Characterization and Expression of Glutamate Dehydrogenase in Response to Acute Salinity Stress in the Chinese Mitten Crab, Eriocheir sinensis

Glutamate dehydrogenase (GDH) is a key enzyme for the synthesis and catabolism of glutamic acid, proline and alanine, which are important osmolytes in aquatic animals. However, the response of GDH gene expression to salinity alterations has not yet been determined in macro-crustacean species.GDH cDNA was isolated from Eriocheir sinensis. Then, GDH gene expression was analyzed in different tissues from normal crabs and the muscle of crabs following transfer from freshwater (control) directly to water with salinities of 16‰ and 30‰, respectively. Full-length GDH cDNA is 2,349 bp, consisting of a 76 bp 5'- untranslated region, a 1,695 bp open reading frame encoding 564 amino acids and a 578 bp 3'- untranslated region. E. sinensis GDH showed 64-90% identity with protein sequences of mammalian and crustacean species. Muscle was the dominant expression source among all tissues tested. Compared with the control, GDH expression significantly increased at 6 h in crabs transferred to 16‰ and 30‰ salinity, and GDH expression peaked at 48 h and 12 h, respectively, with levels approximately 7.9 and 8.5 fold higher than the control. The free amino acid (FAA) changes in muscle, under acute salinity stress (16‰ and 30‰ salinities), correlated with GDH expression levels. Total FAA content in the muscle, which was based on specific changes in arginine, proline, glycine, alanine, taurine, serine and glutamic acid, tended to increase in crabs following transfer to salt water. Among these, arginine, proline and alanine increased significantly during salinity acclimation and accounted for the highest proportion of total FAA.E. sinensis GDH is a conserved protein that serves important functions in controlling osmoregulation. We observed that higher GDH expression after ambient salinity increase led to higher FAA metabolism, especially the synthesis of glutamic acid, which increased the synthesis of proline and alanine to meet the demand of osmoregulation at hyperosmotic conditions

Dual-drug co-crystal synthesis for synergistic in vitro effect of three key first-line antiretroviral drugs

Two pharmaceutical solid state co-crystals EFZA and NEF of two non-nucleoside reverse transcriptase inhibitors (efavirenz, EFV and nevirapine, NVP) and a nucleoside reverse transcriptase inhibitor, zidovudine (AZT) were prepared. Currently, these drugs are essential in various front-line HIV treatment regimens due to their potential in the management of HIV infection. Solid-state principles and solvent-assisted grinding were used for screening of co-crystal preparation. Characterization was carried out using DSC, HSM and TGA, then complimented by FT-IR spectral data and powder X-ray diffraction. HPLC analysis was used for a solubility assays. Both co-crystals indicated different thermal behaviour compared to the individual drug components. Differences in vibrational modes within the spectral region and the unique powder X-ray patterns confirmed the identity of EFZA and NEF as new molecular derivatives. Quantitative studies using HPLC indicated an enhanced solubility of the co-crystals at different pH values compared to the parent compounds; thus making these interesting candidates for future formulation6

700 Laparoscopic Pancreatic Resections in a French Center: Evolution of the Indications and Outcome

Introduction: Large monocentric series of laparoscopic pancreatic resection (LPR) are still lacking. This series studied evolution of indications and outcome of LPR. Methods: This is a retrospective monocentric study. Main contraindications were major vascular invasion and pancreatitis. Demographics, surgical, and postoperative outcome data were studied. We compared the first 350 LPR (2008-2016) and the last 350 LPR (2017-2020). Results: From 2008 to 2020, 700 LPR were performed including 191 PD (27%), 344 DP (50%), 92 CP (13%), 69 Enucleation (10%) and 4 TP (1%). The applicability of laparoscopy increased from 10% before 2010 to 40% in 2020. Mean age and BMI were 56 (17-87) and 25 kg/m2 (15-48), respectively and 406 were female (58%).The main indications: IPMN (169; 24%), NET (167; 24%) and pancreatic adenocarcinoma (124, 18%). The surgical outcomes showed conversion (3%), mean operative duration (207; 30-540 mn), blood loss (213; 0-2500 ml), transfusion (4%), mortality (9; 1.3%), overall morbidity (56%), POPF B/C (19%), bleeding (7%), re-intervention (7%), readmission (6%) and mean hospital stay 16(2-104). The second period versus the first period showed more comorbidities (39% vs 52%; p<0.0001), less noninvasive IPMN (39% vs 18%; p<0.001, less splenectomy (17% vs 9%,p=0.002), less conversion (4% vs 2%,p=0.046), less POPF (24% vs 15%,p=0.001), less bleeding (9% vs 5%,p=0.036), less re-intervention (10% vs 5%,p=0.004) and a shorter hospital stay (18 vs 13; p<0.001). Conclusion: This large series including all types of pancreatic resections showed more patients with comorbidities and evolution in the indications. The good outcome is furthermore improved with the experience

