Hormone ontogeny in the ovine fetus. XXIII. Pulsatile administration of follicle-stimulating hormone stimulates inhibin production and decreases testosterone synthesis in the ovine fetal gonad

Inhibin, a gonadal glycoprotein with selective FSH-suppressing activity, is synthesized by the Sertoli cell of the testis and the granulosa cell of the ovary mediated by the action of FSH. It is not known whether inhibin is produced by the fetal testes and ovaries or if FSH has the capacity to stimulate inhibin production by the fetal gonad. To explore these questions, we examined the bioactive inhibin content of the gonads of 16 chronically catheterized sheep fetuses between 111 and 143 days gestational age (0.7-0.95 gestation) in an ovine pituitary bioassay. Both the fetal testes and ovary contained inhibin activity (testes, 53.5-1,240 U inhibin/g tissue; ovaries, 58.5-2,250 U/g). After pulsatile administration of oFSH (5 micrograms every 3 h) to the fetus for 5 days in 1 fetus and 10 days in 2 fetuses, 10-day gonadal inhibin content of fetal testes increased to 5,080 +/- 3,180 U/testes (n = 3) vs. 165 +/- 50 U/testes in controls (n = 8; P less than 0.02); the concentration of testicular inhibin in these features rose to a mean of 9,100 +/- 6,620 vs. 415 +/- 126 U/g tissue in controls (P less than 0.01). Ovarian inhibin content in female fetuses given ovine FSH for 10 days was 5,220 +/- 4,920 U/ovary (n = 4) compared to 40 +/- 16 U/ovary in controls (n = 4); the inhibin concentration was 41,000 +/- 30,000 U/g in ovaries of FSH-treated fetuses vs. 1,190 +/- 960 U/g in controls. The ovary of 1 female fetus contained several large follicles and the highest inhibin concentration. Unexpectedly, FSH administration was associated with a decrease in testosterone content in the fetal testes and ovaries. The testosterone content was 0.54 +/- 0.42 ng/ovary after FSH treatment (n = 4) vs. 2.11 +/- 0.68 ng/ovary in controls (n = 4; P less than 0.02). The testosterone concentration fell to 5.8 +/- 2.0 ng/g in treated female fetuses vs. 60.3 +/- 14.6 ng/g in controls (P less than 0.0005). The testosterone content in fetal testes decreased to 21.7 +/- 6.9 ng/testes in FSH-treated fetuses (n = 3) vs. 75.1 +/- 24.0 ng/testes in controls (n = 5; P less than 0.04); the testosterone concentration fell to 38.6 +/- 16.1 ng/g tissue compared to 223.0 +/- 88.7 ng/g in untreated controls (P less than 0.03). In male fetuses the concentration of plasma testosterone decreased to 15.5 +/- 2.3 ng/dl after FSH treatment, significantly lower than 39.6 +/- 4.5 ng/dl in controls (P less than 0.02).(ABSTRACT TRUNCATED AT 400 WORDS)     

Apoprotein synthesis by human duodenojejunal mucosa.

We tested whether human duodenojejunal mucosa is able to synthesize apoproteins from amino acid precursors because lipoprotein-like particles are visualized by electron microscopy in human absorptive cells and because apoproteins are synthesized by the perfused rat intestine. Duodenojejunal biopsies from 21 normal fasting volunteers were incubated with L-[U-14C]leucine; 15.6 +/- 3.3 nmoles of the [14C]leucine was incorporated into protein by 100 mg wet weight of biopsies during an incubation of 2 hr. No [14C]leucine was incorporated by boiled biopsies. Homogenates of incubated biopsies were fractionated by ultracentrifugation: 43 +/- 2% of the incorporated 14C as found in the d less than 1.006 fraction; 3.2 +/- 0.6%, 8.1 +/- 1.0% and 48 +/- 2% were found to be associated with the d = 1.006 to 1.063, d = 1.063 to 1.25, and d greater than 1.25 fractions, respectively. The specific activity in the d less than 1.006 fraction was 5.6 times greater than that in the other fractions. Of the 14C incorporated into the d less than 1.006 fraction, 10.8, 7.2, and 7.1% were specifically precipitated by rabbit antihuman apoproteins A-I, A-II, and B, respectively. Of the 14C incorporated into the d = 1.006 to 1.063 fraction, 11.3 4.1, and 8.5% were specifically precipitated by rabbit antihuman apoprotein A-I, A-II, and B, respectively. Approximately 5% of the 14C incorporated by the d = 1.063 to 1.25 fractions were precipitated by rabbit antihuman apoprotein A-I or A-II. None of the d less than 1.25 fractions precipitated a significant amount of radioactivity with rabit antihuman apoprotein C-II, or antiarginine-rich apoprotein. None of the antibodies precipitated radioactivity from the d greater than 1.25 fraction. These experiments suggest that human duodenojejunal mucosa is able to synthesize in vitro apoproteins A-I, A-II, and B from amino acid precursors. The specificity of the immunoprecipitates was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis.     

