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The hair follicle (HF) is an ideal system for studying the biology and regulation of adult stem cells (SCs). This dynamic mini organ is replenished by distinct pools of SCs, which are located in the permanent portion of the HF, a region known as the bulge. These multipotent bulge SCs were initially identified as slow cycling label retaining cells; however, their isolation has been made feasible after identification of specific cell markers, such as CD34 and keratin 15 (K15). Here, we describe a robust method for isolating bulge SCs and epidermal keratinocytes from mouse HFs utilizing fluorescence activated cell-sorting (FACS) technology. Isolated hair follicle SCs (HFSCs) can be utilized in various in vivo grafting models and are a valuable in vitro model for studying the mechanisms that govern multipotency, quiescence and activation.
Developmental Biology, Issue 110, Stem cells, hair follicle, mouse, dorsal skin dissection, cell sorting, FACS
Isolating Hair Follicle Stem Cells and Epidermal Keratinocytes from Dorsal Mouse Skin
Despina Soteriou, 1 Lana Kostic, 1 Egor Sedov, 1 Yahav Yosefzon, 1 Hermann Steller, 2 and Yaron Fuchs 1
In Sweden, a particular subtype of verocytotoxin-producing Escherichia coli (VTEC) O157:H7, originally defined as being of phage type 4, and carrying two vtx2 genes, has been found to cause the majority of reported human infections during the past 15 years, including both sporadic cases and outbreaks. One plausible explanation for this could be that this particular subtype is better adapted to colonise cattle, and thereby may be excreted in greater concentrations and for longer periods than other VTEC O157:H7 subtypes.
In an experimental study, 4 calves were inoculated with 109 colony forming units (cfu) of strain CCUG 53931, representative of the subtype VTEC O157:H7 (PT4;vtx2;vtx2c). Two un-inoculated calves were co-housed with the inoculated calves. Initially, the VTEC O157:H7 strain had been isolated from a dairy herd with naturally occurring infection and the farm had previously also been linked to human infection with the same strain. Faecal samples were collected over up to a 2-month period and analysed for VTEC O157 by immuno-magnetic separation (IMS), and IMS positive samples were further analysed by direct plating to elucidate the shedding pattern. Samples were also collected from the pharynx.
All inoculated calves proved culture-positive in faeces within 24 hours after inoculation and the un-inoculated calves similarly on days 1 and 3 post-inoculation. One calf was persistently culture-positive for 43 days; in the remainder, the VTEC O157:H7 count in faeces decreased over the first 2 weeks. All pharyngeal samples were culture-negative for VTEC O157:H7.
This study contributes with information concerning the dynamics of a specific subtype of VTEC O157:H7 colonisation in dairy calves. This subtype, VTEC O157:H7 (PT4;vtx2;vtx2c), is frequently isolated from Swedish cattle and has also been found to cause the majority of reported human infections in Sweden during the past 15 years. In most calves, inoculated with a representative strain of this specific subtype, the numbers of shed bacteria declined over the first two weeks. One calf could possibly be classified as a high-shedder, excreting high levels of the bacterium for a prolonged period.
Experimental infection in calves with a specific subtype of verocytotoxin-producing Escherichia coli O157:H7 of bovine origin
Malin E Jonsson,corresponding author1 Erik Eriksson,corresponding author2 Sofia Boqvist,2,3 Anne Margrete Urdahl,1 and Anna Aspan2
AIM: To compare laparoscopic pancreaticoduodenectomy (TLPD) during the initial learning curve with open pancreaticoduodenectomy in terms of outcome and costs.
METHODS: This is a retrospective review of the consecutive patients who underwent TLPD between December 2009 and April 2014 at our institution. The experiences of the initial 15 consecutive TLPD cases, considered as the initial learning curve of each surgeon, were compared with the same number of consecutive laparotomy cases with the same spectrum of diseases in terms of outcome and costs. Laparoscopic patients with conversion to open surgery were excluded. Preoperative demographic and comorbidity data were obtained. Postoperative data on intestinal movement, pain score, mortality, complications, and costs were obtained for analysis. Complications related to surgery included pneumonia, intra-abdominal abscess, postpancreatectomy hemorrhage, biliary leak, pancreatic fistula, delayed gastric emptying, and multiple organ dysfunction syndrome. The total costs consisted of cost of surgery, anesthesia, and admission examination.
RESULTS: A total of 60 patients, including 30 consecutive laparoscopic cases and 30 consecutive open cases, were enrolled for review. Demographic and comorbidity characteristics of the two groups were similar. TLPD required a significantly longer operative time (513.17 ± 56.13 min vs 371.67 ± 85.53 min, P < 0.001). The TLPD group had significantly fewer mean numbers of days until bowel sounds returned (2.03 ± 0.55 d vs 3.83 ± 0.59 d, P < 0.001) and exhaustion (4.17 ± 0.75 d vs 5.37 ± 0.81 d, P < 0.001). The mean visual analogue score on postoperative day 4 was less in the TLPD group (3.5 ± 9.7 vs 4.47 ± 1.11, P < 0.05). No differences in surgery-related morbidities and mortality were observed between the two groups. Patients in the TLPD group recovered more quickly and required a shorter hospital stay after surgery (9.97 ± 3.74 d vs 11.87 ± 4.72 d, P < 0.05). A significant difference in the total cost was found between the two groups (TLPD 81317.43 ± 2027.60 RMB vs laparotomy 78433.23 ± 5788.12 RMB, P < 0.05). TLPD had a statistically higher cost for both surgery (24732.13 ± 929.28 RMB vs 19317.53 ± 795.94 RMB, P < 0.001) and anesthesia (6192.37 ± 272.77 RMB vs 5184.10 ± 146.93 RMB, P < 0.001), but a reduced cost for admission examination (50392.93 ± 1761.22 RMB vs 53931.60 ± 5556.94 RMB, P < 0.05). CONCLUSION: TLPD is safe when performed by experienced pancreatobiliary surgeons during the initial learning curve, but has a higher cost than open pancreaticoduodenectomy.
Cost, Initial learning curve, Laparoscopic surgery, Pancreaticoduodenectomy, Postoperative event
Outcome and costs of laparoscopic pancreaticoduodenectomy during the initial learning curve vs laparotomy
Chun-Lu Tan, Hao Zhang, Bing Peng, and Ke-Zhou Li
2015 May 7