Indocyanin Green (ICG) for Colonic Viability Assessment in Left Pancreatic Minimal Invasive Surgery (MIS)

Background: Large distal pancreatic tumors may involve the transverse or splenic flexure mesocolon. R0 resection of such lesions may be extended to the mesocolon. Interruption of the arc of Riolan may be difficult to assess especially in MIS. Case report: We report a case of a 17 years old male presenting epigastric pain. The CT scan showed two lesions of the distal pancreas of 7x8cm and 4x3.5cm with aspects compatible with a solid pseudopapillary tumor. The octreo PET showed no fixation of the two lesions. A laparoscopic distal pancreatectomy with splenic preservation was attempted. Transverse and splenic flexure mesocolon were involved by the lesions. The dissection of the mesocolon to release the tumors lead to probable interruption of arc of Riolan and to a doubt on colonic optimal vascularisation after the surgery. We therefore used ICG to assess the viability of the colonic splenic flexure. Discussion and Conclusion: This short video illustrates how ICG can be simply used in left pancreatic surgery when R0 resection requires removal of splenic flexure mesocolon and partial interruption of arc of Riolan. In this case, ICG confirms the adequate perfusion of the splenic flexure and its safe preservation without any need of colonic resection

Laparoscopic Pancreatoduodenectomy with Venous Resection Is Safe Compared to Open Pancreatoduodenectomy: A Monocentric Comparative Study

Background: Laparoscopic pancreatoduodenectomy (LPD) for pancreatic ductal adenocarcinoma (PDAC) is more frequently performed and few small series report on its feasibility with venous resection. In this study, we compared the outcomes of LPD and open PD (OPD) with venous resection. Method: In the study centre and since 2014, LPD with venous resection is performed in selected patients. We retrospectively analysed data from patients who underwent LPD or OPD for PDAC and venous resection between 2014 and 2022. In OPD group, we selected patients who could theoretically benefit from the laparoscopic approach. Results: Eighty nine patients were included, 21 (24%) LPD and 68 (76%) OPD. LPD compared to OPD showed similar median age (69 years and 67, p= 0.3), proportion of female patients (52% and 50%, p= 0.3), median body mass index (24 for both, p= 1.00) and less neoadjuvant chemoradiotherapy (43% versus 79%, p= 0.002). LPD had more lateral venous resections (90% vs 63%, p= 0.028), more reconstruction with autologous peritoneal patch (57% versus 19%, p= 0.002), longer median clamping time (28 vs 15 minutes, p< 0.001) and operative time (360 vs 255 minutes, p< 0.001). There was 1 (5%) venous reconstruction thrombosis in LPD patients and 6 (9%) in OPD patients with no difference (p= 1.00). The outcome showed 1 (5%) death in the OPD group and similar Clavien-Dindo ≥ 3 complications (LPD 14% vs OPD 16%, p= 0.236) and median hospital stay (14 days, p= 0.804). LPD and OPD showed similar T stage [T0 (5% vs 6%), T1/2 (62% vs 82%), T3 (33% vs 12%), p=0.336], median number of lymph nodes harvested (22 vs 23, p=0.893) and invaded (0, p= 0.306), R0 resection rate (81% vs 89%, p=0.286) and adjuvant chemotherapy (90% vs 78%, p=0.342). Conclusion: LPD with venous reconstruction for PDAC is feasible in selected patients and high volume centres with security profile and oncologic outcomes similar to OP

Retromesenteric Omental Flap as Arterial Coverage in Pancreaticoduodenectomy: A Novel Technique to Prevent Post-pancreatectomy Hemorrhage