Exercise testing at 3 weeks, 6 weeks and 18 months after infarction and the outcome at 3 years in young patients (under 55 years).

To examine the prognostic value of routine postinfarction exercise tests in young patients, exercise tests were carried out at 3 and 6 weeks and 18 months after infarction in 149 patients aged under 55 years at the time of the index infarction. The patients also had coronary angiography and left ventriculography a mean of 3 months after infarction. Three years after infarction, only two of the 149 patients have died, reinfarction occurred in only seven (4.7%) patients; unstable angina in four (3%) patients and coronary artery surgery was needed in 31 (20.8%) patients; 16 in the first, 10 in the second, and 5 in the third year of follow-up. Angina on exercise testing at 6 weeks was the only variable with any predictive value. Eighteen (38%) of the 47 patients with, compared to 12 (11.8%) of the 102 patients without, angina on exercise testing at 6 weeks had coronary surgery (less than 0.001). None of the other exercise variables reliably predicted death, or other complications, including coronary surgery. Ten (13.8%) of the 75 patients excluded from the study died during follow-up; six of them within 6 weeks of infarction. Four (67%) of these patients were excluded from the study because of heart failure. Therefore, the 3-year outcome in young survivors of a myocardial infarction is good and is not reliably predicted by exercise testing at 3 and 6 weeks or 18 months.     

p16INK4A and p15INK4B gene deletions in primary leukemias

The 9p21 locus has been deleted at a high frequency in a wide variety of tumors. Recently, two genes, p16INK4A and p15INK4B (also called MTS1 and MTS2), have been localized in close proximity at the 9p21 locus, encoding cyclin-dependent kinases 4/6 inhibitors of relative molecular mass 16 kD and 15 kD, respectively and also found to be deleted at a high frequency in tumor cell lines. We analyzed p16INK4A and p15INK4B genes in 178 cases of primary leukemias including 81 cases of chronic lymphocytic leukemia (CLL), seven of hairy cell leukemia (HCL), seven of chronic myelogenous leukemia (CML), 43 of acute myelogenous leukemia (AML), 27 of acute lymphoblastic leukemia (ALL), and 13 of myelodysplastic syndrome (MDS) by Southern blot analyses. The ALL cases showed a relatively high frequency of homozygous deletions (22%, 6 of 27) at the p16INK4A gene locus. Interestingly, of the six cases with p16INK4A homozygous deletions, only three showed homozygous deletions at the p15INK4B gene. In 81 CLL patients, we detected one homozygous and five heterozygous deletions at both the p16INK4A and p15INK4B genes and two heterozygous deletions at the p16INK4A gene alone. Deletion of these two genes in AML cases is relatively low (9%). We did not detect deletions in any of the MDS, HCL, and CML cases examined. Sequence analyses of p16INK4A gene of six CLL cases with heterozygous deletion at this locus showed a 27-bp deletion at the splice acceptor site of intron 1 in one case and changes in the coding sequence in three other cases. The data presented in this report showed that (1) p16INK4A and p15INK4B genes are preferentially deleted homozygously in ALL and heterozygously in CLL cases with frequent mutation in the second allele, and (2) p16INK4A gene appears to be more frequently deleted than p15INK4B gene.     

PLP1 gene analysis in 88 patients with leukodystrophy

Pelizaeus–Merzbacher disease (PMD) is caused in most cases by either duplications or point mutations in the PLP1 gene. This disease, a dysmyelinating disorder affecting mainly the central nervous system, has a wide clinical spectrum and its causing mutations act through different molecular mechanisms. Eighty‐eight male patients with leukodystrophy were studied. PLP1 gene analysis was performed by the Multiplex Ligation‐dependent Probe Amplification technique and DNA sequencing, and, in duplicated cases of PLP1, gene dosage was completed by using array‐CGH. We have identified 21 patients with mutations in the PLP1 gene, including duplications, short and large deletions and several point mutations in our cohort. A customized array‐CGH at the Xq22.2 area identified several complex rearrangements within the PLP1 gene region. Mutations found in the PLP1 gene are the cause of PMD in around 20% of the patients in this series.     

Relative Femoral Neck Lengthening Improves Pain and Hip Function in Proximal Femoral Deformities With a High-riding Trochanter

