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A new approach to the preparation of biologically pure FSH from HMG is described. The method entails selective binding of the LH to antibodies against HCG. The bound LH was separated from FSH by DEAE-C chromatography. An FSH preparation was obtained with a potency of 329.7 IU-FSH/mg and undetectable LH activity at the highest dose tested. This preparation, however, was not homogeneous in the immuno-electrophoretic analysis. The nature of the LH activity »unseparable« from FSH by physico-chemical procedures is discussed.

Background. Arterial traumas of the extremities are quite rare in civilian records; nevertheless, patients with trauma of limbs are admitted daily in emergency departments worldwide. The up-to-date information about epidemiology and treatment (open vs endovascular surgery) comes from war records and it is not always easy getting data on mortality and morbidity in these patients. The aim of this study is to analyse the approach (open or endovascular) and the outcome of patients with vascular trauma of upper limbs (from the subclavian artery) and/or lower limbs (distal to the inguinal ligament), in the greater Milan area. Patients and methods. A retrospective analysis was conducted on data recorded by the emergency departments of two hospitals of the greater Milan between 2009-2017. We collected all patients with arterial injuries of the limbs in terms of demography, injury patterns, clinical status at admission, therapy (open or endovascular approach) and outcomes in terms of limb salvage and survival. Results. We studied 52 patients with vascular trauma of extremities. The main mechanism of trauma was road accident (48.1%), followed by criminal acts (32.7%), self-endangering behaviour (13.5%), work (3.8%) and sport accidents (1.9%). Associated lesions (orthopaedic, neurological and/or venous lesions of the limbs) were present in 39 patients (75%). All patients underwent emergency surgery, forty six patients (88,5%) by open repair (PTFE or greater saphenous vein bypass grafts, arterial suture or ligation) while endovascular approach was used only in 6 patients (11.5%), all treated with embolization. The overall post-operative mortality rate was 5.7% (3 patients). Among survivors, we report 5 major amputations of the lower limbs, 3 of them after bypass graft infection and 2 after graft failure. The rate of limbs salvage was 90.4%. Conclusions Isolated arterial trauma of the extremities are rare, usually they occur in the setting of multiple trauma patients. Despite progresses in surgical techniques, there are still controversies in diagnosis and treatment of these patients. We treated the majority of cases with open surgery (n=46), choosing endovascular approach (embolization performed mainly by interventional radiologists) in difficult anatomic districts. We believe that, during decision making of the surgical strategy, it is important to consider the anatomical site of lesions and the general condition of the patients. Moreover, in case of multiple trauma, we suggest a multidisciplinary approach in order to provide the best medical care to the victims.

Thromboangiitis obliterans, or Buerger disease, is a rare nonatherosclerotic segmental inflammatory vasculitis that generally affects young tobacco smokers. Although no surgical treatment is available, the most effective way to halt the disease's progress is smoking cessation. In this case report, a 29-year-old smoker showed up to emergency department with gangrene of his fifth left toe and extensive plantar ulceration. After investigative angiography, he was diagnosed with Buerger disease. On November 2017, he underwent fifth left toe amputation and hyperbaric therapy. Five months after amputation, the patient was rehospitalized because of surgical wound dehiscence, wide ulceration, and pain. He was treated with lipofilling using the Coleman technique. Two weeks after the fat grafting procedure, the patient suspended pain control medication, and after 2 months, the surgical wound was almost healed. Fat grafting (lipofilling) is mostly used in plastic surgery; it offers regenerative effects, with minimal discomfort for the patient. This case report demonstrates a successful alternative use of lipofilling for this unique condition and opens up new options for use of this technique in other fields.

The further purification of HMG obtained from postmenopausal urine with kaolin-acetone, DEAE-C and permutit chromatography was performed using gel filtration on Sephadex (G 200). The potency of the purest fraction isolated was about 2.5 times higher than that of the starting material. The total recovery of biological activity was 96 %.Several methods using ion-exchange materials for the purification of human pituitary gonadotrophin (HPG) and human menopausal gonadotrophin (HMG) have been proposed.The chromatography of HPG on Sephadex (G 75) and DEAE-Sephadex (A 50) were attempted by Beltendorf et al. (1962).The present paper deals with the further purification, using gel filtration on Sephadex (G 200) of HMG obtained from postmenopausal urine with kaolinacetone, DEAE-cellulose and permutit chromatography. MATERIALS AND METHODSAs starting material for the further purification with gel filtration on Sephadex, HMG Pergonal-26 E43 C was used. This was obtained by Donini el al. (1963) by means of the kaolin-acetone method, followed by 10°/o ammonium acetate in 70% ethanol by DEAE-C purification and, finally, by permutit chromatography.The gel filtration was performed on Sephadex (G 200, 140-400 mesh.). Dry Sephadex

The response of the uterine weight of immature mice to FSH and LH preparations with low contamination by each other was studied. The new approach to the biological assay of LH, which could be designated the »uterine-augmentation« test, was based on injecting intact immature mice with a constant dose (4.44 IU) of a biologically pure FSH preparation plus increasing amounts of extracts containing LH. A dose as little as 0.068 IU of LH is capable to increase the uterine weight when injected together with 4.44 IU of biologically pure FSH. A significant uterine response can be obtained when animals are injected with 29.4 μg of 2nd IRP-HMG (equivalent to 0.128 IU-LH and 0.128 IU-FSH). When a constant dose of 4.44 IU of biologically pure FSH is added to each dose level of the 2nd IRP-HMG, a significant uterine response is obtained with 14.7 μg of this reference preparation. The end point was the mouse uterine weight. Some determinations of the LH activity in urinary extracts have been performed according to this new method and from the results it seems that the LH activities, as determined by the ovarian ascorbic acid depletion method, agree with those determined by the uterine-augmentation test. It also seems that the uterine weight response is proportional to the LH activity and not dependent on the FSH-LH ratio of the samples assayed.

Results of this study indicate that some methods for purification and separation of urinary FSH and LH are available and that different grades of separation of the two hormones can be achieved depending on the purity of the starting material used. The most potent FSH fraction with the highest FSH/LH ratio was obtained with chromatography on DEAE-C column, followed twice by gel filtration on Sephadex G-100. Nevertheless all the FSH fractions reported are more or less contaminated by luteinizing hormone.

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