colostomy reversal: outcome of patients undergoing surgery with the intention of eliminating fecal diversion.

Reversal of a Hartmann's operation can be a morbid undertaking; successful restoration of intestinal continuity cannot be guaranteed. Between June 2001 and July 2006, 35 Hartmann's reversals were undertaken. There were 19 males (54%). Mean age was 54.7 years (range, 14-82 years). Twenty-one (60%) patients had their Hartmann's for diverticular disease, 7 (20%) for anorectal cancer, 4 (11%) for volvulus, and 3 for miscellaneous reasons. Mean length of stay was 7.7 days (range, 3-16 days); 23 per cent required intensive care for a mean 2.3 days (range, 1-4 days). Blood loss was 470 mL, and mean operative time was 4.28 hours (range, 1-8.3 hours). The mean time interval between the original operation and its reversal was 8.9 months (range, 1.4-55 months). Extensive lysis of adhesions was required in 69 per cent, 40 per cent experienced minor complications (urinary tract infections, ileus, and so on), and 38 per cent had major complications (myocardial infarction, leak, hernias, respiratory failure). There was one death (3%). The operation failed because of intraoperative circumstances in three patients (8%). Ten patients (26%) had stomas at the time of discharge of which 3 were intended to be permanent and 7 were temporary. Of the latter, 3 were successfully closed, 3 are awaiting closure, and 1 had complete anastomotic failure requiring permanent diversion. Total failure rate was 10.3 per cent; contributing factors included prior radiation and ultra-low anastomoses.

Mapping of deletion and translocation breakpoints in 1q44 implicates the serine/threonine kinase AKT3 in postnatal microcephaly and agenesis of the corpus callosum.

Deletions of chromosome 1q42-q44 have been reported in a variety of developmental abnormalities of the brain, including microcephaly (MIC) and agenesis of the corpus callosum (ACC). Here, we describe detailed mapping studies of patients with unbalanced structural rearrangements of distal 1q4. These define a 3.5-Mb critical region extending from RP11-80B9 to RP11-241M7 that we hypothesize contains one or more genes that lead to MIC and ACC when present in only one functional copy. Next, mapping of a balanced reciprocal t(1;13)(q44;q32) translocation in a patient with postnatal MIC and ACC demonstrated a breakpoint within this region that is situated 20 kb upstream of AKT3, a serine-threonine kinase. The murine orthologue Akt3 is required for the developmental regulation of normal brain size and callosal development. Whereas sequencing of AKT3 in a panel of 45 patients with ACC did not demonstrate any pathogenic variations, whole-mount in situ hybridization confirmed expression of Akt3 in the developing central nervous system during mouse embryogenesis. AKT3 represents an excellent candidate for developmental human MIC and ACC, and we suggest that haploinsufficiency causes both postnatal MIC and ACC.