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Notes: Abstract This paper summarizes information on geomorphology and physical resources as a part of the Jordan Badia Research and Development Program. The research focused on the issue of the environment in arid lands as an aid to providing practical options for sustainable development, for the benefit not only of the Hashemite Kingdom of Jordan but of other arid regions of the world. The research is significant in that there is a need to identify usable natural resources and establish a framework for their effective exploitation and management in a marginal, fragile environment, which is sensitive to change. Pressure for development of the Badia stems from the fact that the great majority of the population in Jordan is compressed into less than 10% of the country by environmental constraints. It is hoped that the Jordan Badia Research and Development Program will provide the required framework to ease current environmental pressures, encourage migration to the Badia, a sparsely populated region, and establish economically and ecologically self-supporting communities. This paper discusses the following areas that are related to the sustainable development of the Jordan Badia with special emphasis on the Safawi area in the northern Jordan Badia; geomorphology, including landform, processes, and hazards; geology and physical resources; hydrology; surface water and water engineering; and groundwater.

Notes: Abstract This paper summarizes information on geomorphology and physical resources as a part of the Jordan Badia Research and Development Program. The research focused on the issue of the environment in arid lands as an aid to providing practical options for sustainable development, for the benefit not only of the Hashemite Kingdom of Jordan but of other arid regions of the world. The research is significant in that there is a need to identify usable natural resources and establish a framework for their effective exploitation and management in a marginal, fragile environment, which is sensitive to change. Pressure for development of the Badia stems from the fact that the great majority of the population in Jordan is compressed into less than 10% of the country by environmental constraints. It is hoped that the Jordan Badia Research and Development Program will provide the required framework to ease current environmental pressures, encourage migration to the Badia, a sparsely populated region, and establish economically and ecologically self-supporting communities. This paper discusses the following areas that are related to the sustainable development of the Jordan Badia with special emphasis on the Safawi area in the northern Jordan Badia; geomorphology, including landform, processes, and hazards; geology and physical resources; hydrology; surface water and water engineering; and groundwater.

Notes: Preliminary studies on the proliferative effects of lytic peptides were carried out using NIH 3T3 murine fibroblast cells and human lymphocytes. Cells were cultured in various concentrations of three different amphipathic peptides (SB-37, Shiva-1, and Vishnu), and enhanced proliferation was determined by uptake of 3Hthymidine with treated cells compared with control cultures. Enhanced proliferation of 3T3 cells was observed in cultures containing 50 μM or less SB-37. The primary study consisted of 263 four-cell- to eight-cell-stage mouse embryos from naturally bred mice and incubated in Whitten's medium containing 0.2, 1, or 10 μM of the amino terminus of an amphipathic cecropin B analog (Vishnu) or in Whitten's medium alone. Embryos were cultured to the hatched blastocyst stage, and effect of treatment was determined by the rate of growth to that stage of development. Statistical analysis revealed that culture in all three levels of Vishnu significantly accelerated in vitro growth of these stages of preimplantation embryos compared with controls. These results indicate that Vishnu promotes increased cleavage rates of embryos in vitro. A growth factor receptor clustering mechanism of action is proposed. This peptide may have some potential as an embryo culture medium additive to enhance in vitro growth rate.