BackgroundComplex proximal femoral deformities, including an elevated greater trochanter, short femoral neck, and aspherical head-neck junction, often result in pain and impaired hip function resulting from intra-/extraarticular impingement. Relative femoral neck lengthening may address these deformities, but mid-term results of this approach have not been widely reported.Questions/purposesDo patients who have undergone relative femoral neck lengthening show (1) less hip pain and greater function; (2) improved radiographic parameters; (3) significant complications requiring subsequent surgery; and (4) progression of osteoarthrosis (OA) or conversion to total hip arthroplasty (THA) at mid-term followup?MethodsWe retrospectively reviewed 40 patients (41 hips) with isolated relative femoral neck lengthening between 1998 and 2006 with sequelae of Legg-Calvé-Perthes disease (38 hips [93%]), slipped capital femoral epiphysis (two hips [5%]), and postseptic arthritis (one hip [2%]). During this time, the general indications for this procedure included a high-riding greater trochanter with a short femoral neck with abductor weakness and symptomatic intra-/extraarticular impingement. Mean patient followup was 8 years (range, 5–13 years), and complete followup was available in 38 patients (39 hips [95%]). We evaluated pain and function with the impingement test, limp, abductor force, Merle d’Aubigné-Postel score, and range of motion. Radiographic parameters included trochanteric height, alpha angle, and progression of OA. Subsequent surgeries, complications, and conversion to THA were summarized.ResultsThe proportion of positive anterior impingement tests decreased from 93% (38 of 41 hips) preoperatively to 49% (17 of 35 hips) at latest followup (p = 0.002); the proportion of limp decreased from 76% (31 of 41 hips) to 9% (three of 35 hips; p < 0.001); the proportion of normal abductor strength increased from 17% (seven of 41 hips) to 91% (32 of 35 hips; p < 0.001); mean Merle d’Aubigné-Postel score increased from 14 ± 1.7 (range, 9–17) to 17 ± 1.5 (range, 13–18; p < 0.001); mean internal rotation increased to 25° ± 15° (range, 0°–60°; p = 0.045), external rotation to 32° ± 14° (range, 5°–70°; p = 0.013), and abduction to 37° ± 13° (range, 10°–50°; p = 0.004). Eighty percent of hips (33 of 41 hips) showed normal trochanteric height; alpha angle improved to 42° ± 10° (range, 27°–90°). Two hips (5%) had subsequent surgeries as a result of lack of containment; four of 41 hips (10%) had complications resulting in reoperation. Fourteen of 35 hips (40%) showed progression of OA; four of 40 hips (10%) converted to THA.ConclusionsRelative femoral neck lengthening in hips with combined intra- and extraarticular impingement results in reduced pain, improved function, and improved radiographic parameters of the proximal femur. Although lack of long-term complications is gratifying, progression of OA was not prevented and remains an area for future research.

Level of Evidence

Level IV, therapeutic study.

Electronic supplementary material

The online version of this article (doi:10.1007/s11999-014-4032-9) contains supplementary material, which is available to authorized users.     

Antiidiotypic IgG crossreactive with Rh alloantibodies in red cell autoimmunity

IgG autoantibodies eluted from RBCs of antiglobulin positive normal blood donors contained at least two antibody populations, an IgG autoantibody (Ab 1), and an IgG population (Ab 2) that agglutinated RBCs coated with some Rh(D) alloantibodies. Eight of 24 autoantibody eluates tested agglutinated 3 of 10 anti-Rh(D) sensitized RBCs. The agglutinating activity was inhibited specifically by preincubation of the autoantibody eluate with the reactive anti-D. The reaction did not require the Fc domain of the anti-Rh(D), since autoantibody eluates agglutinated RBCs coated with F(ab')2 prepared from the reactive anti-D sera. These findings indicate that the RBCs of some antiglobulin- positive blood donors contain an immunoglobulin auto-antiidiotype (Ab 2) against the RBC autoantibody (Ab 1) which is demonstrable through its cross-reactivity with selected Rh(D) alloantibodies. Identification of auto-antiidiotypes in RBC autoimmunity lends support to the idiotype- antiidiotype network hypothesis of immune regulation and is consistent with the bizarre and complex serology of autoimmune hemolytic anemia. The absence of clinical hemolysis in antiglobulin-positive normal blood donors suggests that immunoglobulin idiotype-antiidiotype interactions may play a role in modulating the effects of RBC autoimmunity.     

Effectiveness of methyl-GAG (methylglyoxal-bis[guanylhydrazone]) in patients with advanced malignant lymphoma

We treated 51 patients with advanced malignant lymphoma refractory to conventional therapy with methyl-glyoxal-bis(guanylhydrazone) (methyl- GAG) at doses ranging from 400 to 800 mg/sq m. Therapy was started on a weekly schedule and was switched to every other week in responding patients at the onset of toxicity. Partial responses were observed in 6 of 13 evaluable patients with Hodgkin's disease (46%), 5 of 10 patients with diffuse poorly differentiated lymphocytic lymphoma (50%), 2 of 4 patients with nodular poorly differentiated lymphocytic lymphoma (50%), and 3 of 13 patients with diffuse histiocytic lymphoma (23%). Two of six patients with mycosis fungoides showed objective improvement in cutaneous disease. Toxicity was generally mild and included muscular weakness, myalgia, mucositis, and diarrhea; two patients developed bronchospasm following drug infusions. We conclude that methyl-GAG has major antitumor activity when administered on this schedule to patients with advanced malignant lymphoma. The low degree of toxicity, unique mechanism of action, and minimal myelosuppressive effects suggest that methyl-GAG will prove useful in future trials of combination chemotherapy regimens for the treatment of lymphoma.