Background: Post-pancreatectomy hemorrhage (PPH) occurs in 10-15% of patients following pancreaticoduodenectomy (PD), mainly in association with clinically relevant postoperative pancreatic fistula (CR-POPF). Prevention of PPH by arterial coverage (AC) with a round ligament plasty or an omental flap is controversial. This study evaluated clinically relevant PPH (CR-PPH) rate following AC with an original retromesenteric omental flap (ROF) in patients undergoing PD. Methods: This single center retrospective study included 812 open PD (2012-2021) and compared 146 procedures with AC using a ROF (AC-ROF) to 666 PD without AC (no AC). The Fistula Risk Score (FRS; Callery et al., J Am Coll Surg 2013) was calculated. CR-POPF and CR-PPH were recorded according to the ISGPS classification. Results: There were more patients with a high FRS (7-10) in the AC-ROF group: 18 (12%) vs. 48 (7%) with no AC (p<0.01). CR-POPF ocurred in 28 (19%) in the AC-ROF group versus 165 (25%), respectively (p=0.001). CR-PPH was less frequent in the AC-ROF group 5 (3%) versus 66 (10%) with no AC, respectively (p=0.01). There was no reoperations for PPH or POPF in the AC-ROF versus 25 (4%) with no AC (p=0.02). In the 65 patients (8% of the overall population) with FRS=7-10, the CR-PPH rate was 1/20 (5%) in the AC-ROF group vs. 7/45 (16%) in the no AC groupé (p=0.23). Conclusions: AC with ROF can reduce rates of CR-PPH following PD especially in patients at high risk for POPF. This simple and cost-free technique should be further evaluated to confirm these results

Distal Pancreatectomy for Body-pancreatic Ductal Adenocarcinoma: Is Splenectomy Necessary?

Purpose: The value of splenectomy during distal pancreatectomy (DP) for body-pancreatic ductal adenocarcinoma (B-PDAC) has not been evaluated. This study aimed to assess the impact of spleen preservation (SP+) on morbidity and oncological outcomes following DP for B-PDAC. Method: Single-center cohort study including patients who underwent DP with SP+ according to Warshaw’s technique and DP with splenectomy (SP-) for B-PDAC from 2008 to 2019. Exclusion criteria were: distance <5cm between tumor and spleen hilum and multi-organ resection. Primary endpoints were disease-free (DFS) and overall survival (OS). Secondary endpoints were 90-day morbi-mortality using comprehensive complication index (CCI) and ISGPS definitions. Results: 129 patients were enrolled including 59 (46%) SP+ and 70 (54%) with splenectomy (SP-). Median age and BMI were 68 years and 24 kg/m2. SP+ and SP- patients were comparable regarding preoperative variables including neoadjuvant treatment (overall=24%), and laparoscopic approach (overall=39%). There was no 90-day mortality. Hospital stay was shorter after SP+ (11 vs. 16 days; p<0.001). SP+ patients experienced a lower CCI (8.7 vs. 16.6; p=0.004) with lower rates of grade B/C POPF (14% vs. 29%; p= 0.039) and deep organ space abscess (5% vs. 19%; p=0.041). The rate of R0 margins was similar in SP+ and SP- patients (75% vs. 71%, respectively; p=0.840), as well as invaded/harvested node ratio (0.0% vs. 0.0%; p=0.246), and AJCC staging (p=0.200). After a median follow-up of 63 months (IC95%:52-96), 1- ,3- and 5-year DFS and OS were 77%, 50% and 43% and 91%, 63% and 47%, respectively. On multivariate analysis, after doubly-robust adjustments on preoperative data, SP+ was associated with a better DFS (HR=0.315 [IC95%: 0.146-0.678]; p=0.0032), and had no influence on OS (HR=0.664 [IC95%: 0.317-1.392]; p=0.2782). After doubly-robust adjustments on pathological data and perioperative treatment, SP+ had no negative impact on DFS (HR=0.58 [IC95%: 0.3-1.13]; p=0.111) or OS (HR=0.9 [IC95%: 0.47-1.71]; p=0.738). Conclusion: SP+ DP for B-PDAC is associated with less postoperative morbidity than splenectomy, without impairing oncological outcomes. This study provides a rational to evaluate SP+ DP as a potential new oncological standard in B-PDAC