Notes: This study was designed to determine what effect electropulse parameters would have on rate of fusion, lysis, and embryo viability when embryos were subjected to electrofusion treatment in nonelectrolyte or electrolyte pulse media. Previous experiments have shown electrolyte medium (i.e., phosphate-buffered saline; PBS) to have a positive effect on electric pulse-induced murine oocyte activation. In addition, these results also indicated that pulse media containing 0.9 mM Ca2+ induced a dramatic increase in the rate of murine oocyte activation compared with oocytes pulsed in media containing 0.0 or 0.05 mM Ca2+. Pronuclear or two-cell-stage embryos were obtained from superovulated prepubertal randomly bred Swiss (albino) female mice. Embryos were randomly assigned to three nonelectrolyte and three electrolyte treatment media. Nonelectrolyte media consisted of 0.3 M mannitol (T1), 0.3 M mannitol + 0.05 mM CaCl2 (T2), and 0.3 M mannitol + 0.9 mM CaCl2 (T3). Electrolyte media consisted of Ca2+ -free PBS (T4), PBS containing 0.05 mM CaCl2 (T5), and PBS containing 0.9 mM CaCl2 (T6). Three experiments were carried out; the objective of the first was to determine the rate of fusion and rate of lysis in murine two-cell embryos placed in the two types of (0.3 M mannitol, T1-T3; and PBS, T4-T6) fusion media and subjected to a fusion procedure (3 V, 5 sec AC alignment pulse, followed by a 1.56 kV · cm-1, 99 μsec DC fusion pulse). Control two-cell embryos were placed in T1 for 2 min and did not receive a fusion pulse. The objective of experiment 2 was to evaluate the development of pronuclear stage embryos to the blastocyst stage in vitro after receiving the fusion pulse in T1, T3, and T6. Control embryos were not subjected to fusion treatment. In experiment 3, T3 and T6 were used to test the rate of fusion and rate of development for pronuclear-stage karyoplasts fused to enucleated pronuclear-stage cytoplasts. Micromanipulations were carried out, and all pronuclear embryos were placed into culture and the number developing to the four-cell stage and subsequently to the blastocyst stage was assessed. Two-cell-stage embryos pulsed in T5 and T6 exhibited significantly higher rates of fusion (96.9 and 92.9%, respectively) compared with T1 (83.7%), T2 (84.7%), or T3 (77.6%) ( P 〉 0.05). There was no significant difference (P 〉 0.05) in rate of lysis between any of the treatment groups. Pronuclear-stage embryos placed in T1, T3, and T6 and subjected to the electrofusion procedure resulted in 55.5%, 52.3%, 50.0%, and 54.2% of T1, T3, T6, and control embryos developing to the blastocyst stage, respectively. There was no difference (P 〉 0.05) between treatment groups in development to the blastocyst stage after 98 hr in culture. Finally, nuclear transplant results indicated no difference in the rate of pronuclear karyoplast-cytoplast fusion between T3 (79.7%) and electrolyte T6 (85.5%) media (P 〉 0.05). There was also no difference in the rate of development to the four-cell stage (78% vs. 72.9%) or blastocyst stage (59.3% vs. 54.2%, T3 and T6, respectively) (P 〉 0.05). However, a difference was observed in rate of development to the four-cell stage and blastocyst stage between nonpulsed control pronuclear stage embryos and T6-treated karyoplast-cytoplast constructs (86.3% vs. 72.9% and 68.5% vs. 54.2%, respectively) (P 〈 0.05). In addition, there was no difference in the rate of lysis observed between T3, T6, and control embryos. These data indicate that the application of a 3 V, 5 sec AC alignment pulse prior to a single 1.56 kV · cm-1 DC fusion pulse to electrolyte PBS results in successful fusion of murine two-cell blastomeres at a rate equal to that of nonelectrolyte 0.3 M mannitol. In vitro development of pronuclear-stage or fused pronuclear transferred karyoplast-cytoplasts after an AC alignment pulse followed by a DC fusion pulse in either 0.3 M mannitol containing 0.9 mM Ca2+ or PB1 does not adversely effect early murine embryonic development in vitro.

Notes: These experiments were designed to test the effects of an electrofusion and an electroporation pulse on bovine sperm-hamster egg development. In experiment 1, single motile sperm were injected into the perivitelline space of each egg. A 4,500 V/cm, 30 μsec fusion pulse (FP) was applied while sperm-egg membrane contact was maintained. It was observed that single motile sperm were rendered immotile immediately after FP application whereas nonpulsed single motile sperm remained motile for up to 36 h postinjection. In addition, both motile and sanicated spermatozoa were injected directly into the ooplasm prior to receiving an FP to determine whether the FP was detrimental to sperm viability. In experiment 2, to induce the acrosome reaction, an 1,150 V/cm electroporation pulse was applied to washed bovine sperm suspended in TALP medium containing 5 mM Ca2+. Treated and nontreated sperm were coincubated with zona-free hamster ova, and sperm-penetrating ability was measured. Results from experiment 1 indicate that FP failed to induce sperm-egg fusion (0/69). FP did not, however, inhibit decondensation or pronuclear formation of sperm injected into hamster egg ooplasm. Single motile sperm injected into the ooplasm resulted in development of both pulsed (19/28) and nonpulsed (21/28) groups. Sonicated tail-free sperm heads injected into the ooplasm resulted in no detectable difference between treated (18/30) and nontreated (19/30) groups. In experiment 2, treatment of sperm with electroporation pulse +5 mM Ca2+ increased zona-free hamster ova penetration scores over nontreated sperm within bulls (P 〈 .05). These results indicate that an electroporation pulse in conjunction with high Ca2+ rather than an electrofusion pulse facilitates sperm penetration and early development.

Notes: These experiments were designed to monitor influx of extracellular Ca2+ into the murine ooplasm following a 1.56 kV · cm-1 direct current (DC) electrofusion pulse and subsequently to determine its effect on rate of activation. Pulse media consisted of nonelectrolyte (0.3 M mannitol) and electrolyte (phosphatebuffered saline; PBS) media each containing 0.0, 0.05, or 0.9 mM Ca2+ (groups T1-T3 and T4-T6, respectively). Cumulus-free oocytes were incubated in 100 μl drops of PBS containing 2 μM of the calcium indicator fluo-3/AM for 60 min at 37°C. Fluo-3/AM-loaded oocytes were equilibrated for 7 min in assigned treatment media (T1-T6) prior to application of DC pulse. Change in fluorescent intensity was monitored for 6.5 min after DC pulse by photon counting spectrofluorometry. Fluorometric measurements demonstrate a dramatic rise in intracellular free Ca2+ (Ca2+i) following DC pulse is associated with Ca2+ ion concentration in the pulse medium. Significantly (P 〈 0.01) higher Ca2+i levels were observed when 0.9 mM Ca2+ was added to the pulse medium (T3 and T6) compared with pulse medium containing lower Ca2+ ion concentrations (T1, T2, T4, and T5; P 〉 0.05). Differences (P 〈 0.01) were observed in peak Ca2+i levels 18 sec after pulse with mean percent change in fluorescence of 5.1%a, 33.9%b, 112.7%c, 1.2%a, 9.3%a, and 99.9%c for T1-T6, respectively (values with different superscripts are significantly different at P 〈 0.01). Increased oocyte membrane permeability to Ca2+ ion after DC pulse was observed for a minimum of 5 min after delivery of the 1.56kV · cm-1 pulse. Nonloaded oocytes receiving the same pulse treatments were cultured in 100 μl drops of Whitten's medium for 8 h prior to evaluation of oocyte activation. Activation was defined as the formation of one pronucleus and one polar body, two or more pronuclei, or two cells each containing a nuclear structure. Activation rates of 28.2%b, 31.1%b, 70.4%d, 35.6%b, 57.3%c, 71.8%d 8.7%a (P 〈 0.01) were observed for T1-T6 and control nonpulsed oocytes, respectively. These data demonstrate that supplementation of 0.9 mM of Ca2+ ion to pulse medium results in a dramatic rise in Ca2+i after DC pulse. This influx of Ca2+ ion has a positive effect on oocyte activation rate.

Notes: An indirect immunofluorescence assay was used to detect the presence of male-specific protein(s) on various stages of preimplantation porcine embryos. Embryos were collected at slaughter from the reproductive tracts of day-2.5, -4, -5, -6, and -8 (day 0 = first day of estrus) sows and gilts. Embryos were placed in medium containing an anti-male primary antibody, washed, and transferred to culture drops containing a fluorescein isothiocyanate (FITC)-labeled secondary antibody. Embryos were classified as either fluorescent (H-Y positive) or nonfluorescent (H-Y negative), transferred to coded drops, and karyotyped to examine sex chromosomes. A total of 91 eight-cell to blastocyst stage embryos were evaluated; of these, 46% were classified as fluorescent and 54% as nonfluorescent. Of readable metaphase spreads (65%) from these embryos, 81% (48 of 59, P 〈 0.005) were correctly sexed by immunological detection of the male-specific antigen. Although 13 % (2/15)of four-cell embryos evaluated were classified as fluorescent, the accuracy with which embryos at this stage were sexed by detection of H-Y antigen was not different from 50%. Fifty percent of eight-cell embryos were classified as H-Y positive with 78% of embryos correctly sexed. It was concluded that the eight-cell embryo is the earliest stage of development for which there is evidence for expression of H-Y antigen. Detection of the male-specific protein was difficult at the expanded blastocyst stage.